MSA 2012, Yale

MSA 2012 Abstracts

Contributed Oral Session 10
Day: Tuesday, 17 July
Location:
Time: 1715

: Host-parasite relationships between California boletes and bolete-infecting Hypomyces species.

Gilberto Uribe Valdez* and Greg W Douhan. Department of Plant Pathology and Microbiology, University of California, Riverside, CA 92521, USA.

Fungi in the genus Hypomyces are mycoparasites of various fungal groups, including boletes. Until recently, most studies within this genus primarily dealt with taxonomic descriptions without much focus on their biology in natural ecosystems. Currently, there are eight formally described bolete-infecting Sepedonium anamorphs linked to the Hypomyces teleomorph. Our current objective is to determine the level of host specificity between these parasites and their hosts using a phylogenetic approach, since previous reports regarding specificity have been based mostly on morphological observations and a small molecular dataset. Our long-term goal is to then determine whether we can infer any patterns of co-evolution within this system. As determined in a previous study in California, two cryptic species were found within H. microspermus and H. chrysospermus sensu lato, though based on a very limited sample size. We have set out to corroborate the presence of these multiple, phylogenetic species with a multi-gene phylogeny approach. Currently, sequencing of the ITS regions, actin, and elongation factor genes support the multispecies complex hypothesis, though more sequencing will be done to further strengthen our findings. Also, two additional species previously reported to occur in California, H. ampullosporum and H. laevigatum, will be further analyzed and included in our new host-parasite analyses. Currently, we have collected over 400 parasitized boletes within California, cultured the parasites, and isolated DNA from both the hosts and pathogens. Putative basidiomycete-specific primers are being used to identify and establish host relationships based on sequencing of the variable ITS regions and a partial region of the more conserved LSU-rDNA. These host sequences are being used to create species level phylogenies that will be compared to phylogenies of the parasite. We are finding that many bolete clades are being parasitized by closely related Hypomyces isolates, and results for all samples in our collection will be presented.

Contributed Oral Session 13
Day: Wednesday, 18 July
Location:
Time: 0845

: Mating-type heterokaryosis and population shifts in Aspergillus flavus

Rodrigo A Olarte1*, Bruce W Horn2, Carolyn J Worthington1, and Ignazio Carbone1. 1Department of Plant Pathology, North Carolina State University, Raleigh, NC 27695, 2National Peanut Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, Dawson, GA 39842.

Aspergillus flavus is a fungal pathogen of many agronomically important crops worldwide. We sampled A. flavus strains from a cornfield in Rocky Mount, NC. This field was planted in 2010 and plots were inoculated at tasseling with either AF36 or NRRL 21882 (=Afla-Guard) biocontrol strains, both of which are mating type MAT1-2. Subsequently, toxigenic strain NRRL 3357 (MAT1-1) was applied to all plots, including control plots not inoculated with biocontrol strains. Sclerotia were harvested from infected corn ears approximately 4.5 months after planting (2.5 months after biocontrol treatment); ninety single-ascospore isolates were isolated from ascocarps originating from plots treated with AF36 and NRRL 21882. In addition, eighty A. flavus isolates were collected from soil one month after planting (before biocontrol application) and one year after biocontrol application, for a grand-total of 250 isolates. Aflatoxin (AF) and cyclopiazonic acid (CPA) production were determined using standard thin-layer chromatography and HPLC. Three distinct toxin classes were identified: AF-/CPA-, AF+/CPA+ and AF-/CPA+. PCR amplification revealed grouping of isolates into three distinct mating-type classes: MAT1-1, MAT1-2 and MAT1-1/MAT1-2. A significant proportion (54%) of isolates sampled prior to biocontrol treatments were heterokaryotic for mating type (MAT1-1/MAT1-2), and 39% of isolates obtained from ascospores were heterokaryotic as well as 9% of isolates from soil after biocontrol treatments. The vertical transmission of MAT1-1/MAT1-2 to progeny ascospore isolates suggests that heterokaryosis can be maintained in subsequent generations. The population genetic structure before and after the application of biocontrol treatments will be discussed. Further characterization of heterokaryons and their frequency in A. flavus populations may be important in understanding the adaptation of these fungi to changing environmental conditions.

Contributed Oral Session 16
Day: Wednesday, 18 July
Location:
Time: 1115

: Influences of nitrogen deposition and soil warming on saprophytic fungal community structure, fungal growth, and litter decomposition

Christopher M Sthultz1*, Linda TA Van Diepen2, Serita D Frey2, and Anne Pringle1. 1Organismic & Evolutionary Biology Harvard University 16 Divinity Avenue Cambridge, MA 02138, 2Department of Natural Resources and the Environment University of New Hampshire 56 College Road James Hall Durham, NH 03801.

Impacts of global change on microbial communities remain poorly understood. However, because microbes are important drivers of biogeochemical cycles, anthropogenically mediated changes may have strong influences on ecosystem function. Saprophytic fungi are primarily responsible for decomposition in temperate forest systems, yet relatively little is known about whether global changes, including nitrogen deposition and soil warming, will alter community assembly processes. Here we focus on results from the first year of a multi-year litterbag/decomposition experiment using long-term experimental N addition (control, low and high N treatments) and soil warming plots (control and elevated 5oC treatments) in a northeastern hardwood forest. We examined saprophytic fungal community structure, fungal growth, and decomposition from 95 litter bags harvested after 1 year in the field. Using metagenomics, culturing, and laboratory microcosms we tested the influence of nitrogen deposition and soil warming on fungal communities and ecosystem function. We document four major patterns: 1) Fungal communities in elevated nitrogen deposition and soil warming plots were different from control plots. 2) Decomposition varied between treatment and control plots in both experiments. 3) In the nitrogen experiment fungal community composition and decomposition were influenced not only by treatment but also by the source of the litter in the bags (from control, low, or high N plots). There was a significant interaction effect on the community composition. 4) There were differences in growth rates of the same fungal species cultured from each of the three nitrogen treatments. Our results increase understanding of the ecology and evolution of saprophytic fungi in a global change context, and add to what is known about the biodiversity of decomposer fungi. In addition, our results suggest global change may influence the fungi found in the environment, but also the evolution of some fungal species.

Contributed Oral Session 16
Day: Wednesday, 18 July
Location:
Time: 1045

: Diversity and composition of soil fungal communities under long-term nitrogen enrichment

Eric W Morrison1*, Serita D Frey2, W Kelley Thomas3, and Anne Pringle4. 1Department of Molecular, Cellular and Biomedical Sciences, University of New Hampshire, Durham, NH, 2Department of Natural Resources and the Environment, University of New Hampshire, Durham, NH, 3Hubbard Center for Genome Studies, University of New Hampshire, Durham, NH, 4Organismic and Evolutionary Biology, Harvard University, Cambridge, MA.

Nitrogen (N) deposition from fossil fuel burning has the potential to affect ecosystem processes such as the decomposition and storage of soil organic matter. The Harvard Forest Chronic Nitrogen Addition experiment (HFCN) was established in 1989 to test the effects of long-term N fertilization on ecosystem processes in a northeastern mixed-hardwood forest. Three plots receive one of three treatments: ambient N deposition (control), 50 kg N ha -1 yr -1 (low N), or 150 kg N ha -1 yr -1 (high N). Researchers at this site have observed an accumulation of soil C in the N fertilized plots and a decrease in fungal biomass, ligninolytic enzyme activity, and rates of litter decay. Soil fungi are the primary decomposers of lignin in these communities. We hypothesized that decreased decomposition rates in N fertilized plots may be due to decreased diversity and changes in the composition of the fungal community. We performed a marker gene study of the fungal community in the organic soil horizon using 454 sequencing of three loci: ITS1, ITS2, and rDNA large subunit D2-D3 region. The dominant fungal family in soils under ambient N deposition was the Russulaceae. This family underwent a significant decrease in relative abundance in N treated soils. Unknown fungi dominated N treated soils. Control soils had significantly fewer unknown OTUs. High N soils had higher numbers of OTUs and singleton OTUs then control soils and low N soils, and had higher predicted richness. Fungal communities in high N soils had different community structure than control and low N soils as predicted with OTU based and phylogenetic beta-diversity metrics. Differences in community composition and higher numbers of unknown OTUs in high N soil fungal communities may suggest that a previously uncatalogued portion of the community is released by the decline of the dominant Russulaceae.

Contributed Oral Session 14
Day: Wednesday, 18 July
Location:
Time: 0845

: Surveys at five sites across North America reveal taxonomic and functional differences among fungi from living, senescent, and fallen leaves

Jana M U'Ren* and A Elizabeth Arnold. University of Arizona, School of Plant Sciences, Tucson, AZ 85719.

Some fungi recovered from the interior of living, asymptomatic leaves (endophytes) have been isolated from leaf litter, suggesting that many of these symbiotrophs are decomposer fungi with only a transiently endophytic life stage. However, communities rarely are sampled to statistical completion and surveys rarely encompass multiple host species and sites. We examined the specificity, composition, and geographic turnover of fungi inhabiting the interior of leaves of the same host individuals and in the same seasonal cohorts but that represented three different stages (living, senescent/dead attached to branches, fallen/decomposing). Analysis of 2064 isolates from 3-4 abundant species of woody plants in five temperate and boreal sites consistently revealed that fungal communities change in response to leaf senescence and incorporation into leaf litter. Although boundaries among leaf life-stages were blurred by some fungi that occurred in both living and dead leaves, genotypes that were highly abundant in leaf litter were seldom found as endophytes. Communities in each leaf stage differed among hosts and sites, but fungi in dead leaves were less host- and geographically specific than those in living leaves. Fungi from each leaf class differed in their relationships with environmental factors, with fungi in dead leaves demonstrating less strict association with precipitation and growing season duration than endophytes. On-going work pairing these data with functional traits will inform the breadth of ecological roles of endophytes and provide the basis for hypothesis testing with culture-free methods.

Symposium 3 - The mycologist's guide to the new International Code of Nomenclature for algae, fungi, and plants
Day: Monday, 16 July
Location:
Time: 1740

: Subcommissions, working groups, and protected lists

Keith A Seifert1* and Andrew N Miller2. 1Eastern Cereal & Oilseed Research Centre, Agriculture & Agri-Food Canada, Ottawa, Ontario, Canada K1A 0C6, 2University of Illinois, Illinois Natural History Survey, Champaign, IL 61820-6970 USA.

The International Commission on the Taxonomy of Fungi (ICTF) is a 20-member group affiliated with both the International Mycological Association and the International Union of Microbiological Societies. Its activities over the past 30 years have been focused on the taxonomy and nomenclature of economically important fungal groups, with subcommissions or affiliated commissions working on Penicillium and Aspergillus, Trichoderma and Hypocrea, and Fusarium. Informal working groups on Ceratocystis and Ophiostoma, and on the Mycosphaerella complex, have been periodically active. In the wake of the nomenclature changes discussed in this symposium, additional groups on Colletotrichum and the Hypocreales are in the process of forming. The activities of these groups will briefly be reviewed in this talk. Although over the long term the ICTF will continue its focus on economically important fungi, in the short term we plan to assist with the formation of working groups to address the broadest possible taxonomic scope. This is an exciting time for increased cooperation and collaboration among fungal taxonomists, as we work towards developing consensus on the selection of genus and species names, and the development of lists of protected and/or rejected names.

Contributed Oral Session 1
Day: Monday, 16 July
Location:
Time: 0930

: Identification of fungal contamination in bottled maple syrup

Seanna L Annis1*, Rafael Garcia2, Beth Calder2, and Kathryn L Hopkins3. 1School of Biology and Ecology, University of Maine, Orono, ME, 04469, 2Department of Food Science and Human Nutrition, University of Maine, Orono, ME, 04469, 3Cooperative Extension, University of Maine, Orono, ME, 04469.

Maple syrup processors in the northeastern USA occasionally observe fungal contamination in their bottled maple syrup containers. The concern is that the contaminants may pose a health risk. We identified fungal organisms in 32 bottles of syrup from different processors in the northeastern USA. Most containers had one fungus, but some were contaminated with multiple fungi. Fungi were identified by morphology and DNA sequences of their ribosomal internal transcribed spacer regions and B tubulin genes. Multiple species of the genera Penicillium and Aspergillus, single species of Wallemia, and as yet unidentified yeasts were isolated from multiple bottles. Species of Paecolimyces and Cladosporium were isolated from single bottles of syrup. Some of the genera of Penicillium and Aspergillus identified are known to produce mycotoxins, and the production of these compounds in maple syrup is being evaluated. Syrup is typically bottled at 82 C to decrease the risk of microbial contamination. Spores from some of the fungi were able to germinate after treatment at 70 C for 3 minutes. A higher bottling temperature may be one of the changes in bottling practices required to minimize fungal contamination. This research will result in new recommendations to maple syrup processors on preventing future fungal contamination of bottled syrup.

Contributed Oral Session 1
Day: Monday, 16 July
Location:
Time: 1000

: Marine ascomycetes of U.S. Gulf of Mexico barrier island beaches: biodiversity, succession and sampling strategy

Allison K Walker. The University of Southern Mississippi, Gulf Coast Research Laboratory, 703 East Beach Dr, Ocean Springs, MS 39564.

Saprotrophic marine ascomycetes are an important but often overlooked component of marine ecosystems, vital to coastal nutrient cycling processes and food webs. The existence of a unique marine mycota was discovered only within the past 50 years and basic marine fungal distribution data are lacking in many parts of the world. The sampling intensity required to characterize the biodiversity of marine fungal communities is unknown, despite the fact that marine and estuarine environments occupy 75% of the globe. Previous studies have shown that subtropical zones contain temperate, cosmopolitan, sub-tropical and tropical marine fungal species, and thus are potentially highly diverse. Unsampled environments and substrates within the warm-temperate/subtropical U.S. Gulf of Mexico represent a potential source of new taxa and genetic diversity. The roles of substrate, season and latitude in shaping intertidal ascomycete community structure were examined for the U.S. Gulf of Mexico. Best sampling practices were developed and 750 collections of beach detritus, sand and seafoam were made from the Florida Keys north to St. Vincent Island, Florida, from South Padre Island north to Galveston Island, Texas and from West Ship Island, Mississippi. Morphological and molecular techniques (ITS T-RFLP community fingerprinting, ITS gene sequencing) were employed to characterize and compare intertidal ascomycete communities. The role of succession during laboratory incubation was also examined, and this study has increased the number of fungal species known from the Gulf of Mexico by over 50%.

Symposium 4 - Compararive Genomics of Fungi
Day: Wednesday, 18 July
Location:
Time: 1430

: Horizontal gene transfer and public goods games in fungi and fungi-like organisms

Thomas A Richards. Dept. of Zoology. DC1 406C, The Natural History Museum, Cromwell Road, London, SW7 5BD, United Kingdom.

Horizontal gene transfer (HGT) is the transmission of genetic material between organisms, specifically across species boundaries. A growing body of data suggests that fungi and fungal-like protists have gained genes by HGT. This is an exciting result because fungi at first glance represent the most recalcitrant of all organisms to gene transfer, possessing robust cell walls and having lost phagotrophic capacities because they feed exclusively by osmotrophy. Using phylogenetic methods we investigate the role HGT has played in the evolution of fungi and fungal-like protists, including HGT between plants and fungi. This work demonstrates that HGT has actually played a role in shaping osmotrophic phenotypes and furthermore has been important for the evolution of plant parasitic mechanisms in the oomycetes. I then use this data to argue that HGT has also shaped public goods games, an important factor in the evolution of osmotrophic eukaryotes. Together this data suggests HGT in osmotrophic eukaryotes, although relatively rare compared to prokaryotes, seems to be an important factor for shaping the evolution of biological functions in these groups.

Round Table - Toward a roadmap for fungal conservation research in North America: tools, data needs and collaborations
Day: Monday, 16 July
Location:
Time: 1620

: Progress toward a North American Mycoflora

Thomas Bruns. Department of Plant & Microbial Biology, University of California, Berkeley, CA 94720-3102.

Progress on writing a white paper in this year's FESIN workshop - the first step in seeking funding - for a North American Mycoflora will be presented.

Contributed Oral Session 16
Day: Wednesday, 18 July
Location:
Time: 1130

: Soil metatranscriptomics reveals changes in expression of transcripts encoding lignocellulolytic enzymes in the forest floor of a temperate forest under increased nitrogen deposition

Linda TA van Diepen1*, Serita D Frey1, and W Kelley Thomas2. 1Department of Natural Resources and the Environment, University of New Hampshire, Durham, NH 03824, 2Hubbard Center for Genome Studies, University of New Hampshire, Durham, NH 03824.

Fungi are ubiquitous in terrestrial ecosystems and play an important role in biogeochemical cycling because of their function as litter decomposers. It has been demonstrated that increased nitrogen (N) deposition decreases fungal biomass and changes the relative abundance of particular groups and species. In addition increased N can slow litter decomposition and reduce lignolytic enzyme activity. To understand the functioning of fungi under increased N addition in more detail, we performed a soil metatranscriptomic analysis on forest floor samples from a chronic N-addition experiment at Harvard Forest (MA, USA). We extracted total RNA, followed by reverse transcription of poly-A tailed mRNA and Illumina paired-end sequencing of the cDNA. In the forest floor of control and N amended plots, a total of ~460,000 genes were expressed at the time of sampling, of which 172,000 and 95,000 were uniquely expressed in the N addition and control plots, respectively. More than 20,000 genes (~5% of all expressed genes) were either significantly up-regulated or down-regulated in the control versus the N-amended plots (P<0.05). Of these genes, we focused on genes with a medium to high expression value (RPKM >5, Reads Per Kilobase of exon per Million mapped sequence reads) in either the control or N-amended plots. Most of the differences in expression were found within primary metabolic and cellular processes. More specifically, several genes closely related to fungal glycoside hydrolases and laccases, involved in lignocellulose degradation, were found to be down-regulated in the N-amended plots compared to the control plots. Within our study, metatranscriptomics proved to be a useful technique to study the activity of specific genes of interest, while at the same time revealing shifts in the activity of genes that would be overlooked in gene-specific qPCR analysis.

Symposium 7 - Using cellular structure and biochemistry as indicators of fungal phylogeny
Day: Wednesday, 18 July
Location:
Time: 1720

: Biochemical characters support fungal lineages

John D Weete1, Maritza Abril2, and Meredith Blackwell2*. 1Auburn Research & Technology Foundation, Auburn University, Auburn, Alabama 36832 USA, 2Department of Biological Sciences, Louisiana State University, Baton Rouge, Louisiana 70803 USA.

In the mid 20th Century, prominent mycologists began to express the view that fungal biologists had accepted the "strong and growing opinion ... that the fungi may have originated from animal-like forms." Before there were DNA sequences, morphological and biochemical characters (e.g., cell wall components, flagellation, site of meiosis, carbohydrate storage, lysine synthesis, membrane sterols) were important to separate fungi from other organisms and define monophyletic groups. The characters have been useful in establishing the degree of relatedness among organisms, but they have supported lineages that later were validated by DNA analyses. On closer inspection some of the characters are more variable than were appreciated previously. For example the type of sterol, previously thought to be ergosterol for all fungi, actually varies qualitatively within the fungi. Although ergosterol is the major sterol in the vast majority of Dikarya and certain zygosporic fungi (Mucorales, Zoopagales, Dimargaritales), ergosterol is not known in all fungi. In fact, end products of other sterol pathways (cholesterol, 24-methyl cholesterol, 24-ethyl cholesterol, and brassicasterol) occur as major sterols among the lineages. For example members of Pucciniomycota, the earliest diverging Basidiomycota, contain 24-ethyl cholesterol; the few zoosporic fungi tested (Chytridiomycota, Blastocladiomycota) contain a variety of sterols with double bonds only in the C-5 position such as cholesterol and 24-ethyl cholesterol, and Zygosporic groups outside of Mucorales, Zoopagales, and Dimargaritales, also lack ergosterol and contain a variety of C28 sterols. Several studies have used ergosterol as a measure of fungal biomass in the soil, but the method omits arbuscular mycorrhizal fungi (Glomeromycota), which contain 24-ethyl cholesterol as the major membrane sterol. Taxon sampling is poor in certain groups, but biochemical characters hold great promise for looking at lineages as genomics information becomes readily available.

Symposium 1 - Phylogenetic, Ecological, and Functional Diversity of Fungi
Day: Monday, 16 July
Location:
Time: 1350

: Investigating early-diverging fungi from marine and estuarine habitats in North America and Europe

Kathryn T Picard1*, Rowena F Stern2, and François Lutzoni1. 1Duke University, Durham, NC 27708, 2Sir Alister Hardy Foundation for Ocean Science, Plymouth, UK.

Despite increasing efforts to characterize and catalog early-diverging fungi, marine and estuarine habitats remain largely unexplored. Further, traditional culturing methods commonly employed in taxonomic surveys provide only limited insight into the breadth of zoosporic fungal diversity. However, with the application of culture-independent molecular techniques, including environmental cloning and, increasingly, high-throughput sequencing, the presence, distribution, and variety of these previously unculturable microbes is being elucidated. For this study, marine and estuarine habitats in North America and Europe were surveyed for novel fungal phylotypes using next-generation sequencing methods. North American samples originated from a yearlong sampling effort in coastal North Carolina. Monthly sediment and plankton samples were collected from three small estuarine islands located within the Beaufort Inlet in Pamlico Sound. Quarter-annual collections of marine sediments from the shallow waters of Cape Lookout Bight were also made. Using the Ion Torrent platform, the 5' end of the nuclear large subunit (28S) was sequenced from these samples, resulting in ~200 bp reads. European samples originated from open water collections made across the English Channel over the course of three months. Partial nuclear small subunit (18S) rDNA sequences (~200-400 bp) were generated from English Channel samples using 454 pyrosequencing. The conserved 18S and 28S markers were chosen over more variable regions, such as ITS1 and ITS2, based on greater availability of publicly accessible reference sequences. Preliminary analysis of both North American and European sequence data reveals novel fungal phylotypes from across the fungal tree.

Contributed Oral Session 10
Day: Tuesday, 17 July
Location:
Time: 1645

: Xylose-fermenting yeasts from the gut of passalid beetles: geographical distribution

Hector Urbina1*, Janet J Luangsa-ard2, Sung O Suh3, and Meredith Blackwell1. 1Department of Biological Sciences, Louisiana State University, Baton Rouge, Louisiana 70803, USA, 2Phylogenetics Laboratory, BIOTEC, Thailand Science Park, 113 Paholyothin Rd., Khlong 1, Khlong Luang, Pathum Thani 12120, Thailand, 3Mycology and Botany Program, American Type Culture Collection (ATCC), 10801 University Blvd., Manassas, Virginia 20110, USA.

Passalid beetles (Passalidae, Polyphaga, Coleoptera) have a worldwide distribution with approximately 960 species classified in five subfamilies. Our previous studies of gut symbionts of Odontotaenius disjunctus, a common passalid beetle in the eastern USA, showed that several xylose-fermenting (X-F) yeasts are almost always associated with the beetle species. However, similar information on yeast associates of other passalids remains limited. In order to understand more about the associations between these beetles and X-F yeasts, a variety of passalids were collected from Guatemala and Thailand. Over 2500 yeast strains were isolated from about 200 adult beetles belonging to 32 passalid species collected in the two countries. Multi-locus phylogenetic analyses and biochemical characterizations indicated that the X-F yeasts were present in all of the passalid beetles studied. The most notable geographical difference was that the X-F members of the Scheffersomyces clade, especially S. shehatae and S. stipitis, were the most common ascomycete yeasts in the gut of passalids collected in Guatemala. By contrast, Scheffersomyces species, except for Scheffersomyces lignicola, were rare in the gut of passalids from Thailand. The most common Thai yeasts were X-F species in the Spathaspora clade. Based on molecular and other taxonomic characters, several X-F yeasts were determined to be new species of Scheffersomyces and Spathaspora. These results support the conclusion that the X-F yeasts are closely associated with passalid beetles across continents, although further studies with more extensive sampling from additional distant regions will be necessary to understand the nature of the yeast associations with passalid beetles.

This abstract is not yet assigned to a session.

: Insights from comparative genomics in lichen symbiosis

Olaf Mueller1*, Scott Baker3, Guillaume Blanc2, Frank Collart3, Fred Dietrich1, Peter Larsen3, Jon Magnuson3, Francis Martin4, Emmanuelle Morin4, François Lutzoni1, and Daniele Armaleo1. 1Department of Biology, Duke University, Durham, NC 27708 USA, 2Information Génomique et Structurale (IGS), CNRS-UPR2589, IFR-88, Marseille, 3Argonne National Laboratory, 9700 S. Cass Avenue, Argonne, IL 60439, 4IUMR 1136, INRA-Nancy University, Interactions Arbres/Microorganismes, INRA-Nancy, 54280 Champenoux, France.

Mutual recognition and response among plants and fungi are central to either pathogenic or symbiotic associations. Genomes of the lichen-forming ascomycete Cladonia grayi (mycobiont) and its photoautotrophic symbiont, the single-celled green alga Asterochloris sp. (photobiont), were sequenced and analyzed to identify genes specific for the establishment and maintenance of lichen symbiosis. Annotated gene models of C. grayi and Asterochloris were compared to reference genomes representing related Ascomycota taxa (Eurotiomycetes, Leotiomycetes, Sordariomycetes and Dothideomycetes) and Chlorophyta taxa (Chlorophyceae, Trebouxiophyceae and Prasinophyceae), respectively. A mutual best-hit blast approach was performed to identify similarities and differences in gene inventories and to provide a scaffold for detailed phylogenetic analyses. Markov cluster (MCL) studies compiled expanded and contracted gene families in the Cladonia symbionts. Gene families were investigated phylogenetically. Reduced rates of evolution driven by purifying selection are expected for genes essential to the lichen symbiosis, in order to maintain the lichen-symbiosis for a period of more than 400 million years. Candidate genes identified in this manner will be discussed in light of results from transcriptomic analyses derived from RNA isolated from the mycobiont and photobiont grown separately and together in experiments aimed at reconstituting the initial developmental interactions of the lichen symbiosis.

Symposium 4 - Compararive Genomics of Fungi
Day: Wednesday, 18 July
Location:
Time: 1410

: Evolution of plant pathogenesis within Dothideomycetes

Stephen B Goodwin. USDA-ARS, Department of Botany and Plant Pathology, 915 West State Street, Purdue University, West Lafayette, IN 47907-2054.

The Dothideomycetes is the largest class of fungi, occurring on every continent and in very diverse habitats. In addition to many plant pathogens in the orders Capnodiales and Pleosporales, which collectively cause economically important diseases on almost every major crop, Dothideomycetes can endure extremes of heat or cold, high solar radiation, desiccation, live in fresh- or saltwater habitats, persist as lichens or can form mycorrhizal associations with plants. To better understand plant pathogenicity and the genetic basis for this huge ecological diversity, the genomes of more than 20 Dothideomycetes have been sequenced or are in progress, ranging in size from 22 Mb for the extremophile Baudoinia compniacensis to 74 Mb for the banana pathogen Mycosphaerella fijiensis. Genome expansion seems to have occurred primarily by proliferation of retrotransposons, but other mechanisms were noted including the first evidence for transduplication by DNA transposons in fungi. An unusual component of the M. graminicola genome was a "dispensome" of eight chromosomes that can be lost with no obvious effect on fitness, appears to have originated by horizontal transfer more than 10,000 years ago, and is highly variable in field populations. Some other Dothideomycetes genomes have possible dispensable chromosomes that are unrelated to those in M. graminicola. Functional characterization of the genomes revealed that many species of Mycosphaerella have reduced sets of genes for cell wall-degrading enzymes and other proteins, possibly as an adaptation to avoid detection by their hosts to facilitate stealth pathogenicity. Effectors identified in the genome of M. fijiensis by similarity to those in the tomato pathogen Cladosporium fulvum could be recognized by the heterologous resistance gene to elicit a defense response. The extensive genomic resources for the Dothideomycetes provide tools for a comprehensive understanding of the genetics and evolution of adaptations to very diverse environments and modes of nutrition.

Contributed Oral Session 7
Day: Tuesday, 17 July
Location:
Time: 1500

: High throughput analyses of early life history traits of Neurospora crassa

Kolea C Zimmerman1*, Daniel Levitis2, and Anne Pringle1. 1Department of Organismic and Evolutionary Biology, Harvard University, Cambridge MA, 2Max Planck Institute for Demographic Research, Rostock, Germany.

Many organisms experience high mortality during initial growth. This is especially relevant for most fungi because of the large ratio between propagules produced and the number of those propagules that germinate and grow into a mature mycelium. In this study, we developed flow cytometry methods that can be used to determine the fine scale germination dynamics of Neurospora crassa conidia and ascospores. Specifically, we determined germination dynamics of conidia harvested from eight genetically distinct WT strains and we also optimized conditions for the cytometry of ascospores. Conidia and ascospores were germinated in liquid culture and sampled at intervals. Aliquots were fixed and nucleic acids of the germinating spores were fluorescently labeled. Processed samples were measured with a BD Fortessa flow cytometer using a 488 nm laser for all parameters. Spearman rank correlations showed that side-scatter-width and fluorescence-width were the most informative parameters for germination over time. Increased nucleic acid content preceded physical growth and nucleic acid content was the most variable growth trait. This indicates that nucleic acid content and rate of change are informative parameters for further quantitative genetic and heritability analyses. Germination rates and percent germination of conidia differed significantly amongst the wild isolates tested. Finally, we developed a framework to test the effects of parent relatedness on the viability of meiotically produced offspring. We calculated precise genetic distances between environmental isolates of N. crassa using publicly available RNAseq data and devised crosses to represent the range of calculated distances. Our research will facilitate further experimental work on the evolutionary demography of early life history traits in the Fungi.

Symposium 6 - Fungi in a Changing World
Day: Wednesday, 18 July
Location:
Time: 1720

: Tallgrass prairie soil microbial communities are resilient to climate change

Ari Jumpponen. Division of Biology, Kansas State University, Manhattan, KS66506.

Climate models for central United States predict increasing temperatures and greater variability in precipitation. Combined, these shifts in environmental conditions are likely to impact many ecosystem properties and services. The existing Rainfall Manipulation Plots (RaMPs) experiment permits addressing address soil microbial community responses to simultaneous manipulation of temperature and temporal variability in precipitation. The RaMPs experiment is located in the tallgrass prairie at Konza Prairie Biological Station and has been operational since 1998 providing therefore a potential to address soil community responses to environmental manipulations in the long term. To test whether community composition, richness, or diversity respond to environmental change, more than 100,000 bacterial and fungal amplicons across 48 experimental units were 454-sequenced. The acquired data suggest that the soil communities are compositionally resilient to predicted environmental changes. This is the case both for the communities overall as inferred from ordination analyses as well as analyses of variance for each of the most common Operational Taxonomic Units (OTUs). However, while seemingly compositionally resilient, other data sources as well as on-going functional assays strongly indicate functional shifts in these communities.

Contributed Oral Session 17
Day: Wednesday, 18 July
Location:
Time: 1115

: Populus species, genotype, and soil inoculum influences on the assemblage of rhizospheric fungal and bacterial communities

Gregory Bonito1*, Christopher Schadt2, Gerald (Jerry) Tuskan2, Mitchel Doktycz2, and Rytas Vilgalys1. 1Biology Department, Duke University, Durham NC, 2Oak Ridge National Laboratory, Department of Energy, Oak Ridge TN 37831.

Populus deltoides is a common riparian tree species in southeastern North America. Populus forms root associations with both arbuscular and ectomycorrhizal fungi and also with bacterial and fungal endophytes. To address the influence of edaphic and genotypic factors on the structuring of rhizospheric assemblages, we carried out a series of trap-plant experiments using rooted cuttings of various Populus genotypes. Two other tree species (Quercus phellos and Pinus taeda) were included for comparison. Plants were grown in field soils in growth chambers. We used 454 multiplex amplicon pyrosequencing to characterize fungal and bacterial root communities. Specifically, we were interested in the effects of host species, P. deltoides genotype, and soil inoculum on the structuring of rhizospheric communities. Although species richness of endophytic root fungi was higher for Populus than for oak or pine, Populus hosted fewer ectomycorrhizal taxa. Total richness of root associated bacteria taxa (283) was also significantly greater for Populus than for oak (175-f;184-b) or pine (157-f;185-b) and Populus was characterized by a higher relative abundance of Actinobacteriales and Sphingobacteriales, and a lower abundance of Rhizobiales and Burkholderiales. There were minor responses of bacterial and fungal communities to Populus genotype, but the Populus hybrid associated with a wider variety of fungi and a greater frequency and relative abundance of ectomycorrhizal taxa (e.g. Inocybe, Tomentella, Hebeloma). Arbuscular mycorrhizal fungi belonging to the Glomerales and Paraglomales were present in all Populus genotypes but sequences belonging to the Diversisporales were only detected in the hybrid Populus genotype. Soils had a large impact on microbial communities, and abundant species in some soils were absent in others. In conclusion, rhizospheric microbiota of P. deltoides is diverse and unique from other tree species, and appears to be structured both by the microbial inoculum available in soils and, to a lesser extent, by plant genotype.

Symposium 6 - Fungi in a Changing World
Day: Wednesday, 18 July
Location:
Time: 1620

: Mycorrhizae and resource acquisition: dynamics in fluctuating environments

Michael F Allen. Center for Conservation Biology, University of California, Riverside, CA 92521 USA.

Mycorrhizae exist from organic layers deep into soil profiles, even into fracturing bedrock. As soils dry, deeper water either from groundwater or from water bound in cracks and pockets is acquired and utilized or reallocated via hydraulic redistribution. N is acquired from surface soils even under drought conditions. We utilized shifts in natural abundance isotope ratios to better understand spatial and temporal patterns of resource acquisition. Subsequently, utilizing continuous sensor and observation platforms, we monitored shifts in roots, mycorrhizal fungi, and respiration in response to changing temperature and moisture. Acute perturbations consisting of either large storms or drought result in plant mortality, but also subsequently re-order root and fungal symbiosis shifting both the types and spatial structure of resource acquisition. This may be because in seasonal environments, root and fungal activity show complex and often rapid responses to environmental change. We can utilize these shifts to develop a better understanding of belowground response dynamics to more chronic climate change or to acute perturbations such as hurricanes or drought.

Contributed Oral Session 15
Day: Wednesday, 18 July
Location:
Time: 0915

: Description of mycobiomes associated with three species of bats in Illinois

Lynnaun Johnson1*, Shiloh Lueschow1, Jason Stonewall1, Robert McCleery2, Rod McClanahan3, Joseph Kath4, and Andrea Porras-Alfaro1. 1Biological Sciences, Western Illinois University, Macomb, IL 61455, 2Wildlife Ecology and Conservation PO Box 110430, University of Florida, Gainesville, FL 32611-0430, 3USDA Forest Service, Shawnee National Forest, Harrisburg, IL 62946,, 4Illinois Department of Natural Resources, Springfield, IL 62702-1271.

Geomyces destructans is the causative agent of Geomycosis better known as White Nose Syndrome (WNS). This fungus was documented in 2009 as a novel fungal species. Since 2006 it has killed more than a million bats to date with losses in bat populations of 75% or more in the northeastern United States. Though hypotheses suggest that G. destructans originated in Europe, the fungus currently shows no serious threat to European bats. With the high mortality caused by WNS in North America, much focus has been placed on the fungus G. destructans and current literature is lacking detailed analysis about the composition and structure of fungal communities (mycobiomes) in healthy bats. The main objectives of this project are 1) to document fungal flora of hibernating bats in northern IL caves, 2) to determine the presence of other potential Geomyces species and pathogens amongst bats. We sampled 30 bats including Myotis septentrionalis, Myotis sodalis, Perimyotis subflavus from four hibernacula within Illinois (before WNS arrival) and we isolated and sequenced 141 fungi using ITS primers. We found approximately 53 OTUs at 97% similarity. Fungi associated with bats were primarily dominated by Ascomycota (72% of the OTUs). Dominant orders include Eurotiales, Hypocreales, Capnodiales, Pleosporales and Sordariales. Common species isolated from bats included Cladosporium, Geomyces, Fusarium, Mortierella, Penicillium and Trichosporon. Thirteen Geomyces isolates were obtained from different bat species and phylogenetic analysis indicated at least three potential novel Geomyces clades. Temperature tests showed all Geomyces isolates were psychrotolerant, different than G. destructans, a psychrophilic fungus. The widespread distribution of Geomyces among bat mycobiomes can lead to false positives when detecting WNS. The role of these diverse bat mycobiome and their potential pathogenicity and interactions with G. destructans will require further analysis.

Symposium 4 - Compararive Genomics of Fungi
Day: Wednesday, 18 July
Location:
Time: 1350

: Convergent and divergent evolution of nonribosomal peptide synthetases

Kathryn E Bushley* and Joseph W Spatafora. Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97330.

Nonribosomal peptide synthetases (NRPSs) are large multimodular enzymes which produce small bioactive peptides (NRPs) without the aid of ribosomes. While the function of many NRPs remains unknown, it is becoming clear that many help shape the ecologies and lifestyles of their producing fungi. Production of a host-selective toxin, for example, confers the ability to colonize a specific host while production of antibiotics may increase competitive ability in a particular habitat. As such, the ability to produce these compounds is likely under selective pressure. Drawing on examples from across the fungi and focusing on the order Hypocreales, which consists of fungi with diverse ecologies and lifestyles ranging from plant pathogens to insect/animal pathogens, to fungal pathogens, we explore correlations between the evolution of NRPSs biosynthetic pathways, their chemical products, and fungal lifestyles. We examine expansions and contractions of phylogenetic clades of NRPS adenylation domains correlated with lifestyle and address evolutionary and genetic mechanisms that may lead to diversification of chemical products. We also address the extent to which these correlations result from independent expansions versus acquisition by horizontal transfer or non-vertical evolutionary pathways.

Contributed Oral Session 11
Day: Tuesday, 17 July
Location:
Time: 1700

: Multidimensional regulation in transcriptome during sexual development in Neurospora crassa revealed with RNA sequencing

Zheng Wang1, Nina Lehr1*, Francesc Lopez-Giraldez1, Marta Farré2, Frances Trail3, and Jeffrey P Townsend1. 1Department of Ecology and Evolutionary Biology, Yale University, New Haven, CT06520, USA, 22Departament de Biologia Cellular, Fisiologia i Immunologia. Universitat Autònoma deBarcelona. Campus UAB, 08193, Cerdanyola del Vallès, Barcelona, Spain, 3Department of Plant Biology, Department of Plant Pathology, Michigan State University, East Lansing, MI 48824, USA..

Fungi exhibit a huge diversity in morphology and ecology. The genetic basis of morphological development in sexual reproduction of multi-cellular fungi, including the model organism Neurospora crassa, has not been fully investigated with genome-wide approaches. RNA were sampled for eight time points cross perithecial development, including two points right before and after crossing, for which mat A protoperithecia were fertilized with mat a conidia. Transcription profiles for 9717 genes were obtained for all eight time-points using multi-targeting priming and for five time-points using random hexamer priming. The majority of the genome showed continually up- or down- regulated expression patterns during the later perithecial development stages after 72 and 96 h. Functionally unclassified proteins were counted for most up-regulated genes, while genes were enriched with different functions for various down-regulated patterns. We observed a significant increase in transcription for mat a-1 and pheromone precursor ccg-4 during perithecial development. In addition, expression of genetic markers for morphological traits in perithecium, ascus, and ascospore, and for development traits in meiosis often showed multiple peaks during the experiment. Expression patterns for genes involved in Meiotic Silencing and Repeat Induced Point Mutation, and for a gene similar to stc-1, were correlated and correspondent to morphological development. Interesting transcription profiles were also observed for genes involved in heterokaryotic and vegetative incompatibility, in the heterotrimeric G protein signaling systems, and in the MAP kinase signaling pathways. Knockouts of these regulatory genes are being studied for their phenotypes in sexual development of N. crassa.

Symposium 5 - The Fungal DNA Barcode and Beyond
Day: Wednesday, 18 July
Location:
Time: 1330

: Barcodes and a MycoCHIP - a case study on turfgrass pathogen barcoding and diagnosis

Ning Zhang. Dept. of Plant Biology and Pathology, Rutgers University, New Brunswick, NJ 08901.

The internal transcribed spacer (ITS) region of ribosomal RNA genes is one of the most commonly sequenced loci in fungi and has been declared as the official fungal barcode. Usually ITS sequences allow differentiation of fungi at the species level, which makes them ideal for designing species specific probes in molecular detection methods such as DNA diagnostic array. The diagnostic array tool offers a fast, culture-independent alternative for the detection of microbes from field samples. It supports simultaneous detection and differential identification of multiple target species in a single assay. Our current objective is to develop a highly sensitive Turf PathoCHIP for rapid detection of known and emerging turfgrass pathogens, based on ITS sequences of fungi. The development of this product will allow for efficient pathogen identification resulting in more effective disease management, improved plant health and potential reductions in pesticide usage. As the bacterial PhyloChip, this technology can be extended to form a "MycoCHIP", which may cover fungi of all known diversity, in order to detect and identify fungi in other habitats on earth.

Contributed Oral Session 5
Day: Tuesday, 17 July
Location:
Time: 0945

: Taxonomic and phylogenetic evaluation of the holomorphic complex of Cochliobolus, Bipolaris and Curvularia

Dimuthu S Manamgoda1,2*, Lei Cai2, Amy Y Rossman3, Ekachai Chukeatirote1, and Kevin D Hyde1. 1Institute of Excellence in Fungal Research,School of Science,Mae Fah Luang University, Chiang Rai, 57100, Thailand, 2State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100190, P.R. China, 3Systematic Mycology & Microbiology Laboratory, USDA-ARS, Beltsville, MD 20705, USA.

The holomorphic complex of Cochliobolus, Bipolaris and Curvularia are worldwide pathogens of mostly grasses (Poaceae). Some species have caused devastating epidemics of important food crops such as rice, wheat and maize. Therefore, accurate identification of the species is important for disease control and establishment of phytosanitary measures. The taxonomy of Cochliobolus is confusing as frequent nomenclatural changes and refinements of taxonomy have occurred. The two anamorphic genera Bipolaris and Curvularia share many morphological similarities and they are indistinguishable morphologically. Phylogenetically neither Bipolaris nor Curvularia is monophyletic. We investigate the species complex based on the collections from northern Thailand and type cultures and herbaria from BPI and other international collections. Combined gene analysis of rDNA ITS (internal transcribed spacer), GPDH (glyceraldehyde 3-phosphate dehydrogenase), LSU (large sub unit) and TEF (translation elongation factor 1-α) shows that the holomorphic complex divide into two groups. Cochliobolus and Bipolaris clustered in Group I whereas Bipolaris, Cochliobolus, and Curvularia clustered in Group II. All species previously named as Pseudocochliobolus cluster in Group II. The generic type of Cochliobolus and Bipolaris lies in Group I and generic type of Curvularia lies in Group II. Thus, Cochliobolus is selected to represent Group I and Curvularia to represent Group II as they are the older names. Cochliobolus and Curvularia are amended and species used in this study are transferred to the appropriate genera based on phylogeny.

Contributed Oral Session 5
Day: Tuesday, 17 July
Location:
Time: 0915

: Taxonomic and molecular phylogenetic studies of the phytopathogenic genus Diaporthe (Phomopsis)

Dhanushka Udayanga1,2*, Xingzhong Liu2, Amy Y Rossman3, Ekachai Chukeatirote1, and Kevin D Hyde1. 1Institute of Excellence in Fungal Research, School of Science, Mae Fah Luang University, Chiang Rai 57100, Thailand, 2State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences,No 3 1stWest Beichen Road, Chaoyang District, Beijing 100101, P.R. China, 3Systematic Mycology & Microbiology Laboratory, USDA-ARS, Beltsville, MD 20705, USA.

The genus Diaporthe (anamorph Phomopsis) comprises phytopathologically important microfungi with a diverse host association and worldwide distribution. Species recognition of Diaporthe has historically been based on morphology, cultural characteristics and host affiliation. Difficulties in accurate species identification using morphology have led to the application of alternative approaches to differentiate species; these include virulence and pathogenicity, biochemistry, metabolites, physiology, antagonism, molecular phylogenetics and mating. We investigate and evaluate the taxonomy and phylogeny based on specimens collected from northern Thailand, specimens and cultures obtained from BPI and various other collections. The complexes of species that occur on each of the selected hosts and families are also considered. Fresh specimens are identified by means of molecular phylogenetic data based on the ITS rDNA, partial elongation factor 1-α (EF), β-tubulin (TUB), calmodulin (CAL) coupled with morphology. Backbone phylogenetic trees are inferred based on type derived sequences and used to identify the species and define novel taxa and phylogenetic species recognition of Diaporthe is re-evaluated. The older generic name Diaporthe is put forward and several species combinations and new taxa are proposed. The need for epitypification of prevalent phytopathogens and adoption to a single generic name is emphasized.

Contributed Oral Session 11
Day: Tuesday, 17 July
Location:
Time: 1630

: A draft genome of Rhizopogon vesiculosus using the next generation Illumina sequencing platform

Alija B Mujic* and Joseph W Spatafora. Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR, 97331.

The ectomycorrhizal (EM) fungi Rhizopogon vinicolor and R. vesiculosus (Boletales, Basidiomycota) are cryptic sister species of false truffles that form obligate EM relationships with Pseudotsuga menziesii (Douglas Fir). They both form unique tuberculate mycorrhizae and possess a sympatric distribution throughout their range where sampled. Previous and ongoing studies have revealed that while the sporocarp and EM morphology of these fungi may be highly similar, they possess striking life history differences. Rhizopogon vesiculosus produces larger vegetative genets and shows patterns of inbreeding within a patch size of approximately 120 meters. These trends are also reflected at the landscape scale, where R. vesiculosus has been found to display moderate population differentiation. In contrast, R. vinicolor produces smaller genets and shows no patterns of restricted gene flow within sites and only marginal population differentiation at the landscape scale. The frequency and distribution of both EM root tips and sporocarps of these species are relatively equivalent at all sites sampled and surveys of the nuclear count of basidiospores have revealed equal potential for secondary homothallism. These findings afford some explanation of the forces driving divergence between these species, but basic elements of these species' biology such as the mating type system and biogeography remain unknown and limit our ability to test evolutionary hypotheses of sympatric and allopatric speciation. Recent advancements in high throughput sequencing technologies allow for greater applicability of genomic data for population and species-boundaries studies of non-model organisms. In this study we present the partial draft genome of R. vesiculosus as sequenced on the Illumina HiSeq platform. This genome will allow for comparative analysis of gene content and mating type system with other Basidiomycota and ultimately for population/species-level genomic studies between R. vesiculosus and R. vinicolor.

Symposium 1 - Phylogenetic, Ecological, and Functional Diversity of Fungi
Day: Monday, 16 July
Location:
Time: 1450

: A multi-gene pyrosequencing-based approach for elucidating the phylogeny of diverse lichenized microfungi

Brendan P Hodkinson1* and James C Lendemer2. 1International Plant Science Center, The New York Botanical Garden, 2900 Southern Blvd., Bronx, NY 10458-5126, 2Institute of Systematic Botany, The New York Botanical Garden, 2900 Southern Blvd., Bronx, NY 10458-5126.

Lichen-forming fungi represent one of the most readily visible and publicly accessible groups of organisms studied by mycologists. Remarkably, despite their "high profile" nature, many aspects of lichen biology and taxonomy remain poorly understood and critically understudied. These two factors converge noticeably in a highly speciose assemblage of lichens that are referred to as sterile asexually-reproducing crustose lichens, or more simply, "sterile crusts." These taxa do not represent a monophyletic group, but rather are treated together because they have all evolved to reproduce primarily through the dispersal of lichenized diaspores (specialized dispersal units that include the major constituents of the lichen microbiome). Species that employ this mode of reproduction have evolved in nearly all of the diverse lineages that comprise lichen-forming fungi. In an effort to overcome the taxonomic impasse in this group, we have developed a set of high-throughput procedures to test for correlations between molecular sequence data and non-molecular character data to phylogenetically place and phenotypically define a large number of species so that they can be formally described and subsequently integrated into conservation and management plans. This work includes using slower-evolving loci (e.g., mtSSU and nucLSU) to place many of the 'incertae sedis' lichen-forming fungi into a higher-level framework and testing species boundaries using the ITS fungal 'barcode' region.

Contributed Oral Session 15
Day: Wednesday, 18 July
Location:
Time: 0830

: How bad is BAD1? Looking at the relationship between Blastomyces dermatitidis Virulence Factor BAD1 and Human Neutrophils, the "Infantry" of our Initial Immune Response

Angela K Ratekin1*, Bernadette C Taylor1, and Thomas J Volk2. 1Department of Microbiology, University of Wisconsin La Crosse, La Crosse, WI 54601, 2Department of Biology, University of Wisconsin La Crosse, La Crosse, WI 54601.

Blastomycosis, a fungal infection, presents with a variety of disease states. The causative agent, Blastomyces dermatitidis, a thermally dimorphic fungal pathogen, is capable of infecting immunocompetent hosts. The mechanisms of invasion and the evasion of the immune response mounted by the human host are not clearly understood. The broad based budding yeast form is large, making it difficult for the phagocytic cells of the immune system to engulf. Blastomyces ADhesin Factor (BAD1) is a virulence factor associated with the yeast form of B. dermatitidis. Neutrophils are the predominant immune cell involved in phagocytosis and the predominant cell populating the initial defense of the immune system. BAD1 attaches to these phagocytic cells resulting in immunomodulation, a change in chemical messaging. This change may result in an increased activation of neutrophils, resulting in an increase in Neutrophil Extracellur Trap (NET) production. Since the yeast cells are too large to be readily engulfed by the neutrophils, a net with a DNA backbone embedded with antimicrobial peptides and enzymes may aid in the containment and elimination of pathogens. However, with some strains the activation of neutrophils may become overzealous resulting in excess lung tissue damage, presenting as Acute Respiratory Distress Syndrome (ARDS). Looking at isolates with known clinical presentation, BAD1 expression is measured by flow cytometry. The production of NETs is observed by fluorescence microscopy and quantified by measuring elastase, a component of NETs. A percent killed assay is being used to determine the survival of B. dermatitidis post NET production. Our hypothesis is that the level of BAD1 expression correlates with neutrophil activation and NET production, which in turn plays a part in determining the type of clinical manifestation. A high level of BAD1 expression results in tissue damage, ARDS, whereas a lower level results in dissemination of the organism.

Contributed Oral Session 8
Day: Tuesday, 17 July
Location:
Time: 1430

: Ectomycorrhizal fungi associated with seedlings of a monodominant tree in the Neotropics

Gwendolyn C Williams1*, Matthew E Smith2, Terry W Henkel3, and Rytas J Vilgalys1. 1Department of Biology, Duke University, Durham, NC 27708, 2Department of Plant Pathology, University of Florida, Gainesville, FL 32611, 3Department of Biological Sciences, Humboldt State University, Arcata, CA 95521.

The ectomycorrhizal (EM) leguminous canopy tree Dicymbe corymbosa forms monodominant stands in the tropical forests of the central Guiana Shield region. Within these stands, D. corymbosa maintains a large bank of shade-tolerant seedlings that result from mast fruiting events. It has been proposed that common mycorrhizal networks support Dicymbe seedling recruitment and facilitate persistent monodominance, and that an absence of appropriate symbionts may inhibit these seedlings from successfully establishing in nearby mixed forests of AM-associated trees. Little is known, however, about the actual fungi forming mycorrhizae with these seedlings. Using molecular methods we examined EM fungi associated with roots of D. corymbosa seedlings within monodominant stands and along transition zones into AM-dominated mixed forest. Seedlings within monodominant stands hosted significantly higher EM fungal species diversity than those in transition zones, but no effect of distance from parent tree was detected in either location. EM fungi commonly recovered on seedlings included members of the Clavulinaceae, Inocybaceae, Russulaceae, and Thelephoraceae, all of which have been consistently found on adult D. corymbosa roots. The Boletaceae, however, were notably under-represented on seedlings. These preliminary results will be discussed in relation to high-throughput environmental sequencing of soils from the monodominant stands and transition zones.

Contributed Oral Session 3
Day: Monday, 16 July
Location:
Time: 1015

: Comparative transcriptomics identifies new genes important to perithecium development and function in Fusarium

Usha Sikhakolli1, Nicholas Harrison1, Nina Lehr2, Zheng Wang2, Ning Li2, Frances Trail1*, and Jeffrey P Townsend2. 1Department of Plant Biology, Michigan State University, East Lansing, MI, 2Department of Ecology and Evolutionary Biology, Yale Univeristy, New Haven CT.

In recent years, a plethora of genomic sequences have been released for fungal species, accompanied by functional predictions for genes based on protein sequence comparisons. However, identification of genes involved in particular processes has been extremely slow, and new methodologies for identifying genes involved in a particular process have not kept pace with the exponential increase in genome sequence availability. We have performed transcriptional profiling of five species of Neurospora and Fusarium during six stages of perithecium development. We estimated the ancestral transcriptional shifts during the developmental process among the species and identified genes whose transcription had substantially and significantly shifted during the evolutionary process. We examined genes whose expression greatly increased in Fusarium graminearum perithecium development compared to Fusarium verticilloides or compared to Neurospora spp. Functional studies through gene disruption resulted in substantial changes in Fusarium graminearum perithecium morphology in the mutants of many of these genes. These genes were not previously identified as candidates for function in perithecium development, illustrating the utility of this method for identification of genes associated with specific functional processes.

Contributed Oral Session 10
Day: Tuesday, 17 July
Location:
Time: 1630

: A new synnematous hyphomycete parasitic on a Phyllachora species

William RO Soares, Maria DM Santos, Érica SC Souza, and José C Dianese*. DEPARTAMENTO DE FITOPATOLOGIA, UNIVERSIDADE DE BRASÍLIA, 70910900 BRASÍLIA, DF, BRAZIL.

A new Phyllachora species on leaves of an unknown angiosperm was recently found. However, very seldom the stromata of the fungus and its immerse ascomata were free of an absolutely inedited hyphomycete. This new hyperparasite was studied both using light and scanning electron microscopy, and shown to invade the Phyllachora ascomata and the surrounding pseudostromata that resulted from the incorporation of the neighboring plant tissue by the host fungus. The hyphomycete develops into a widespread internal compact hyaline mycelium that later will grow erumpently through the plant epidermis as well as through the melanin-impregnated clypeus of the fungus. Thus, cylindrical compact synnemata showing textura intrincata grow to 285-500 µm long × 57-70 µm diam., completely covered, from base to apex, by a multitude of closely packed verrucose polyphialidic conidiogenous cells (1-2 µm diam.). The conidia [(4)-13-18 × 1-3 µm)] formed are hyaline, mostly 1-septate, elliptical to fusiform. Given the absolutely inedited morphology of the hyphomycete, it becomes clear that it has to be assigned to a new anamorphic genus.

Symposium 6 - Fungi in a Changing World
Day: Wednesday, 18 July
Location:
Time: 1700

: The impact of mycorrhizal fungal traits on ecosystem responses to global change: a modeling approach

Kate H Orwin1*, Miko UF Kirschbaum2, Julie R Deslippe2, and Ian A Dickie3. 1Lancaster Environment Centre, Lancaster University, Lancaster, LA1 4YQ, United Kingdom, 2Landcare Research, Private Bag 11052, Palmerston North 4442, New Zealand, 3Landcare Research, Box 40, Lincoln 7640, New Zealand.

Ecosystems are currently being subjected to multiple global change factors, from nitrogen deposition to increasing CO2 concentrations and changes in temperature. Understanding the consequences of these effects on ecosystem processes such as carbon cycling is of prime importance, as this will determine whether ecosystems are likely to cause positive or negative feedbacks to climate change. Current evidence suggests that carbon cycling can be strongly affected by the traits of the plant community, with most studies focusing on litter quality and plant growth rates. However, recent modelling work has demonstrated that the traits of plant symbionts can also play a significant role in determining ecosystem carbon storage. In particular, the ability of mycorrhizal fungi to directly access organic nutrients can be as important as a major shift in litter quality for carbon storage. The two main mechanisms behind this effect were direct access to organic nutrients causing i) an increase in plant nutrition resulting in enhanced photosynthetic rates and subsequent carbon inputs to the soil and ii) a reduction in microbial access to nutrients, which reduced the rate of soil organic matter decomposition. We have little understanding of the way in which such interactions among mycorrhizal symbionts, their hosts and other soil decomposers may modify the effect of global change factors on ecosystem carbon dynamics. Here, we use a newly developed model (MySCaN: Mycorrhizal Status, Carbon and Nutrient cycling) to examine the impact of global change factors on carbon storage, and how these trends can be affected by mycorrhizal traits. We show that global change effects on carbon storage are not universal, but are modified by the interaction between plants and their mycorrhizal symbionts.

Contributed Oral Session 3
Day: Monday, 16 July
Location:
Time: 0900

: Characterization of MAT genes reveals unexpected patterns of sexual compatibility in Leptographium and Grosmannia

Tuan A Duong1*, Z Wilhelm de Beer2, Brenda D Wingfield1, and Michael J Wingfield1. 1Department of Genetics, Forestry and Agricultural Biotechnology Institute, University of Pretoria, Pretoria 0002, South Africa., 2Department of Microbiology and Plant Pathology, Forestry and Agricultural Biotechnology Institute, University of Pretoria, Pretoria 0002, South Africa..

Species of Leptographium sensu lato (Ophiostomatales, Ascomycetes) are sap-stain fungi vectored by bark beetles (Coleoptera, Scolytinae) and some cause or are associated with root diseases in conifers. Sexual states have been reported for more than 30 species, and following the dual nomenclature system, these have been treated in the teleomorph genus Grosmannia. No sexual state is known for at least 47 additional species and these reside in the anamorph genus Leptographium. The discovery of sexual states for species of Leptographium relies mainly on the presence of fruiting bodies on host tissue at the time of isolation and/or intensive laboratory mating studies, which are often undirected and labor intensive with a low success rate. Very little is known regarding the mating systems in Leptographium and Grosmannia and the aim of this study was to investigate the sexual compatibility of species in these two genera by characterization of the mating type loci. To achieve this objective, we cloned and identified mating type genes in six selected species by screening genomic libraries in combination with long range PCR. Based on these sequences, primers were designed to amplify MAT genes in other Grosmannia and Leptographium species. The primers successfully amplified portions of the mating type genes of more than 30 species and confirmed their thallism, in many species for the first time. Populations of several species were also investigated to obtain evidence of sexual reproduction by comparing the mating type ratio in available isolates. The results suggested that sexual reproduction probably occurs in nature for several of these species, even though their sexual states have never been seen. The mating type genes and primers developed in this study represent important tools to further study the biology and population diversity in the Ophiostomatales.

Contributed Oral Session 14
Day: Wednesday, 18 July
Location:
Time: 0915

: Exploring fungal community structure and function in forest soils: challenges and limitations of current methodologies

Petr Baldrian*, Tomáš Větrovský, Jana Voříšková, Ivana Eichlerová, Jaroslav Šnajdr, Lucia Žifčáková, and Martina Štursová. Laboratory of Environmental Microbiology, Institute of Microbiology of the ASCR, Prague, Czech Republic.

Presently, the structure and function of soil fungal communities receives considerable attention. This attention is fuelled by the recognition of the key role of fungi in the C and N cycling in soils, especially of the forest biomes. In addition, the recent establishment of high-throughput-sequencing methods, labelling with stable isotopes or metaproteomics offers a much higher resolution of the current studies. The interpretation of experimental results is, however, still challenging due to the fact that many methods may potentially contain more or less apparent biasses. This contribution aims to point at the most important limitations of current methodologies to explore fungal abundance, community composition and function in forest soils as studied using biomass quantification techniques (PLFA, ergosterol, qPCR), shotgun or amplicon-based next-generation-sequencing, stable isotope probing and environmental metaproteomics. In the case of microbial biomass surveys, our results show that various methods (rDNA quantification, PLFA or ergosterol assays) yield widely different results of the fungal biomass content or fungal/bacterial biomass ratio and identify the differences in the composition of fungal mycelia / genomes as a source of such errors. For the next-generation-sequencing data, we show that shotgun methods are not consistent with PCR-based methods and that the use of rDNA markers is highly biased due to uneven content of ITS copies per fungal genome as can be demonstrated if single-copy genes are sequenced. The approaches to explore the active fraction of the total fungal community by the analysis of RNA-derived sequencing or the use of Stable Isotope Probing and, most recently, environmental metaproteomics offer an attractive insight into the functioning of fungal communities. However, even the use of these methods must be careful in order to avoid experimental errors.

Contributed Oral Session 4
Day: Tuesday, 17 July
Location:
Time: 0915

: Mycota of the Andean Yungas forests: assessments of fungal biodiversity and habitat partitioning in a threatened ecosystem

Jozsef Geml1*, Eduardo R Nouhra2, Christian Y Wicaksono1, Nicolas Pastor2, Lisandro Fernandez2, and Alejandra G Becerra2. 1Nationaal Herbarium Nederland, Nederlands Centrum voor Biodiversiteit Naturalis, Universiteit Leiden, P.O. Box 9514, 2300RA Leiden, The Netherlands, 2Instituto Multidisciplinario de Biología Vegetal (CONICET), Universidad Nacional de Córdoba, cc 495, cp 5000 Córdoba, República Argentina.

The Yungas, a system of subtropical montane forests on the eastern slopes of the Andes, reach their southern limit in northwestern Argentina. These forests are extremely diverse and, despite covering only 2% of the country's area, they harbour about 50% of Argentina's biodiversity. Unfortunately, the Yungas are among the ecosystems most threatened by anthropogenic pressure and climatic changes. Previous mycological works in the Yungas focused on wood-decay fungi (e.g., polypores) and mycorrhizae in Alnus acuminata cloud forests, while diverse Yungas communities still remain virtually unexplored. We carried out massively parallel pyrosequencing of ITS rDNA from soil samples to provide the first kingdom-wide fungal biodiversity assessment for the Yungas. Samples were taken in the three major forest types along an altitudinal gradient: the piedmont forest (400-700 m asl), the montane forest (700-1500 m asl), and the montane cloud forest (1500-3000 m asl). Using a 97% similarity cut-off value, we delimited 1839 fungal operational taxonomic units (OTUs) in total. The majority belonged to the phylum Ascomycota (52.49%), followed by Basidiomycota (18.85%), Glomeromycota (0.012%), and various zygomycete lineages (0.02%), while 24.91% showed highest similarity to other unidentified environmental sequences and could not be assigned to phylum. The distribution of the total 1839 OTUs among the major altitudinal vegetation types were as follows: 909 OTUs in the piedmont forest, 826 in the montane forest and 929 in the montane cloud forest. Fungal communities were significantly different among all three forest types, with many OTUs showing strong habitat preference for a certain altitudinal zone. Our data offers an unprecedented insight into the fungal biodiversity of the Yungas and into the zonal changes in fungal community structure, with potential applications in conservation strategies to preserve the unique biodiversity of the Andean forests.

Contributed Oral Session 5
Day: Tuesday, 17 July
Location:
Time: 0930

: A phylogenetic study on the Magnaporthaceae (Ascomycota)

Jing Luo* and Ning Zhang. Department of Plant Biology and Pathology, Rutgers University, New Brunswick, New Jersey 08901.

The phylogenetic relationships among taxa in the Magnaporthaceae were investigated based on DNA sequences of multiple genes including ITS, SSU, LSU, RPB1, TEF1 and MCM7. The genera Magnaporthe and Gaeumannomyces were shown to be polyphyletic and their members were divided into four major groups based on the phylogenetic analyses. With the combination of morphological, biological and molecular data, we propose to establish and typify a new genus for the well-supported clade including M. poae, M. rhizophila, and G. incrustans. The current generic concepts of Gaeumannomyces, Magnaporthe and Pyricularia are revised. The constituent species of these genera are listed.

Contributed Oral Session 16
Day: Wednesday, 18 July
Location:
Time: 1030

: Assessing the effects of radiation after the Fukushima Daiichi meltdown on dogwood endophyte communities

Stephen J Miller, Jr.1*, Hayato Masuya2, and Ning Zhang1. 1Dept Plant Biology and Pathology, Rutgers University, New Brunswick, NJ 08901, USA, 2Dept Forest Microbiology, Forestry & Forest Products Research Institute, Matsunosato 1, Tsukuba, Ibaraki 305-8687, JAPAN.

Radionuclide contamination in the environment is one of the predominate concerns to human and environmental health. Radionuclide contamination enters the environment in the form of fallout from nuclear weapons, nuclear waste generated from industries, medical use of radioisotopes, and accidents from nuclear meltdowns. Previously, the role of fungi played in mediating radionuclide movement after the Chernobyl accident was examined. These studies were conducted a few years after the initial accident. Recently, the Fukushima Daiichi nuclear meltdown has released radionuclides in the environment, with radiation levels detectable in North America. In this study, we analyzed the fungal endophytic communities associated with Cornus kousa (Kousa dogwood) leaf samples collected in Japan in June 2010 and June 2011, which was three months after the initial incident. For the June 2010 samples, 320 of 600 leaf segments from 6 samples yielded fungal cultures. For the June 2011 samples, fungi were recovered in 116 of 400 leaf segments from 4 samples located 60-100 km from the Fukushima Daiichi site. In the June 2010 samples, Colletotrichum and Penicillium were the dominant genera. Cosmospora, Phomopsis, and Diaporthe were the dominant genera in the samples recovered after the nuclear meltdown. More melanized fungi were isolated from the 2011 dogwood samples than those collected in 2010. The results indicate that the sampled fungal endophyte communities changed significantly from 2010 to 2011, likely affected by radiation pollution.

Contributed Oral Session 2
Day: Monday, 16 July
Location:
Time: 0915

: Biogeographic and phylogenetic relationships of four easily recognizable morphospecies of Entoloma Section Entoloma (Basidiomycota), inferred from molecular and morphological data

Luis N Morgado1*, Machiel E Noordeloos1, Delia Co-David1, Yves Lamourex2, and József Geml1. 1National Herbarium of the Netherlands, Netherlands Centre for Biodiversity Naturalis, Leiden University, P.O. Box 9514, Einsteinweg 2, 2300 RA Leiden, The Netherlands, 2505, Rue Saint-Alexandre, app. 401, Longueuil (Québec), Canada, J4H3G3.

Species from Entoloma section Entoloma are commonly recorded from both the Northern and Southern Hemispheres and, according to literature, most of them have, at least, Nearctic-Palearctic distribution. However, all records are based on morphological analysis, and studies relating morphology, molecular data and species distribution are lacking. In this study, we selected four morphospecies from Section Entoloma, to answer specific questions considering species concept and geographic distribution: E. sinuatum (E. lividum auct.), E. prunuloides (type-species of section Entoloma), E. nitidum and the European red-listed E. bloxamii, with collections from Europe, North America, Australia and New Zealand. We combined molecular phylogenetics (based on nuclear LSU, ITS and rpb2 and mitochondrial SSU sequences) with morphological analysis to infer interspecific relationships and distribution patterns. Our results indicate that most species appear to have more restricted distribution than previously assumed. None of the collections studied from North Hemisphere and South Hemisphere proved to be conspecific. Entoloma sinuatum and E. nitidum contain phylogeographical groups that are partly recognizable using morphological characters. Entoloma bloxamii is paraphyletic, falling apart into 3 distinct lineages, one represented in Europe, a second restricted to North America, and a third represented in both these continents. The European and North American E. prunuloides appear to be conspecific. Furthermore the accepted taxa appear to belong to two distinct clades, (1) Entoloma clade, with E. sinuatum, E. subsinuatum, E. whiteae, E. flavifolium, and (2) Prunuloides clade, with E. prunuloides, E. bloxamii and E. nitidum.

Contributed Oral Session 9
Day: Tuesday, 17 July
Location:
Time: 1515

: The protist trichomycete genus Enterobryus in the Caribbean

Matias J Cafaro. Department of Biology, University of Puerto Rico, Mayaguez, PR 00681.

Trichomycetes are organisms that live within the digestive tracts of arthropods. Historically, they have been considered one class in the Zygomycota, but molecular studies have determined that the group is not monophyletic. The current phylogenetic position of the orders Amoebidiales and Eccrinales falls within the protist group Ichthyosporea (also known as Mesomycetozoa). Eccrinales is a morphologically diverse order with 17 genera, which inhabit a wide range of hosts: crustaceans, beetles and millipedes in varied habitats (marine, freshwater, and terrestrial). The order is characterized by unbranched, non-septate, multinucleate thalli and sporangiospores that are formed basipetally from the thallus apex. No species have been axenically cultured. Our current effort in the Eccrinales is to understand the diversity and systematics of the largest genus, Enterobryus, typically associated with millipedes. Enterobryus produces two types of sporangiospores, a primary infestation type, which is typically uninucleate, isodiametric and thin-walled, and a secondary infestation type, which is multinucleate, cylindrical and also thin-walled. Species delimitation in this genus is extremely difficult with morphology alone. Intraspecific variation is greater than interspecies variation in some species. In addition, most species descriptions are based on few collections, which do not include details of character variation. In the Caribbean, most islands have endemic millipedes and almost all hosts investigated seem to be infected with Enterobryus. We have been identifying morphological species and including molecular markers to test species hypothesis. Sequence data from multiple clones of 28S rDNA present significant sequence variation within samples indicating high heterogeneity in DNA repeats and hence possible relaxation in the concerted evolution process. New protein markers are needed to generate robust phylogenies.

Contributed Oral Session 7
Day: Tuesday, 17 July
Location:
Time: 1400

: The impacts of 1F:1N on the Ophiostomatales (Sordariomycetes, Ascomycota)

Z Wilhelm de Beer1*, Keith A Seifert2, and Michael J Wingfield1. 1Department of Microbiology and Plant Pathology, FABI, University of Pretoria, Pretoria, South Africa, 2Biodiversity (Mycology and Botany), Agriculture and Agri-Food Canada, Ottawa, Ontario, K1A 0C6, Canada.

The Ophiostomatales includes mainly bark beetle associated fungi, the majority of which are responsible for blue stain of freshly exposed timber, while a few species are tree-killing pathogens. Under the dual nomenclature system, four teleomorph genera were recognized in the Ophiostomatales: Ophiostoma, Ceratocystiopsis, Grosmannia, and Fragosphaeria. Anamorphs in the order were treated in recent years primarily in Sporothrix, Leptographium, Pesotum, Raffaelea and Hyalorhinocladiella, while seven additional anamorph genera were treated as synonyms of these five genera. The adoption of the one fungus one name (1F:1N) principles in the ICN has 'released' the unused genus names for application to newly emerging lineages in the Ophiostomatales, which include species with known teleomorphs. The aims of this study were to consider the impact of the application of the 1F:1N principles on the Ophiostomatales, to redefine genera, to reconsider synonymies and to resurrect generic names where applicable. To achieve this, we re-assessed all published rDNA sequences for the Ophiostomatales, selected the most reliable sequences representing 269 taxa, and conducted phylogenetic analyses using these data. The resulting phylogenies confirmed the monophyly of Ceratocystiopsis and Fragosphaeria, but also showed that Ophiostoma, Grosmannia, and Raffaelea are polyphyletic. The S. schenckii-O. stenoceras- and P. fragrans species complexes, previously treated in Ophiostoma, emerged as distinct genera, namely Sporothrix and Graphilbum. We thus redefined both these genera to include species with teleomorphs previously assigned to Ophiostoma. However, presently available data are insufficient to clarify the generic status of several other lineages currently treated in Ophiostoma s.l., including the status of Leptographium versus Grosmannia, or that of the R. lauricola complex. Because it is presently not possible to propose definitive generic names for some clades, the relevant species are left in their current classification in order to minimize nomenclatural changes, pending the selection of acceptable generic names.

Contributed Oral Session 16
Day: Wednesday, 18 July
Location:
Time: 1100

: The arbuscular advantage: Soil warming stimulates the growth of arbuscular mycorrhizal (AM)-associated tree species and poison ivy over 8 years at Harvard Forest

Jacqueline E Mohan1*, Jerry M Melillo2, James S Clark3, Fahkri A Bazzaz4, Katherine J Bridges1, Charles Cowden1, Paul Frankson1, Richard Shefferson1, Richard Sicher5, Timothy Sipe6, and Lewis Ziska5. 1Odum School of Ecology, University of Georgia Athens, GA 30602, 2The Ecosystems Center, Marine Biological Laboratory Woods Hole, MA 02543, 3Nicholas School of the Environment Duke University Durham, NS 27278, 4Harvard University Department of Organismic and Evolutionary Biology Cambridge, MA 02138, 5USDA, Crop Systems and Global Change Beltsville, MD 20705-2350, 6Franklin and Marshall College Lancaster, PA 17604-3003.

Impacts of warmer soils for nutrient biogeochemical cycling are fairly well known, particularly from soil warming experiments situated on fertile soils of mid-high latitudes; namely, warmer soils cycle nitrogen (N), carbon (C), and in one previous study, phosphorus (P) faster. Potential impacts of warmer soils and increased availabilities of inorganic soil nutrients on plant-mycorrhizal interactions are less clear, as is the scaling up of plant-mycorrhizal relations with warming for ecosystem composition and functioning. Here we present results from the Harvard Forest large-plot (900-m2) soil warming experiment demonstrating that warmer soils with higher availabilities of inorganic N enhance the growth of arbuscular mycorrhizal (AM)-associated woody species with important implications for future carbon sequestration by forests. In the pre-treatment growing seasons of 2001 and 2002, juvenile trees had similar growth rates in the to-be-Heated and the Control plots. Following warming, commenced May 2003, juvenile trees associating with AM fungi displayed significant and consistent growth increases in the Heated plot. This enhanced growth was sustained over time (2003-2010) and by the following taxa: sugar maple (Acer saccharum), red maple (A. rubrum), black cherry (Prunus serotina), white ash (Fraxinus americana), and poison ivy (Toxicodendron radicans). Co-dominant oaks, pines, and birches did not grow faster with warming. Further, mycorrhizal root colonization was significantly less in the maple species growing under warming while foliar %N and %P values were maintained, suggesting faster growth of AM-species may be due to decreased C costs of mycorrhizal associates. These results suggest a future shift in eastern temperate composition towards less productive tree species with important implications for future carbon dynamics. These finding may partially explain the increasing abundance of red maple and woody vines observed over the last several decades in eastern forests and globally, respectively.

Contributed Oral Session 14
Day: Wednesday, 18 July
Location:
Time: 0945

: Fungal and bacterial diversity in connection with soil decomposition processes resolved using 13C rRNA-SIP

Julia Moll1,2*, Dörte Dibbern3, Susanne Kramer4, François Buscot1,2, Dirk Krüger1, Tillmann Lueders3, Sven Marhan4, and Ellen Kandeler4. 1UFZ - Helmholtz Centre for Environmental Research, Department of Soil Ecology, Th.-Lieser-Str. 4, D-06120 Halle/Saale, Germany, 2University of Leipzig, Institute of Biology I, Johannisallee 21-23, D-04103 Leipzig, Germany, 3Helmholtz Zentrum München, German Research Center for Environmental Health, Institute of Groundwater Ecology, Ingolstädter Landstr. 1, D-85764 Neuherberg, Germany, 4University of Hohenheim, Institute of Soil Science and Land Evaluation, Soil Biology Group, D-70593 Stuttgart, Germany.

Soil fungi and bacteria play an important role in global nutrient cycles because of their decomposition of organic materials. The use of rRNA-based stable isotope probing (SIP) allows us to follow the carbon flux into soil microbial communities. Using this novel approach, we aimed at quantifying the carbon flow from plant-derived substrates into the bacterial and fungal channel at different stages of decomposition and to link these microorganisms to specific degradation processes. Highly 13C-labeled labile (glucose) and recalcitrant (cellulose, maize roots, maize leaves) substrates were mixed into soil microcosms and incubated for 32 days. Following cesium chloride density gradient ultracentrifugation of RNA extracts (separating heavy labeled from light unlabeled RNA) from controls and 13C-microcosms sampled at 2, 8, 16 and 32 days post-incubation, rRNA was converted to cDNA and amplified. T-RFLP on the SSU and 16S amplicons was applied to detect the fungi and bacteria utilizing these substrates. It was possible to identify carbon assimilating microbes for all substrates and time points representing substrate degradation stages. For both fungi and bacteria, the community (of operational taxonomic units, OTUs) that consumed carbon from labile substrate clearly differed from that consuming it from recalcitrant material. The bacterial communities that appeared to be actively assimilating carbon in an early stage of decomposition differed from that active in later stages. The fungal utilizers, however, rather remained the same over time. To validate our results and to better identify active taxa, clone library or pyrosequencing approaches are currently added onto the analysis of our experiment. By using SIP, this study will thus gain first insights into the relationship between carbon quality and availability and it will link soil microbial taxonomic with functional diversity.

Contributed Oral Session 7
Day: Tuesday, 17 July
Location:
Time: 1445

: Sterile asexually reproducing crustose lichens: improving classification, conservation, and communication in the 21st century

James C Lendemer1* and Brendan P Hodkinson2. 1Institute of Systematic Botany, The New York Botanical Garden, Bronx, NY 10458-5126, U.S.A., 2International Plant Science Center, The New York Botanical Garden, Bronx, NY 10458-5126, U.S.A...

Although many lichens reproduce through the dispersal of sexual diaspores, asexual reproduction is not uncommon and occurs in nearly all of the diverse lineages that comprise lichen-forming fungi. This type of reproduction can occur through either purely fungal diaspores or lichenized diaspores. The latter comprise a morphologically diverse array of structures that have evolved to facilitate the co-dispersal of the components of the lichen microbiome. The majority of crustose lichens that reproduce exclusively via lichenized diaspores are poorly understood, under collected, or undescribed and they represent a large component of lichen biodiversity that is currently overlooked and underestimated. We present a conceptual summary of the results of five years of research on this group in the context of a revision of the sterile crustose genus Lepraria s.l. In addition to extensive morphological studies of thallus ultrastructure, and integrated studies of molecular and non-molecular datasets to resolve species boundaries, we show that Lepraria s.l. is polyphyletic at the ordinal level and contains disparate elements belonging to four families in three orders, one of which was previously unrecognized. Drawing upon this body of work we present a standard protocol to improve our understanding of sterile crustose lichens, communicate this understanding to a diverse array of interdisciplinary researchers, and translate this understanding into meaningful applications to promote real-world conservation.

Symposium 2 - The science that underpins fungal conservation
Day: Monday, 16 July
Location:
Time: 1450

: Current fungal data from the Great Smoky Mountains ATBI are inadequate to determine conservation status: what is needed?

Karen W Hughes* and Ronald H Petersen. Ecology and Evolutionary Biology, University of Tennessee, Knoxville, TN 37996.

Rapid urbanization and deforestation in the Southeastern US has heavily impacted native plants, and many remain only in small reserves. While there is no similar assessment for fungi, it is likely that fungal species follow similar patterns of fragmentation and localized extirpation. Determining endemism and rarity, however, is significantly hampered by a lack of focused species-level research and data concerning fungal distribution patterns. The Great Smoky Mountains National Park (GSMNP) is a protected reserve that exhibits high biological diversity for fungi, salamanders and plants. The GSMNP and adjacent regions of the southern Appalachian Mountains provide an opportunity to identify remnants of native southeastern mycota. We report studies at the species level which identify putative endemics for further study, examine distributions in so far as is possible and note high levels of cryptic speciation in fungi which may influence estimates of richness and endemism.

Contributed Oral Session 11
Day: Tuesday, 17 July
Location:
Time: 1645

: Comparative genomics reveals complex patterns of secondary metabolite evolution in the genus Elaphocordyceps

Alisha C Owensby*, Kathryn E Bushley, and Joseph W Spatafora. Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR.

Hypocreales is an order characterized by a dynamic evolutionary history of interkingdom host jumping, with members that parasitize animals, plants, and other fungi. The monophyly of taxa attacking members of the same kingdom has not been supported by molecular phylogenetics, however. For example, Trichoderma spp. and Elaphocordyceps spp. are both mycoparasitic, but are members of two different families within the Hypocreales, the Hypocreaceae and Ophiocordycipitaceae, respectively. In fact, both species are more closely related to insect pathogens, than they are to each other. Three species of Trichoderma have sequenced genomes, and more recently the genomes of several insect pathogens in the Hypocreales have been published (e.g. Cordyceps militaris, Metarhizium anisopliae, M. acridum) and others are in progress (e.g. Tolypocladium inflatum). Elaphocordyceps is a genus within this clade that primarily parasitizes ectomycorrhizal truffles in the genus Elaphomyces, representing a yet unsampled ecology within Hypocreales. To compare gene space of a truffle pathogen with closely related insect pathogens and distantly related mycoparasites, we sequenced the genome of Elaphocordyceps ophioglossoides. Our draft assembly of the E. ophioglossoides genome has a size of approximately 32 MB and 10,779 predicted genes. Here, we present a survey of E. ophioglossoides genes involved in secondary metabolism. Notably, E. ophioglossoides is missing the cyclosporin producing non ribosomal peptide synthetase (NRPS) found in its congener, T. inflatum, but possesses the destruxin NRPS, recently identified in Metarhizium anisopliae. The three largest genes in E. ophioglossoides are related to peptaibol NRPS genes, which are only described from Trichoderma spp., and are indicated as being important to mycoparasitism. Our work represents the first genome of an Elaphocordyceps species parasitic on other fungi and will provide insights into mechanisms underlying mycoparasitism and interkingdom host jumping.

Contributed Oral Session 17
Day: Wednesday, 18 July
Location:
Time: 1100

: Endophyte and pathogen communities of symptomatic and asymptomatic seedlings of tree species from a temperate forest

Maria S Benitez1*, Michelle H Hersh2, Brantlee S Richter3, Rytas Vilgalys4, and James S Clark1. 1Nicholas School of the Environment, Duke University, Durham, NC, 2Program in Biology, Bard College and Cary Institute of Ecosystem Studies, Annandale-on-Hudson, NY, 3Plant Pathology, University of Florida, Gainesville, FL, 4Department of Biology, Duke University, Durham, NC.

Fungal pathogens can impact plant community composition through negative density dependence regulation. Pathogen identification is required to quantify their effects on plant host species and diversity. To identify potential seedling pathogens we compared fungal communities associated with symptomatic and asymptomatic seedlings from seven plant species sampled at the Duke Forest (NC) on multiple years. Surface sterilized plant tissue was used for fungal pure culture isolation and total DNA extraction for community analysis. Taxonomic identification of pure cultures was performed based on sequence information of the internal transcribed spacer region (ITS). A collection of over 1300 pure cultures, classified into 316 fungal taxa (97% similarity at the ITS region), was generated from 320 seedlings. The most abundant fungal groups present in symptomatic seedlings differed from those most commonly isolated from asymptomatic seedlings and often represent known plant pathogenic groups. In addition, the most abundant pathogen taxa were isolated from five of the seven host species studied, and were isolated at lower frequency from asymptomatic seedlings. The most abundant isolates from asymptomatic seedlings most closely matched samples from GenBank with no clear taxonomic identification. For individual seedlings studied, the fungal community appears to be simple at this stage, dominated by one or two taxa. At the isolate level, no host-specificity was observed; however, preliminary results from culture-independent analyses reveal community structure separation according to host species. The implications of host specificity of fungal pathogens in forest seedlings are discussed.

Contributed Oral Session 12
Day: Tuesday, 17 July
Location:
Time: 1600

: Research Coordination Network for US culture collections of microorganisms associated with plants

Kevin Mc Cluskey1* and Jessie Glaeser2. 1University of Missouri-KC, 5100 Rockhill Road, Kansas City, MO 64110, 2Center for Forest Mycology Research, USDA-Forest Service, NRS,Forest Products Laboratory, One Gifford Pinchot Drive, Madison, WI 53726.

A network bringing together scientists working with laboratory based collections of microbes is being established. The network will hold numerous workshops teaching best practices for managing, preserving, and distributing bacteria, fungi, and other microscopic organisms in the context of formal culture collections. Biosecurity and regulatory issues will also be emphasized at workshops. Additional goals include re-establishing a professional society of culture collection researchers in the US, developing internet based collection management tools, and fostering communication between US collections, foreign collections, and international collection networks. Collections of living microbes assure that current and past research and innovation are available to future generations of scientists. Biological materials which are made available via well managed collections represent the foundation of the modern biotechnology industry. Materials in culture collections impact fields as diverse as human health, agricultural productivity, biotechnology, and biodiversity research. While there are several professional collections in the US, most collections in the US are small and do not have long term strategies for survival. The network supported by this grant will ensure that smaller collections benefit from expertise available at larger living microbe collections. NSF has recommended our RCN proposal for funding, but funding has not yet been approved. All interested parties are invited to participate.

Contributed Oral Session 14
Day: Wednesday, 18 July
Location:
Time: 0900

: Nutrient enrichment increased species richness of leaf litter fungal communities in a diverse lowland tropical forest

Jennifer F Kerekes1*, Michael Kaspari2, Bradley S Stevenson3, and Thomas D Bruns1. 1111 Koshland Hall, Dept. of Plant and Microbial Biology, UC Berkeley, Berkeley CA 94720, USA, 2Graduate Program in EEB, Dept. of Zoology, University of Oklahoma, Norman, OK 73019, USA, 3Graduate Program in EEB, Dept. of Botany and Microbiology, University of Oklahoma, Norman, OK 73019, USA.

We explored leaf litter fungal diversity in a diverse lowland tropical forest in which a replicated factorial N, P, K, micronutrient fertilization experiment of 40 x 40 m plots had been on going for nine years. Fertilized plots were compared with control plots that did not receive fertilization to evaluate possible nutrient effects on leaf litter fungal communities. We extracted DNA from leaf litter samples collected on the Gigante peninsula, Barro Colorado Nature Monument (BCNM), Republic of Panama and used fungal-specific amplification and a 454 pyrosequencing approach to sequence two loci, the nuclear ribosomal internal transcribed spacer (ITS) region and the nuclear ribosomal large subunit (LSU) D1 region. Overall, both the ITS and LSU gene regions had similar results. The use of two separate gene regions for the analysis provided a complimentary look at the data. Ascomycota (including approximately 29 orders, 44 families, and 88 genera) are the dominant phylum among the leaf litter fungi, followed by Basidiomycota (including approximately 31 orders, 38 families and 52 genera). The long-term addition of nutrients increased species richness relative to the control plots, and had an effect on the taxonomic composition of the leaf litter fungal communities at lower taxonomic levels (i.e. family, genus and species), but not at higher taxonomic levels (i.e. phylum, class and order). Nitrogen has the largest effect, followed by phosphorus and potassium.

This abstract is not yet assigned to a session.

: Three unfinished mycological tales

David S Hibbett. Biology Department, Clark University, 950 Main Street, Worcester, MA 01610.

My mycological interests concern three broad topics: evolution of fruiting body forms, evolution of nutritional modes, and classification. I am particularly focused on mushroom-forming Fungi (Agaricomycotina). Central to my work in all these areas is a phylogenetic perspective gained through analyses of molecular characters. In this presentation I will attempt to summarize some of what I think I have learned, and the questions and challenges that remain.

Contributed Oral Session 3
Day: Monday, 16 July
Location:
Time: 0845

: Partial reconstruction of the ergot alkaloid pathway in Aspergillus nidulans

Katy L Ryan* and Daniel G Panaccione. West Virginia University, Division of Plant and Soil Sciences, Morgantown, WV 26505.

Ergot alkaloids are pharmaceutically and agriculturally important secondary metabolites produced by several species of fungi. A better understanding of the biosynthetic pathway of ergot alkaloids may allow for modification of these compounds to produce effective pharmaceuticals or control their accumulation in agriculture. Ergot alkaloid pathways vary among fungi, but the pathway intermediate chanoclavine appears to be evolutionarily conserved among ergot alkaloid producers. Chanoclavine synthesis occurs in at least five steps, and knockout and heterologous expression studies have demonstrated that four genes, dmaW, easF, easE, and easC, are required for pathway steps prior to chanoclavine. However, it is currently unknown whether these genes are sufficient to synthesize chanoclavine. DmaW prenylates tryptophan to produce dimethylallyltryptophan (DMAT), and easF encodes a methyl transferase that produces the second intermediate, N-Me-DMAT. The roles of EasE and EasC have not been established. Four genes, dmaW, easF, easE, and easC were amplified from the human pathogen Aspergillus fumigatus and transformed into Aspergillus nidulans, which does not contain any of the ergot alkaloid synthesis genes and produces no ergot alkaloids. We determined by HPLC and LC/MS that the A. nidulans strains containing the four A. fumigatus genes produced chanoclavine and earlier pathway intermediates. The specific roles of EasE and EasC are being investigated through the generation of transformants containing DmaW, EasF, and EasC, or DmaW, EasF, and EasE, compared to appropriate controls. HPLC and LC/MS data demonstrated that EasC catalyzes the formation of a new intermediate; the role of EasE is still being investigated. We conclude that the four genes listed above are sufficient for the synthesis of chanoclavine in A. nidulans and that our approach of expressing ergot pathway genes in A. nidulans will provide a better understanding of the early steps in ergot alkaloid synthesis.

Contributed Oral Session 4
Day: Tuesday, 17 July
Location:
Time: 0945

: Assessing fungivory in rodents, marsupials, and kiwi from stable isotope evidence

Erik A Hobbie. Earth Systems Research Center, University of New Hampshire, Durham, New Hampshire, USA.

Fungivory is both a common dispersal mechanism for many fungi and an important source of nutrients and energy for many small animals, particularly rodents and marsupials. Studies of fungivory generally rely on scat or gut content analyses. An alternate technique, stable isotope measurements on animal protein, relies on sporocarps of ectomycorrhizal fungi having distinctive carbon and nitrogen isotope ratios (13C/12C and 15N/14N, expressed as δ13C and δ15N) compared to other ecosystem components. This approach allows retrospective studies by using archived museum specimens; here, I present three case studies of this approach. In studies of western Oregon rodents, mean isotopic values for the five sampled taxa separate into three main groups, with creeping vole and red tree vole quite similar (low in δ13C and δ15N, herbivorous), red-backed vole and flying squirrel similar (high in δ13C and δ15N, indicating high levels of fungivory), and Douglas squirrel different from the other two groupings (high in δ13C, low in δ15N, probably primarily a seed-eater). In ten genera of Australian marsupials, potoroos and bettongs appeared highly fungivorous, as expected from prior dietary studies. Kiwi in New Zealand may perform the same dispersal function for truffles as small mammals do elsewhere. Kiwis were isotopically distinct from frugivorous and insectivorous birds, but truffle consumption probably cannot be distringuished isotopically from consumption of soil invertebrates. Isotopic analysis appears to be a potentially useful tool to determine levels of fungivory in historic and current animal populations.

Contributed Oral Session 10
Day: Tuesday, 17 July
Location:
Time: 1745

: Pathogenesis and host response during infection of seeds by Aspergillus flavus and Fusarium verticillioides

Xiaomei Shu. Department of Plant Pathology, Box 7567, 267 Partners III, North Carolina State University, Raleigh, NC 27695-7567.

Both Aspergillus flavus, a saprophyte, and Fusarium verticillioides, an endophyte, can colonize developing maize seeds and contaminate them with mycotoxins. Susceptibility is conditioned by plant stress, but little is known about the infection processes by the two fungi, and whether their different trophic lifestyles affect tissue colonization and host response. To unravel the biology and pathogenesis of these two fungi, we followed fungal infection and host defense response of developing maize seed for three days after introducing the conidial suspension through the seed pericarp. Histological examination of thin sections of maize seeds showed that the fungi were restricted to the site of inoculation 48 hours post inoculation. Within the next 24 hours mycelium was detected in the aleurone, the endosperm and the embryo-endosperm interface. Mycelium of A. flavus was observed on the surface of either enlarged or shrunken cells of the aleurone, but never inside intact cells. In contrast, F. verticillioides was sometimes observed inside intact aleurone cells. RNA in situ hybridization showed the transcriptional induction of two maize defense genes, PRms (Pathogenesis related protein, maize seeds) and UGT (UDP-glucosyltransferases), in specific seed tissues in advance of visible fungal colonization, suggesting that the fungal virulent factors or plant defense signaling transduction molecules may be involved. Induction of gene transcription often appeared first in the tip of scutellum, which is the cotyledon of the seed. This observation suggests that the scutellum may be very important in plant-microbe communication. Even though these two fungi are characterized as having different trophic lifestyles, they follow a similar pattern of colonization, and induce two known defense genes in maize seeds. Perhaps structural or chemical characteristics of seeds favor a common path of colonization, or the two fungi share a common tool box for the infection of maize seeds.

Contributed Oral Session 8
Day: Tuesday, 17 July
Location:
Time: 1445

: Synopsis of Boletaceae from the central Guiana Shield

Dillon R Husbands1* and Terry W Henkel2. 1Department of Agriculture, Faculty of Agriculture and Forestry, University of Guyana, Georgetown, Guyana, 2Department of Biological Sciences, Humboldt State University, Arcata, CA 95521.

Mycological exploration in the remote tropical forests of Guyana, in the central Guiana Shield phytogeographic region of northeastern South America, has uncovered upwards of 30 species of the pore mushroom family Boletaceae sensu lato. Boletes have heretofore been poorly documented from the South American tropics. These species are distributed across eight sensu lato genera including Austroboletus, Boletellus, Chalciporus, Fistulinella, Phylloporus, Pulveroboletus, Tylopilus, and Xerocomus. Eighteen of these species, most of which were new to science, have been formally described. Molecular analysis of roots has confirmed that at least sixteen of these bolete species are ECM associates of the Fabaceae hosts Dicymbe spp., Aldina insignis, the endemic dipterocarp Pakaraimaea dipterocarpacea, or Coccoloba spp. (Polygonaceae). We will discuss current work on new species of Xerocomus from this bolete-rich region.

Contributed Oral Session 13
Day: Wednesday, 18 July
Location:
Time: 0930

: Alaska - Europe: Biogeography of arctic populations of Microbotryum

Chris Eberlein, Angela M Schäfer, and Dominik Begerow*. Ruhr-University Bochum, Geobotanik, ND03/174, 44780 Bochum, Germany.

Climatic changes over millions of years have shaped the vegetation of our earth. Especially plants of extreme habitats were heavily influenced and many species are extinct due to various ice ages in Europe. While there have been several studies on the population structure of higher plants of artic and alpine distribution, very little is known about the associated, host specific parasites. The aim of this study is the establishment of microsatellites for Microbotryum silenes-acaulis based on low coverage genome sequencing and the analysis of populations from the Alps, Scandinavia and Alaska. Although the coverage is low, genome sequence data revealed hundreds of microsatellites of various size. While the number of bi- and trinucleotid repeats seems somehow limited, there are especially many longer motives of up to nine basepairs repeated many times. To differentiate neighbouring populations as well, we used highly variable satellites to analyse the populations structure of alpine and arctic populations. The link of M. silenes-acaulis collected one year ago during the MSA meeting in Fairbanks to European alpine or arctic populations will be discussed.

Symposium 2 - The science that underpins fungal conservation
Day: Monday, 16 July
Location:
Time: 1430

: Population genetics of Lobariaceae of conservation importance

Christoph Scheidegger. Swiss Federal Institute for Forest, Snow and Landscape Research, WSL, Zürcherstr. 111, CH-8903 Birmensdorf, Switzerland.

Population dynamics of epiphytic lichens depend on the dynamics of their phorophytes and forest stands where they occur. Spatio-temporal connectivity of habitats largely determines the spread of epiphytic lichens among trees within and among stands. Especially lichen species with relatively large, often symbiotic propagules which are often dispersal limited follow a patch-tracking metapopulation dynamics and therefore a loss of spatio-temporal connectivity of habitats is a major cause for population decline of members of this guild. Forest management and catastrophic forest stand disturbance affect epiphytic lichens, especially species belonging to the family Lobariaceae, which often reproduce with relatively large symbiotic propagules, depend on the availability of old trees and are associated with rare green-algal and/or cyanobacterial photobionts. Population bottlenecks, founder events and range expansions leave their genetic footprints in lichen population. We have studied effects of forest management practices on population genetics of Lobaria pulmonaria in a traditional mountain forest landscape. A severe stand level disturbance event 140 years ago led to a drastically reduced genetic diversity in the L. pulmonaria population that has re-established compared to populations where a continuous forest is managed by single-stem harvest. A comparison between L. pulmonaria populations growing in primeval forest reserves and managed forests revealed a reduced genetic diversity in managed forests. Limited habitat availability and a lowered functional connectivity of old trees in managed stands are major causes for declining populations and reduced genetic diversity. Currently a high percentage of Lobaria populations in Swiss forests are restricted to less than five trees. These populations face an increased risk of local extinction and can suffer from an unbiased distribution of MAT-idiomorphs. Lack of one of the two idiomorphs will make sexual reproduction in this heterothallic species impossible unless the complementary idiomorph can be added by re-introduction, as will be shown in an example.

Contributed Oral Session 14
Day: Wednesday, 18 July
Location:
Time: 0930

: Fungal diversity associated with differing monoculture forest types measured by next-generation metabarcoding.

Joshua R Herr. 321 Forest Resources Building, School of Ecosystem Sciences, The Pennsylvania State University, University Park, PA 16801.

It has long been understood that fungi contribute to many key ecosystem processes. This is particularly important in forest soils, where saprotrophic fungi are the main drivers of plant organic matter decomposition and ectomycorrhizal fungi facilitate nutrient uptake for the host plants, thereby affecting plant growth and fitness. Additionally, the presence of soil fungi shape seedling establishment and evidence suggests that ectomycorrhizal fungi may contribute to the distribution of photosynthate from one tree to another, directly regulating the survival of nurse seedlings. Despite their important ecological roles, there is a paucity of information regarding taxonomic diversity and abundance associated with plant host. Largely this is due to the fact that many fungi are unculturable, lack known sexual structures, and are recorded only by environmental sequencing. In order to understand the diversity of fungi associated with differing forest types, we have utilized next-generation sequencing techniques (454/Roche and Illumina/Solexa) to probe soil fungal diversity. Seven mono-culture forest types (Fagus, Quercus, Pinus, Pseudotsuga, Picea, an "old-growth" native forest dominated by Fagus, and an early successional Populus bio-energy plantation) were sampled. Next-generation sequencing showed extreme diversity of fungal taxa. Only 24% of taxonomic sequence reads were able to be identified with existing fungal databases. This finding agrees with recent studies employing new sequencing techniques to probe fungal diversity. Our sequencing shows and average of more than 2000 unique fungal taxa in as little as four grams of forest soil. This presentation will discuss diversity across forest type surveyed and across seasonal sampling.

Contributed Oral Session 13
Day: Wednesday, 18 July
Location:
Time: 0915

: Population dynamics of aerial and terrestrial populations of Phytophthora ramorum in a California watershed under different climatic conditions

Catherine A Eyre, Melina Kozanitas, and Matteo Garbelotto*. Forest Mycology and Pathology Laboratory, Department of Environmental Science, Policy and Management, University of California, Berkeley, CA 94720, USA.

In the attempt to better understand the epidemiology of Sudden Oak Death in California, we present the first combined genetic analysis of soil and leaf genotypes performed for a plant pathogen. Isolates of Phytophthora ramorum were collected from leaves and soil in six plots during a drought and an average-rainfall year, and analyzed using polymorphic SSRs. Leaf populations were re-sampled in different seasons during both years to provide information on how seasonal weather patterns may affect population dynamics. Confirmed SOD symptoms on leaves of the transmissive host Bay Laurel increased from 15 to 39% between dry and wet conditions. Symptoms caused by other foliar pathogens were highest (69%) in dry conditions, suggesting that, while P. ramorum and other pathogens occupy the same niche, they are favored by different climatic conditions. Some foliar genotypes of P. ramorum were more abundant in wet than in dry conditions and persistent through time. Conversely, persistence of soil genotypes was limited. Soil and foliar populations were genetically distinct, (FST>0.15), but not segregated into different portions of a minimum spanning network, suggesting intermixing of the two. We surmise that the genetic structure between substrates results from genotype-specific differences in substrate adaptation rather than lack of gene flow; this hypothesis is supported by different genotype rank order abundances between soil and leaf populations. In climatic conditions unfavorable to the pathogen, genetic diversity increases both within and between sites, while in favorable conditions diversity decreases due to greater migration levels of some genotypes. Foliar genotypes can spread farther in wet years, and soil appears to be re-inoculated on a yearly basis. Cumulatively, our results indicate that leaves act as a relatively persistent source of inoculum, while soil is a sink that is genetically very diverse, but potentially inconsequential for the natural spread of the disease.

Contributed Oral Session 10
Day: Tuesday, 17 July
Location:
Time: 1730

: Temporal fluctuations and the role of disturbance in disease progression of the Sudden Oak Death epidemic

Melina Kozanitas*, Todd W Osmundson, and Matteo Garbelotto. Forest Mycology and Pathology Laboratory, Department of Environmental Science, Policy and Management, University of California, Berkeley, CA 94720, USA.

With its high host mortality and ability to cause landscape-scale alterations in forest cover and composition, Sudden Oak Death (etiological agent ­Phytophthora ramorum, Stramenopila, Oomycota) mirrors past forest disease epidemics such as Chestnut Blight and Dutch Elm Disease. In contrast with these past epidemics, however, the appearance of Sudden Oak Death converges with a time of significant advancement in the development of molecular genetic tools that allow the movement of individuals (or individual genotypes) to be tracked and the role of evolutionary processes in disease progression to be assessed. Such methods have been instrumental in reconstructing the likely origins and geographic pathways of spread of the pathogen; however, little is yet known about the local-scale processes contributing to disease maintenance and progression. Since 2008, we have surveyed a network of sampling plots in the San Francisco Bay Area multiple times per year in order to examine seasonal patterns in the incidence and population genetic structure of infection foci. A period of severe drought in 2007-2009 followed by a period of normal to above-normal precipitation in 2010-2011 provide a significant opportunity to examine the effects of abiotic disturbance on disease progression. Here, we present recent and ongoing research focused on assessing seasonal patterns in genotypic diversity of viable infections that can act as reservoirs of new infectious propagules, assessing the effect of drought as a potential agent of selection on pathogen genotypes, examining the relationship between infections on dead-end (Oak) and amplifying (California Bay Laurel) hosts, and assessing the possible role of competitive interactions with sympatric Phytophthora species (P. pseurosyringae, P. nemorosa) using culture-based surveys, culture-independent (qPCR) assays, and analyses of population genetic structure (based on variable microsatellite loci) to infer the underlying processes of pathogen demographic expansion, contraction and spread.

Contributed Oral Session 3
Day: Monday, 16 July
Location:
Time: 0915

: Mixed mating and biparental inbreeding in the forest pathogen, Neonectria ditissima

Robert E Marra. Department of Plant Pathology and Ecology, The Connecticut Agricultural Experiment Station, 123 Huntington Street, New Haven, CT 06511.

Neonectria ditissima is an important endemic forest pathogen of eastern North America. A key component of the beech bark disease complex, N. ditissima also causes disfiguring perennial cankers on numerous native hardwood species, with the greatest damage occurring on black and yellow birch, Betula lenta and B. alleghaniensis, respectively. With the goal of examining the relationship between mating system and genetic structure, a set of 13 polymorphic microsatellite markers were developed and used to genotype ascospore progeny from 44 perithecia isolated from two nearly adjacent sites at West Rock Ridge State Park in New Haven, CT. The 13 markers ranged in allelic diversity from 2-15 alleles/locus, with an average of 4.3 alleles per locus in this population. Pooled sibling ascospore haplotypes for each of the 44 perithecia were used to infer 27 unique multilocus diplotypes, from which parameters of population structure were estimated. Minimally significant (p≤0.05) linkage disequilibrium (LD) was observed between several pairs of loci, but a standardized estimate of multilocus LD was not significant. Genetic differentiation between the two subpopulations was minimally significant at three loci (Dest = 0.11, 0.17, 0.19), while the harmonic mean across all loci was not (Dest = 0.01); therefore, all subsequent analyses were performed on the entire population. Absence of segregation among sibling haplotypes was observed in 23 of the 44 perithecia, yielding a selfing rate, s, of 0.52. Heterozygote deficiencies were observed at all loci; Fis, the inbreeding coefficient, ranged from 0.39 to 1.0, and averaged 0.71 over all loci. Fis estimated from only the outcrossed portion of the population (n=21) was 0.42, supporting the hypothesis that biparental inbreeding contributed significantly to overall inbreeding in this population. These observations support theoretical models that posit the importance of biparental inbreeding to the evolutionary stability of mixed mating.

This abstract is not yet assigned to a session.

: MSA Student Workshop: Using fungi to get a job

Anne Pringle. Organismic and Evolutionary Biology, Harvard University, Cambridge, MA, 02138.

Some ads specifically ask for mycologists, more often, departments are looking for conceptually oriented biologists. Fungi offer myriad opportunities for connecting to broader themes in ecology, evolution, and other fields; making those connections can be critical to a successful job hunt. I'll talk about my own experiences on the job market and share the research and teaching statements I used to find my current position. We'll then practice job strategies by taking the time to talk about our research "mission statements" and "cocktail-party tidbits". I'll explain why these are useful and we'll finish by sharing our newly crafted mission statements and tidbits with each other.

Contributed Oral Session 1
Day: Monday, 16 July
Location:
Time: 0945

: Biofilm formation by Aspergillus niger, Aureobasidium pullulans, and Cladosporium cladosporioides

Julia Kerrigan* and Virginia Waldrop. School of Agricultural, Forest, and Environmental Sciences, 114 Long Hall, Clemson University, Clemson, SC 29634-0310.

A biofilm is an aggregate of microorganisms adhered together, to each other and to a substrate, surrounded by an extracellular matrix. Biofilms are ubiquitous, occurring in natural and artificial water systems, and are of great concern because of their persistence in industrial, medical, and household environments. Although bacterial biofilm research is well established, fungal biofilm research is comparatively new and the vast majority of the research has been conducted on yeasts. This research was undertaken to develop a repeatable system for studying biofilm formation by different types of filamentous fungi and to document their phenotypic changes throughout biofilm development. Aspergillus niger, Aureobasidium pullulans, and Cladosporium cladosporioides were studied because they are cosmopolitan, commonly found in biofilms, and have characteristic culture morphology allowing for easy detection of contaminants. We established a method for engineering biofilms in a controlled reactor that models those growing under low sheer. This method can be modified for different types of filamentous fungi and other microbes. The first stage of filamentous fungal biofilm formation involves spore attachment and germination. Hyphal tip growth produces an extracellular polysaccharide that helps the fungus grow and allows for attachment within the liquid environment. Hyphae form an interconnected network that attaches to itself and other surfaces. As hyphal proliferation continues, spores are produced, germinate, and give rise to additional biofilm mass. A mature biofilm is composed of a confluent mucilaginous hyphal matrix covered with conidia; however, the morphological changes during biofilm formation vary substantially among the species. The establishment of a repeatable filamentous fungus biofilm formation system and understanding the morphological steps involved in biofilm formation allow for future studies on how to prevent and remove them.

Symposium 1 - Phylogenetic, Ecological, and Functional Diversity of Fungi
Day: Monday, 16 July
Location:
Time: 1330

: Introduction to phylogenetic, ecological and functional diversity of fungi

Brendan P Hodkinson1 and Thomas Jarvie2*. 1International Plant Science Center, The New York Botanical Garden, 2900 Southern Blvd., Bronx, NY 10458-5126, 2International Products Manager, Roche 454 Life Sciences, 15 Commercial Street, Branford, CT 06405.

The representative from Roche/454 Life Sciences will give a 10-minute introduction to 454 sequencing, followed by a 5-minute introduction to the symposium by Brendan P. Hodkinson.

Contributed Oral Session 7
Day: Tuesday, 17 July
Location:
Time: 1345

: Phylogenetically marking the limits of the genus Fusarium for post-Article 59 usage

David M Geiser1*, Alejandro P Rooney2, Todd J Ward2, Takayuki Aoki3, Seogchan Kang1, Stephen A Rehner4, and Kerry O\\\'Donnell2. 1Department of Plant Pathology, Penn State University, University Park, PA 16802, 2USDA-ARS-NCAUR, 1815 N. University St., Peoria, IL 61604, 3MAFF, NIAS, 2–1–2, Kannondai, Tsukuba, Ibaraki 305–8602, Japan, 4Systematic Mycology and Microbiology Laboratory, USDA, ARS, Beltsville, MD 20705.

Fusarium (Hypocreales, Nectriaceae) is one of the most important and systematically challenging groups of mycotoxigenic, plant pathogenic and human pathogenic fungi. We conducted maximum likelihood (ML), maximum parsimony (MP) and Bayesian (B) analyses on partial nucleotide sequences of genes encoding the largest (RPB1) and second largest (RPB2) RNA polymerase II subunits on 94 fusaria to infer phylogenetic relationships within the genus and 20 of its near relatives. Our analysis revealed that Cylindrocarpon spp. formed a basal monophyletic sister to a 'terminal Fusarium clade' (TFC sensu Gräfenhan et al. 2011), which comprised 20 strongly informally named clades. Inclusion of the basal most divergences within the TFC received strong support only from the Bayesian analysis (posterior probability (B-PP) = 0.99-1), with clades including F. ventricosum and F. dimerum forming the two earliest diverging lineages. An internode supporting the remaining TFC, however, was strongly supported by MP and ML boostrapping (ML-BS 100%, MP-BS 87%) and a B-PP of 1. This contrasts with the published analysis of a phylogeny based on RPB2 and acyl citrate lyase gene sequences, which did not yield significant support for this clade. Diversification time estimates were made to infer the origins of the genus and its clades. Because 1) the TFC includes all economically important fusaria, 2) virtually all of its members produce a recognizable Fusarium anamorph, 3) the Fusarium anamorph names are widely used compared to the associated teleomorphs; applied mycologists rarely if ever see the latter, and 4) it corresponds most closely to a fairly stable concept of the genus that has existed for nearly a century, herein we formally combine several teleomorph genera within the TFC and their types into Fusarium. The present study provides a robust framework to guide future comparative phylogenetic and phylogenomic analyses, including work to test and extend the hypotheses developed herein.

Contributed Oral Session 4
Day: Tuesday, 17 July
Location:
Time: 0930

: Saturated molecular census of soil fungi in boreal Alaska suggests 6 million species of Fungi on Earth

Lee Taylor1*, Jack W McFarland2, Teresa N Hollingsworth3, and Roger W Ruess1. 1Institute of Arctic Biology, 311 Irving I Building, University of Alaska Fairbanks, Fairbanks, AK 99775, 2U.S. Geological Survey, 345 Middlefield Road, M.S. 962, Menlo Park, CA 94025, 3Boreal Ecology Cooperative Research Unit, PNW Research Station, USDA Forest Service, Fairbanks, AK 99775.

There are 98,000 described fungal species and the true number of fungal species is estimated to range from 0.6M based on a comparison of animal species to 1.5 M based on the number of pathogenic fungi associated with plants. Because systematics in fungi and other microbes is vastly different from that in animals, because most fungi are not plant pathogens, and because molecular methods were not used, existing estimates are likely to be very wrong. We used an environmental, molecular approach to census plant and fungal diversity in a simple ecosystem, with particular concern for adequacy of sampling and accuracy of species assessment. Our estimate, based on fungi found in soil associated with plants at a ratio of 17 fungi to 1 plant, is 6M fungal species, itself a conservative estimate. Our result indicates fungal diversity has been greatly underestimated. Our data offer an explanation for this underestimate; niche partitioning on a very fine scale. Furthermore, we find that species in the same genus and broad functional guild often occupy different niches. With the ongoing reorganization of biodiversity on Earth, it is critical to accelerate our documentation of the distribution and function of the likely 98% of fungal species that are currently undescribed.

Contributed Oral Session 13
Day: Wednesday, 18 July
Location:
Time: 0830

: Global population genetic structure of Aspergillus niger and Eurotium rubrum in green coffee beans

Iman A Sylvain* and Timothy Y James. University of Michigan, Department of Ecology and Evolutionary Biology. 830 North University Ann Arbor, MI 48109.

Whether small species are ubiquitous and cosmopolitan remains a significant question in mycological research. Whether "everything is everywhere," and how habitat properties or historical contingency influence the distribution of fungi has yet to be fully understood. Determining species ranges for fungi can be challenged by reliance on morphological characters for the identification of species, and enlisting molecular techniques has shown that fungi may have varied ranges that scale from largely global to narrowly endemic. Fungi associated with highly mobile agricultural products may have global distributions that mirror the trade of their plant hosts, or show patterns of biogeographic structure influenced by the ecology of distinct agroecosystems. Trans-global gene flow in fungi may be facilitated by the movement of agricultural commodities, resulting in panmictic populations, and the observation that fungi show no correspondence between genetic and geographic ranges. We are developing global coffee agriculture as a model system to understand how the movement of food products influences the dispersal of fungi. We have used a multi-locus sequencing technique to analyze the population structure of two mycotoxigenic Aspergillus species, A. niger and Eurotium rubrum, cultured from green coffee beans. Coffee beans were sourced from eight countries: Kenya, Ethiopia, Mexico, Costa Rica, Guatemala, Papa New Guinea, Bali, and Sumatra. The coffee was either produced as USDA-certified organic, or by conventional agricultural methods, and either wet or dry-processed. Phylogenetic trees constructed from rRNA ITS sequences suggest that multiple distinct, possibly cryptic, species are present within the sampled A. niger and E. rubrum clades. Beta-tubulin and calmodulin sequences show no geographic structure in these species, suggesting that Aspergillus species associated with coffee production may truly be everywhere.

Contributed Oral Session 8
Day: Tuesday, 17 July
Location:
Time: 1500

: Thelephoroid symbionts of Pisoniae are limited to a small group of closely related taxa throughout the range of the plant tribe

Jeremy Hayward* and Thomas R Horton. Department of Environmental and Forest Biology, Syracuse University, Syracuse, NY 13210.

While several studies have demonstrated greater ectomycorrhizal (ECM) specificity in the tropics than in the temperate zone, little is known about the patterns underlying this specificity. Here, we present preliminary results from a study investigating the structure of specificity between members of the Nyctaginaceae and the ECM /tomentella-thelephora lineage. Within the Nyctaginaceae ectomycorrhizal plants are restricted to the genera Neea, Pisonia and Guapira, all members of the tribe Pisoniae. Following a previous observation that two Pisoniae species in opposite hemispheres associated with closely related thelephoroids, we hypothesized that members of the Pisoniae, whatever their geographic context, would also associate with this clade. We sampled ectomycorrhizas from members of the Pisoniae growing in the Pacific and on the Caribbean islands of Dominica, Vieques and Puerto Rico. We used molecular barcoding to identify ECM fungi and analyzed their phylogenetic placement using Bayesian and Maximum-Likelihood techniques. We also undertook a literature review to identify fungi previously detected associating with members of the Pisoniae. We found that at the familial level, the ectomycorrhizal specificity of the Pisoniae is low: species associate with members of at least five ECM fungal families. In contrast to this pattern of family-level generalism, those thelephoroid ECM fungi associating with the Pisoniae cluster strongly within a single lineage, implying narrow specificity between the Pisoniae and their thelephoroid symbionts at a subgeneric level. This clustering pattern is well-supported in nonparametric tests of phylogenetic signal. This pattern of specificity holds true in the Pacific, in the Caribbean and in South America; it also holds true for representatives of all ectomycorrhizal plant genera within the Pisoniae. Based on these results, we hypothesize that phylogenetic placement may matter more than location in determining associations among tropical trees displaying ECM specificity. This hypothesis should be tested in associations between other tropical ECM plants and fungi.

Contributed Oral Session 3
Day: Monday, 16 July
Location:
Time: 0930

: Genetic diversity and reproductive isolation in Didymella bryoniae, a fungal pathogen of cucurbits

Marin T Brewer* and Ashley N Turner. Department of Plant Pathology, University of Georgia, Athens, GA 30602.

We are interested in determining the center of diversity, as well as historical and ongoing migration patterns in the cucurbit pathogen Didymella bryoniae. This fungus causes gummy stem blight and black rot of cucurbits everywhere they are grown, yet relatively little is known about how it is dispersed. Previous studies with dominant markers have indicated that this species is diverse and contains at least two distinct genetic groups. We sequenced four nuclear regions, including calmodulin (CAL), beta-tubulin (TUB2), chitin synthase 1 (CHS-1), and the internal transcribed spacer of the rDNA (ITS) of thirty-four isolates from diverse cucurbit hosts across the United States (US). Isolates from both genetic groups were represented in our sample. A total of 1415 bp were sequenced, resulting in 41 polymorphic sites and 13 unique haplotypes. A haplotype network constructed with TCS demonstrated that there are two distinct genetic groups, which is consistent with previous studies. However, a single isolate collected from a greenhouse in California was quite distinct from both groups. There is no association with genetic group and host species of origin, which is not surprising because earlier studies indicated that host specialization is not evident with D. bryoniae. Geographic associations were detected, however, as one of the groups was present only in the northeastern US whereas the other group was found throughout the US. There are no detectable morphological differences between the two genetic groups indicating that these groups represent cryptic species that are reproductively isolated. Studies on a worldwide collection of isolates are currently underway. Coalescent analyses on this collection will be aimed at understanding migration patterns, geographic origin, and approximate age of the divergence of the genetic groups to elucidate the evolutionary processes contributing to the observed patterns of diversity.

Contributed Oral Session 9
Day: Tuesday, 17 July
Location:
Time: 1430

: The biology and phylogeny of Rozella (phylum Cryptomycota) revealed by genome sequencing

Timothy Y James1*, Adrian Pelin2, Stefan Amyotte2, Courtney S Frye1, Nicolas Corradi2, and Jason E Stajich3. 1Dept. of Ecology and Evol. Biol., University of Michigan, Ann Arbor, MI 48109 USA, 2Dept. of Biology, University of Ottawa, Ottawa, ON, Canada K1N 6N5, 3Dept. of Plant Pathology and Microbiology, University of California, Riverside, CA 92521 USA.

The enigmatic genus Rozella is a chytrid-like endoparasite of water molds that is related to a diverse array of organisms (Cryptomycota) known only from environmental DNA sequences. Rozella and other Cryptomycota are suggested to lack a chitinous cell wall during at least part of their trophic phase. We produced a draft genome sequence for Rozella using a combination of next generation sequencing technologies. Rozella possesses at least 5 chitin synthase genes which it uses during both resting sporangium formation and host invasion. The mitochondrial genome of Rozella is reduced in size and gene content, displaying a genome remarkably convergent with that of Plasmodium. A phylogenomic analysis confirms the placement of Rozella on the primary (earliest diverging) branch of the fungal kingdom along with other Cryptomycota such as microsporidia. The nuclear genome, however, has not undergone the major reduction in coding potential as observed in microsporidia.

Symposium 2 - The science that underpins fungal conservation
Day: Monday, 16 July
Location:
Time: 1410

: Species concepts in European stipitate hydnoid fungi

A Martyn Ainsworth1*, David Parfitt2, Hilary J Rogers2, and Lynne Boddy2. 1Royal Botanic Gardens, Kew / Natural England, Mycology Section, Jodrell Lab., Kew. TW9 3AB. United Kingdom, 2Cardiff University, Cardiff School of Biosciences, Cardiff. CF10 3AX. United Kingdom.

The standard taxonomic treatment of European stipitate hydnoid species (ectomycorrhizal tooth fungi in the genera Bankera, Hydnellum, Phellodon and Sarcodon), written by R.A. Maas Geesteranus, is almost 40 years old. It is timely therefore to re-examine the morphological species concepts therein using the results of a combined molecular and morphological approach. These fungi have received conservation-related publicity across Europe and are now generally regarded as nitrogen-sensitive tree symbionts frequently associating with roots of Fagaceae and Pinaceae. A decline in European fruiting populations is mainly ascribed to increased habitat loss and aerial nitrogen deposition. Of the 18 extant stipitate hydnoid species currently included on the British and Irish checklist, 15 are recognised as of conservation importance (priority BAP species) in the UK. However all conservation status assessments currently depend on morphological taxonomic concepts and accurate identification of fruit bodies. Recognising that a mushroom is a stipitate hydnoid is often straightforward, but naming the species on existing fruit body criteria can be fraught with difficulty. This is due to a combination of identification and taxonomic issues. In order to improve assessments of species distributions, conservation status and ecological/conservation requirements, we have adopted a combined approach using molecular (ITS1 sequencing) and traditional (fruit body and spore morphology) methods. We have discovered that the list of extant taxa is rather longer (and increasing) than the list of currently accepted names. There are undoubtedly cryptic and not-so-cryptic hydnoid taxa to be described in Britain and elsewhere in Europe. We are now comparing species names with sequence-based groupings in order to pin-point the undescribed species. This combined approach will facilitate stipitate hydnoid identification, help to locate their "best" sites for conservation and accelerate their below-ground ecological study.

Contributed Oral Session 5
Day: Tuesday, 17 July
Location:
Time: 1000

: Studies in Geoglossomycetes systematics

Vincent P Hustad1,2* and Andrew N Miller2. 1Dept. of Plant Biology, University of Illinois, 505 South Goodwin Ave., Urbana, IL 61801, 2Illinois Natural History Survey, University of Illinois, 1816 South Oak St., Champaign, IL 61820.

Geoglossomycetes are a widespread and diverse class of fungi known from every continent except Antarctica. Commonly referred to as earth-tongues due to their morphology and terrestrial habitat, these fungi have long been a subject of mycological interest, although few molecular systematic studies have been conducted within the group. This research presents the latest results of our attempt to generate the first modern, multi-gene, species-level phylogeny within Geoglossomycetes. Sequences of the partial nuclear ribosomal 28S large subunit and internal transcribed spacer genes, as well as the protein-coding genes, RPB1 and MCM7, were generated for numerous species previously thought to belong in Geoglossomycetes. Results of maximum parsimony, maximum likelihood, and Bayesian analyses confirmed previous findings that Geoglossomycetes is a well-supported monophyletic class that includes the genera Geoglossum, Nothomitra, Sarcoleotia, and Trichoglossum. Polyphyletic genera within Geoglossomycetes were also observed and two new genera, Glutinoglossum and Sabuloglossum, were established.

Contributed Oral Session 11
Day: Tuesday, 17 July
Location:
Time: 1730

: Fungal gene clusters, transfer, tinkering and death

Jason C Slot*, Matthew A Campbell, Han Zhang, John G Gibbons, and Antonis Rokas. Department of Biological Sciences, Vanderbilt University, Nashville, TN.

The association of metabolically linked genes in chromosomal gene clusters has been shown to facilitate accelerated evolution of fungal genomes through whole pathway acquisition by horizontal gene transfer (HGT) and through accelerated pathway loss. How gene clusters originate and integrate into novel fungal genomes following HGT is not well understood, however. In order to address these questions, we investigated the evolution of several secondary metabolite gene clusters with highly restricted distributions among closely related fungal taxa. Here we present evidence that novel gene clusters can persist along with alternative assemblages of genes in polymorphic loci. These variable loci could explain the dramatic differences in secondary metabolic diversity between closely related fungi. Recently acquired clusters may alternatively be scrapped, with some components being repurposed and the rest left to decay. These results provide more insight into the importance of gene clusters in fungal metabolic diversity and suggest that the rate of gene cluster HGT is likely underestimated.

Symposium 4 - Compararive Genomics of Fungi
Day: Wednesday, 18 July
Location:
Time: 1330

: Insights into independent origins of multicellularity in Neolecta from comparative genomics

Jason E Stajich1*, David A Hewitt2, and Gregory Jedd3. 1Plant Pathology and Microbiology, University of California, Riverside, CA USA, 2Department of Botany, Academy of Natural Sciences of Philadelphia, Philadelphia, PA, 3Temasek Life Sciences Laboratory and Department of Biological Sciences, The National University of Singapore, Singapore.

The evolution of multicellular tissues in fungi, from the most fundamental aspects of hyphal growth to development of complex fruiting bodies, required a host of changes in cellular biology of fungi. Genome sequence comparisons provide a means to study the molecular evolutionary changes across species through the inventories of gene content and genome organization. Phylogenomic comparison of genes in the filamentous Pezizomycotina fungi reveals a collection of lineage specific genes including some known to be important for the filamentous lifestyle such as the Hex proteins of Woronin body organelles, which are necessary for septal pore plugging after hyphal wounding. Neolecta, a Taphrinamycotina fungus, is phylogenetically distinct from the Pezizomycotina and may represent an independent lineage where hyphal growth and complex fruiting bodies evolved. By sequencing and comparing the genome of Neolecta irregularis to the rest of the Ascomycete fungi we have found further evidence that filamentous-specific genes in Pezizomycotina remain specific to the clade. Microscopy reveals Wornin-body like structures in the hyphae of Neolecta, yet Woronin body genes hex, wsc, leashin and spa9 are not found, suggesting independent origins for this adaptation. The NADPH oxidases appear to be expressed in all multicellular fruiting bodies, and one of these is encoded in the Neolecta genome, suggesting shared traits that might have been present in a complex Dikarya ancestor. These and ongoing analyses will reveal further patterns of genome evolution that link cell biology changes to evolutionary transitions between unicellular and multicellular forms in fungal history.

Contributed Oral Session 15
Day: Wednesday, 18 July
Location:
Time: 0845

: The occurrence of Phialosimplex in the built environment

Joey B Tanney* and Keith A Seifert. Eastern Cereal and Oilseed Research Centre, 960 Carling Avenue, Ottawa, Ontario, K1A 0C6.

House dust samples were collected from 14 countries as part of an ongoing project comparing the indoor mycota detected by dilution-to-extinction culturing and pyrosequencing. A modified isolation method using microtubes and low water activity agar media was used to culture slow-growing and xerophilic fungi that would potentially be missed by classical methods, which favor fast-growing mesophilic molds. Strains identified as Phialosimplex comprised approximately 15% of identified cultures from house dust samples from Micronesia, 0.8% from Indonesia, and an as yet unquantified percentage of strains from Thailand. Phialosimplex is a recently described anamorphic genus within the Trichocomaceae. Morphologically, it is characterized by narrow phialides borne on vegetative hyphae or on short, unbranched conidiophores, with aseptate subglobose to ovoid conidia in long chains. There are three known species, P. caninus, P. chlamydosporus, both associated with disseminated mycoses in dogs, and P. sclerotialis, originally isolated from hay. In a recently published study, the latter species comprised 37% of fungi identified in a poultry facility using pyrosequencing. Preliminary ITS data suggest the presence of at least one novel Phialosimplex species among the house dust isolates. Our ITS sequences of Phialosimplex will be used to identify OTUs in the 454 pyrosequencing data generated from the same dust samples, allowing us to compare the sensitivity of the microbiological and molecular methods for detecting species of this genus. The frequent occurrence in house dust of Phialosimplex, a genus previously considered as an obscure agent of canine disease, should encourage further investigation into its significance in the built environment.

Symposium 7 - Using cellular structure and biochemistry as indicators of fungal phylogeny
Day: Wednesday, 18 July
Location:
Time: 1600

: The structure of the hyphal apex: Spitzenkörper or not?

Robert W Roberson. School of Life Sciences, Arizona State University, Tempe, AZ 85287.

The defining feature of filamentous fungi is the hypha. Hyphal growth has aided the Fungi to successfully utilize a wide range of ecological habitats and develop multiple lifestyles. Cellular and molecular studies of hyphal growth have placed great emphasis on the Spitzenkörper (Spk). The Spk is a dense, roughly spheroidal cluster of vesicles, cytoskeletal components and signaling proteins found at the tips of most growing hyphae. It plays crucial roles in optimizing hyphal extension rates and in determining patterns of growth and morphogenesis by acting as a dynamic organizing center in the reception of secretory vesicles and orchestrating their delivery to the apical membrane. The Spk appears to have evolved only in filamentous fungi where it is present in all members of the Basidiomycota and Ascomycota studied thus far. Interestingly, a structural equivalent of the Spk is not common or has not been observed in hyphae of the zygomycete fungi (e.g., Mucoromycotina), or many the other relatively early diverging lineages. As notable exceptions to this, Spk have been identified in hyphae of Basidiobolus sp. (a zygomycete of uncertain phylogeny) and Allomyces macrogynus (Blastocladiomycota). In fungi that lack a recognizable Spk, vesicle clusters are often present in the hyphal apex. Although the arrangement of these vesicles is less organized and complex, there are characteristic patterns that are recognized. For example, in Rhizopus species, and other members of the Mucoromycotina, a thin crescent-shaped band of closely packed vesicles is present just beneath the apical plasma membrane. Although there have been great advances in understanding the biology and distribution of the Spk, profound questions remain. In this presentation, light and electron microscopy data will used to review cytoplasmic features of the hyphal apex from diverse fungal groups in hopes to better understand hyphal cell biology and fungal phylogeny.

Contributed Oral Session 9
Day: Tuesday, 17 July
Location:
Time: 1445

: Molecular phylogeny of the Entomophthoromycota

Andrii P Gryganskyi1,5*, Richard A Humber2, Matthew E Smith3, Jolanta Miadlikovska1, Steven Wu1, Kerstin Voigt4, Iryna M Anishchenko5, Oleksandr V Savytskyi6, and Rytas Vilgalys1. 1Duke University, Department of Biology, Durham, NC 27708-90338, USA, 2USDA-ARS BioIPM Research, RW Holley Center for Agriculture & Health 538 Tower Road, Ithaca, NY 14853, USA, 3Department of Plant Pathology, University of Florida, Gainesville, FL 32611, USA, 4Friedrich-Schiller University of Jena, Institute of Microbiology, 25 Neugasse, Jena 07743, Germany, 5M. Kholodny Institute of Botany, National Academy of Sciences of Ukraine, 2 Tereshchenkivska St., Kyiv 01601, Ukraine, 6Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, 150 Acad. Zabolotnogo St., Kyiv 03680, Ukraine.

The Entomophthoromycota, a recently recognized new phylum, is a ubiquitous group of fungal pathogens of a wide variety of economically important insect pests, and other soil invertebrates. This group also includes a small number of parasites of reptiles, vertebrates (including humans), macromycetes, fern gametophytes, and desmid algae, as well as some saprobes. Here we report recent studies to resolve the phylogenetic relationships within the Entomophthoromycota and to place this group reliably among other basal fungal lineages. Bayesian Interference and Maximum Likelihood analyses of four loci (nuclear 18S and 28S rDNA, mitochondrial 16S, and the protein-coding RPB2) as well as non-molecular data consistently and unambiguously identify 31 taxa of Entomophthoromycota as a monophyletic group distinct from other Zygomycota and flagellate fungi. Using the constraints of our multi-gene dataset we constructed the most comprehensive rDNA phylogeny yet available for Entomophthoromycota. The taxa studied here belong to five distinct, well-supported lineages. The Basidiobolus clade is the earliest diverging lineage. The Conidiobolus lineage is a paraphyletic grade of trophically diverse species that include saprobes, insect pathogens, and facultative human pathogens. Three well supported lineages in the exclusively entomopathogenic Entomophthoraceae center around the genera Batkoa, Entomophthora and Zoophthora, although several genera within this latter crown clade are resolved as non-monophyletic. Ancestral state reconstruction suggests that the ancestor of all Entomophthoromycota was morphologically similar to species of Conidiobolus. Analyses using strict, relaxed, and local molecular clock models documented highly variable DNA substitution rates among lineages of Entomophthoromycota. Despite the complications caused by different rates of molecular evolution among lineages, our dating analysis indicates that the Entomophthoromycota originated 405±90 million years ago. We suggest that entomopathogenic lineages in Entomophthoraceae probably evolved from saprobic or facultatively pathogenic ancestors during (or shortly after) the evolutionary radiation of the arthropods.

Contributed Oral Session 11
Day: Tuesday, 17 July
Location:
Time: 1600

: Modelling transcriptional reaction norm evolution in the model yeast Saccharomyces cerevisiae

Andrea Hodgins-Davis1*, Aleksandra Adomas2, Daniel Rice3, and Jeffrey Townsend1. 1Department of Ecology and Evolutionary Biology, Yale University, New Haven, CT 06520, 2National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, 3Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, MA 02138.

Genetic variation for plastic phenotypes has the potential to contribute a large proportion of the phenotypic variation available to selection for novel adaptation. Models for linking population variation with plasticity make predictions about how selection may constrain the evolution of gene expression variation across transcriptional reaction norms. To empirically test these models, we have characterized population variation for transcriptional reaction norms in Saccharomyces cerevisiae for an ecologically relevant gradient of copper concentrations from starvation to toxicity. We find that although the vast majority of transcriptional variation is small in magnitude, not just some, but most genes demonstrate variable expression across environments or genetic backgrounds. Functionally, the most highly expressed genes defined three distinct cellular states across the copper reaction norm consistent with previous results characterizing eukaryotic responses to copper starvation, copper-replete fermentation, and copper overdose. These cellular states included direct transcriptional responses to intracellular levels of copper ions and diverse indirect metabolic adjustments to the consequences of changed copper levels. Indirect homeostatic changes of expression were more variable among genotypes in their direction of response than were the reaction norms of genes directly regulated by copper-binding transcription factors. To interpret this variability in transcriptional reaction norms in the context of the processes of natural selection and neutral drift, we account for variation in mutation and regulatory degree across the genome by parameterizing classic models of phenotypic evolution. Empirically estimating population genetic variance, mutational variance, and regulatory degree, we infer the strength of stabilizing selection operating on gene expression levels. We present evidence consistent with either weak or infrequent stabilizing selection on most gene expression phenotypes, and discuss the implications of the lack of constraint for the evolvability of transcriptional reaction norms.

Contributed Oral Session 9
Day: Tuesday, 17 July
Location:
Time: 1415

: Genome-enabled marker development for evolutionary studies of the insect pathogen Metarhizium

Ryan M Kepler1*, Jonathan Shao2, and Stephen A Rehner1. 1Systematic Mycology and Microbiology Laboratory, USDA, ARS, Beltsville, MD, 20705, 2Molecular Plant Pathology Laboratory, USDA, ARS, Beltsville, MD, 20705.

Genome sequence resources for fungi enable development of custom genetic markers scaled to specific evolutionary inferences that improve greatly on information derived from the more limited set of generic legacy genetic markers. Using recently available genome sequences for M. anisopliae and M. acridum, we have developed a suite of novel genetic markers for investigating cryptic speciation, phylogeography and population genetics within Metarhizium s.l. To exploit the inherent variability of nuclear intergenic (nucIGS) regions we have created a simple graphical approach to visually scan for variable nucIGS regions flanked by conserved gene sequences that can serve as anchors for robust primer development. Using pairwise alignments of selected genomic scaffolds from the two species created in the Mummer 3.0 genome alignment program, a simple script was developed that tabulates the number of SNPs via a scalable sliding window function, facilitating visualization of trends in nucleotide substitutions and similarity between syntenic genomic regions among pairs of sequences. We describe the development of novel markers developed using this approach and demonstrate their utility to infer cryptic phylogenetic partitions previously missed or poorly resolved by pre-existing phylogenetic markers. We demonstrate the superior performance of these new versus preexisting markers by comparison of their phylogenetic informativeness using PhyDesign and Genealogical Sorting Indices. In addition we: 1) summarize the numbers, types and genomic distribution of microsatellite markers and compare these to those developed by enrichment methods, and 2) discuss the mating locus and development of improved mating type assignment PCR assays. Through selected examples we illustrate the improvements in evolutionary inference achieved with these newly developed genetic markers.

Contributed Oral Session 3
Day: Monday, 16 July
Location:
Time: 0945

: Transposable element dynamics in ectomycorrhizal genomes: a close-up perspective

Jaqueline Hess1*, Inger Skrede2, and Anne Pringle1. 1Department of Organismic and Evolutionary Biology, Harvard University, 16 Divinity Ave., Cambridge, MA 02130, 2Microbial Evolution Research Group (MERG), Department of Biology, University of Oslo, P.O. Box 1066 Blindern, N-0316 Oslo, Norway.

Ectomycorrhizal (ECM) symbiosis between plants and fungi has evolved several times in the Fungal Kingdom. Ongoing genome sequencing of ectomycorrhizal (ECM) fungi is increasing the understanding of the genetic mechanisms central to the evolution of the symbiosis. Besides changes in gene content and gene family sizes, differences in genome architecture are also apparent, likely caused by changes in the ecology and population dynamics of ECM fungi. A common feature of ECM genomes sequenced to date is greatly elevated transposable element (TE) content: approximately 20% of the genome of Laccaria bicolor and 60% of the genome of Tuber melamsporum is made up of TEs. The genus Amanita encompasses an evolutionary transition from a free-living, saprotrophic to ECM niche and provides a powerful model for interrogating genome evolution across closely related, ecologically distinct species. To investigate changes in TE dynamics after evolution of the ECM niche we used next generation sequencing technologies to shotgun sequence the genomes of five species, covering the major clades of the genus Amanita (three ECMs and two saprotrophs) and the saprotrophic outgroup species Volvariella volvacea. We found the ECM genome sizes to be expanded, ranging from one and a half to nearly double the size compared to the saprotrophic genomes. Here, we will present our informatics approach for TE detection from short read data and fragmented assemblies and discuss the patterns of TE invasion in the different ECM clades of Amanita.

Contributed Oral Session 11
Day: Tuesday, 17 July
Location:
Time: 1615

: Genomic and transcriptomic analysis of the novel endophytic isolate E5202H for the identification of natural product pathways

Daniel J Spakowicz*, Jeffrey J Shaw, Rahul S Dalal, Gang Fang, Brian F Dunican, and Scott A Strobel. Dept of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06511.

Endophytes are a diverse niche of fungi that produce potentially useful secondary metabolites. Isolate E5202H is an endophyte that was isolated from the plant Guazuma ulmifolia (Sterculiaceae) in the Cerro Blanco dry forest of Ecuador in 2009. This isolate is likely a new species in the genus Massarina, with 86% similarity in its internal transcribed spacer rDNA sequence to the closest named organism in GenBank by Smith-Waterman alignment. In addition, E5202H produces a novel molecule with anti-oomycete activity, shown by mass spectrometry and NMR to be (2E,4E,6E)-6-methylocta-2,4,6-trienoic acid. Also, it produces a volatile hydrocarbon with potential biofuel properties, shown by mass spectrometry and NMR to be (3E,5E,7E)-nona-1,3,5,7-tetraene. Metabolic labeling studies suggest that (3E,5E,7E)-nona-1,3,5,7-tetraene is produced by a polyketide synthase-like mechanism. Genomic and transcriptomic analyses of E5202H are being pursued to identify the genes involved in the production of these molecules. The E5202H genome has been assembled to 20 scaffolds with an estimated size of 52 MB. We pursued transcriptome sequencing from five different conditions where different volatile metabolites were produced, as determined by GCMS. Correlating the transcription with metabolite production and isotopic labeling data allows us to generate hypotheses for the genes responsible for the production of these products. The functions of candidate genes will be determined by heterologous expression studies.

Symposium 1 - Phylogenetic, Ecological, and Functional Diversity of Fungi
Day: Monday, 16 July
Location:
Time: 1410

: High-throughput sequencing to explore the biogeography of soil fungi and the use of network analyses to examine fungal-bacterial interactions

Scott T Bates1*, J Gregory Caporaso2, D Lee Taylor3, and Noah Fierer1. 1Cooperative Institute for Research in Environmental Sciences, University of Colorado, Boulder, CO, USA, 2Department of Computer Science, Center for Microbial Genetics and Genomics,
Northern Arizona University, Flagstaff, AZ, USA, 3Institute of Arctic Biology and 
Department of Biology and Wildlife, 
University of Alaska Fairbanks, 
Fairbanks, AK, USA.

High-throughput DNA sequencing technologies have paved the way for new analytical approaches that move beyond basic descriptive inventories of microbial communities. With the aim of deciphering the structure of complex microbial communities across spatial or temporal gradients, network analysis allows for the examination of potential interactions between microbial taxa in natural systems using large environmental sequence datasets. Here, we sampled soils from numerous sites across North and South America, and used fungal ITS and bacterial 16S rRNA gene sequencing with Illumina technology to recover millions of environmental sequences from our sample sites. In addition to examining biogeography for soil fungi, we also applied network analysis approaches to identify significant co-occurrence patterns between fungi and bacteria on the basis OTU abundance and occupancy. We outline factors that influence broad-scale biogeographical patterns for fungi in terrestrial systems, and describe the topology of the resulting fungal/bacterial co-occurrence network. Finally, we discuss the utility of the network approach for exploring inter-taxon correlations to gain a more integrated understanding of microbial community structure and the ecological rules guiding community assembly.

Symposium 4 - Compararive Genomics of Fungi
Day: Wednesday, 18 July
Location:
Time: 1450

: Becoming A Fungus: Comparative Phylogenetic Studies of Evolution of Absorptive Nutrition

Mary L Berbee1*, Satoshi Sekimoto1, Ludovic LeRenard1, Joseph W Spatafora2, and AFTOL2 Working Group1. 1Dept. of Botany, University of British Columbia, Vancouver BC Canada, 2Dept. of Botany and Plant Pathology, Oregon State University, Corvallis OR USA.

Most fungi gain nutrients by secreting digestive enzymes from their hyphae into the surrounding matrix, and then absorbing the nutrients that diffuse back. We are exploring data from our community sequencing proposal to the US Joint Genome Institute to better understand the metabolic capabilities of early fungi. With new genome sequences from early-diverging fungi including aquatic chytrids and terrestrial zygomycetes, it becomes possible to compare secreted enzymes across phyla and to speculate on the evolutionary origins of their underlying genes. The enzymes that fungi secrete to break down cellulose or proteins have paralogs that are not secreted but function instead in housekeeping within the cell, in modifying polysaccharides or in recycling proteins. Duplication of the housekeeping genes, modification for secretion, and addition of domains for binding to particular substrates can result in enzymes adapted for extracellular function. The Ascomycota and Basidiomycota share secreted enzymes with functions including breaking down cellulose. The chytrids and zygomycetes also have the housekeeping homologs to the secreted enzymes. However, true orthologs to the secreted enzymes of the Ascomycota and Basidiomycota seem to be rare among the early diverging clades. Assuming that it was the accumulation of plant biomass on land that selected for the maintenance of the enzymes of the Ascomycota and Basidiomycota, the absence of conserved secreted enzymes across other phyla could be explained if other fungi diverged too early to encounter land plants as a readily available source of nutrients.

Contributed Oral Session 15
Day: Wednesday, 18 July
Location:
Time: 0900

: Like a rock?: lack of sequence variation in clinical isolates of the dermatophyte Trichophyton rubrum

Michael T Gajdeczka*, Wenjun Li, Catharine B Kappauf, and Joseph Heitman. Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, North Carolina, United States of America.

Trichophyton rubrum (Ascomycota) - the cause of athlete's foot, ringworm and nail infections - is the most common dermatophyte affecting humans. Despite availability of an annotated genome, this microbe's pathogenicity, epidemiology and genetics remain poorly understood due to an inability to conduct genetic crosses, lack of adequate sampling and apparent clonality. The only evidence of sexual mating in T.rubrum is indirect - a single genetic hybrid was formed between T.rubrum and Arthroderma simii, the teleomorph of a related species (T.mentagrophytes). T.rubrum lacks a known teleomorph, and although its genome contains mating type genes, only a single MAT allele (MAT1-1) is known. This study has three aims: (1) survey sequence variation in our 194 clinical samples using sequence-based and previously-developed variable number tandem repeat (VNTR) markers; (2) use paired-end Illumina sequencing to identify additional polymorphism within and among strains belonging to different VNTR types, and (3) search for evidence of (same-sex) mating using genetic and culture-based methods. First, we sequenced 7.5 kb of mainly non-coding regions located within 20 kb of the ABC1 and CAP59 genes in ten T.rubrum isolates collected over a span of ten years. These sequences revealed six singleton polymorphisms, which distinguished only half of the strains. We are now transitioning to VNTR genotyping and whole genome Illumina sequencing to characterize the diversity more completely. Lastly, in order to test for formation of ascomata (sexual structures), we will co-culture ten of our T.rubrum isolates and a T.tonsurans isolate (MAT1-2) in single, pair-wise and triple combinations on dermatophyte mating medium. Cultures with extensive hyphal coiling (indicative of early sexual development) will be dissected and examined for presence of ascomata or ascospores. Thus far, our results provide no evidence of sexual reproduction and re-confirm low sequence variation in T.rubrum clinical isolates.

This abstract is not yet assigned to a session.

: Evolutionary aspects of oligotrophism and extremotolerance in Ascomycota

Martin Grube1*, Cene Gostincar2, and Lucia Muggia1. 1Institute of Plant Sciences, Karl-Franzens-University Graz, Graz, Austria, 2Department of Biology, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia.

As widely distributed colonizers of rocks, black meristematic fungi endure temperature extremes, desiccation and usually a low supply of nutrients. Certain traits, such as phenotypic plasticity and melanized cell walls have been suggested as important factors of their polyextremotolerance. Some of their adaptive abilities, e.g. growth at elevated temperatures, may also be considered as health concerns, especially since black fungi can often establish themselves in man-made habitats. Facultative lichen-like associations of black fungi with algae could contribute to the persistence in poor natural habitats on rocks. We detected several cases of such associations. Phylogenetic analyses already suggested an ancestral proximity of oligotrophic black fungi and certain lineages of lichen-forming fungi. We included further lichenized and lichenicolous species in phylogenetic analyses which seem to link certain groups of lichens with black fungi. We believe that the upcoming information on genomes from black fungi and comparative genomics will provide us with new insights into the pre-adaptations for extremotolerance and oligotrophism of black fungal lineages. This knowledge may also be used to predict the future evolutionary potentials of certain fungal lineages.

Symposium 3 - The mycologist's guide to the new International Code of Nomenclature for algae, fungi, and plants
Day: Monday, 16 July
Location:
Time: 1640

: Basidiomycete taxonomy without dual nomenclature

M Catherine Aime. Louisiana State University Agricultural Center, Department of Plant Pathology and Crop Physiology, Baton Rouge, LA 70803.

The new International Code of Nomenclature for algae, fungi, and plants (Melbourne Code) discontinued the application of dual nomenclature in Fungi. Under earlier Codes Article 59 permitted more than one name for pleomorphic non-lichenized basidiomycetes. Effective immediately all names will compete for priority. As of January 1, 2013, publication of alternative names for species or genera will not be allowed. These changes affect basidiomycete taxonomy in several ways. To make one example, anamorphic yeast states occur in all three subphyla of Basidiomycota. Under previous versions of the Code these were treated separately from teleomorphic species and assigned to form genera principally based on carbon assimilation tests. Molecular data are now used to place these within a phylogenetic framework, highlighting the artificiality of the old system. For example, species of Sporobolomyces occur across most of the yeast-forming Pucciniomycotina classes and species of Rhodotorula can be found across both Ustilaginomycotina and Pucciniomycotina, although the type species for both genera are placed in Sporidiobolales. Thus restriction of generic names to include only those species within a monophyletic lineage will dictate the necessity of many name changes within these groups as extra-Sporobolomyces, -Rhodotorula and other anamorphic basidiomycete yeast species are integrated into a phylogenetic-based taxonomy. These and many other taxonomic challenges for the community will be discussed as they apply to different groups of impacted Basidiomycota.

Symposium 7 - Using cellular structure and biochemistry as indicators of fungal phylogeny
Day: Wednesday, 18 July
Location:
Time: 1700

: Lysine pathways and cell wall chemistry revealed the existence of two evolutionary lines within the Kingdom Fungi

Salomon Bartnicki-Garcia. Dept. Microbiology, CICESE, Ensenada Center for Research & Higher Education, Baja California, Mexico.

Henry Vogel's discovery (1960's) splitting fungi into two camps based on lysine biosynthesis provided the first and strongest piece of evidence for the existence of two evolutionary lines within Fungi. By synthesizing lysine via the common route characteristic of bacteria, algae and plants, diaminopimelic acid path (DAP), Oomycetes and Hypochytridomycetes proved to be distinct from most Fungi, namely Chytridiomycetes, Zygomycetes, Ascomycetes and Basidiomycetes which synthesize lysine via aminoadipic acid pathway (AAA), a unique route found almost exclusively in fungi. Concurrent studies on cell wall chemistry of various fungi revealed drastic differences that correlated with lysine pathways, thus DAP fungi became members of the Cellulosic line, and AAA fungi of the Chitinous line. Later, molecular phylogeny convincingly confirmed the evolutionary schism among organisms traditionally called fungi but it also placed Mycology in an existential dilemma: either preserve phylogenetic purity by removing a most important group of organisms, the cellulosic fungi, from the kingdom or continue maintaining both cellulosic and chitinous fungi within the kingdom boundaries. While the first alternative has become widely adopted, it has serious negative repercussions. The second alternative would seem more practical and beneficial to the science of Mycology and to other fields where cellulosic fungi are of great importance. It benefits nobody to exclude Oomycetes from the real world of fungi namely, publications, review articles, congresses, classrooms, textbooks, and all sorts of academic and research opportunities focused on Fungi. I advocate setting aside the issue of phylogenetic purity, and declare the Kingdom Fungi as made of two separate evolutionary lines each giving rise to individuals different in biochemical details but displaying common physiologic and morphologic features... and please let's not refer to a classic fungus of the genus Phytophthora as a "protista-like" organism.

Contributed Oral Session 9
Day: Tuesday, 17 July
Location:
Time: 1345

: Ribosomal RNA gene-based and multi-gene phylogenies of Smittium (Harpellales) and allies-toward unraveling relationships among early-diverging fungi

Yan Wang1*, Eric D Tretter1, Eric M Johnson1, Prasanna Kandel1, Robert W Lichtwardt2, and Merlin M White1. 1Department of Biological Sciences, Boise State University, 1910 University Drive, Boise, ID 83725, 2Department of Ecology & Evolutionary Biology, University of Kansas, 1200 Sunnyside Avenue, Lawrence, KS 66045.

Smittium is one of the oldest members of the Harpellales, a group commonly referred to as the gut fungi. Gut fungi are endosymbiotic microorganisms that live in the digestive tracts of various Arthropods, worldwide. During the 75 years since the first species, Smittium arvernense, was described, Smittium has grown to include 81 species. This genus has also helped to advance our understanding of the gut fungi, by serving as a "model" for laboratory studies of the fungal trichomycetes. Many isolates of Smittium have been used for ultrastructural, physiological, host feeding, serological, as well as isozyme, and now ongoing molecular systematic studies. Previous and current molecular studies have shown that Smittium is polyphyletic but with consistent separation of Smittium culisetae, one of the most common and widespread species, from the remainder of Smittium species. Morphological (zygospore and trichospore shape), molecular (18S and 28S rDNA), immunological, and isozyme evidence were used to establish a new genus Zancudomyces, and to accommodate Smittium culisetae. A multi-gene dataset, consisting of 18S and 28S rDNA, RPB1, RPB2, and MCM7 translated protein sequences for Smittium and related Harpellales (Austrosmittium, Coleopteromyces, Furculomyces, Pseudoharpella, Stachylina and Trichozygospora), was used for phylogenetic analyses and provided strong support at multiple levels in the trees generated. The clades and branches of the consensus tree were assessed relative to morphological traits for the taxa of interest, including thallus branching type, holdfast shape, trichospore or zygospore characters as an aid to inform the taxonomy and eventual systematic revisions and reclassification.

Contributed Oral Session 9
Day: Tuesday, 17 July
Location:
Time: 1400

: Utilizing new genes (MCM7 and TSR1) for the phylogenetic analysis of early-diverging fungi

Eric D Tretter*, Eric M Johnson, Yan Wang, Prasanna Kandel, and Merlin White. Boise State University, Dept. of Biological Sciences, 1910 University Drive, Boise, ID 83725.

The Kickxellomycotina is a group of early-diverging fungi with highly diverse morphologies and life histories that were previously part of the non-monophyletic Zygomycota phylum. Phylogenetic studies of the Kickxellomycotina have been limited by our current inability to culture many members of the Harpellales and all members of the Asellariales, and the tendency of some clades to suffer from severe long-branch attraction (particularly the Dimargaritales). This reduces our ability to sample taxa within each order, and makes it difficult to find congruent gene trees to produce concatenated multi-gene analyses. Traditional gene loci may also have severe problems with alignment that limit phylogenetic power (ITS), have problems with paralogs (beta-tubulin), or may prove very difficult to amplify for samples not taken from culture (RPB1, RPB2). In an attempt to address these problems and find additional genes to improve our resolution within this subphylum, we have investigated the use of the genes MCM7 and TSR1, which have been proven to be powerful for phylogenetic inference within the Dikarya and congruent with trees produced by phylogenomic analysis. We developed primers suitable for the amplification of MCM7 and TSR1, within three of the current orders of Kickxellomycotina (Harpellales, Dimargaritales, and Kickxellales) and four genera that may represent new orders. To determine if these genes reveal phylogenies congruent with the evolutionary history of the organisms, we compare the TSR1 and MCM7 phylogenies to an analysis based on ribosomal RNA genes, the most commonly used and accepted gene locus for these taxa. Finally, we report on the success of attempts to amplify species within other groups of early-diverging fungi, and provide suggestions as to which primers could potentially work with other clades.

Contributed Oral Session 9
Day: Tuesday, 17 July
Location:
Time: 1330

: Molecular-based phylogenetic placement of the Asellariales, an enigmatic order of arthropod gut endosymbionts

Eric M Johnson*, Eric D Tretter, Prasanna Kandel, and Merlin M White. Boise State University, Dept. of Biological Sciences, 1910 University Drive, Boise, ID 83725.

The order Asellariales is comprised of three genera, Asellaria, Orcheselleria, and Baltomyces, which are endosymbionts in the guts of isopods and springtails. Ultrastructural studies suggest placement of Asellariales in the subphylum Kickxellomycotina, and life history further suggests a relationship to the Harpellales, an order of insect gut endosymbionts. However, placement of the Asellariales at any level has eluded verification due to a lack of molecular data. Attempts to obtain gene sequences for any member of the Asellariales have been seriously hindered for a variety reasons, including sparse DNA samples due to the microscopic size of the specimens; lack of culturability; and contaminating DNA from the host animal as well as other fungi, bacteria, and even host food material in the gut. Here we report the successful amplification and sequencing of partial sequences of 5 genes (SSU and LSU rDNA, RPB1, RPB2, β-tubulin) for Asellaria ligiae, as well as a single gene (LSU rDNA) for Orchesellaria mauguoi. We also describe the development and use of custom primers to obtain the sequences, as well as the unusual nature of A. ligiae rDNA and challenges associated with it. Finally, we can place representatives of the Asellariales using Bayesian inference on the basis of molecular data from Asellaria and Orchesellaria.

Contributed Oral Session 4
Day: Tuesday, 17 July
Location:
Time: 0900

: Application of semantic technology to define names for fungi

Nathan J Wilson1*, Kathryn M Dunn2, Han Wang2, and Deborah L McGuinness2. 1Marine Biological Laboratory, Center for Library and Informatics, 7 MBL St., Woods Hole, MA 02543, 2Rensselaer Polytechnic Institute, 110 Eighth Street, Troy, NY USA 12180.

The need for well-defined, persistent descriptions of taxa that can be accurately interpreted by computers is becoming increasingly clear. The goal of this work is to develop named descriptions of Fungi that enable automated reasoning by computers. We encode these descriptions using the Web Ontology Language (OWL). The initial target audience is field mycologists using the Mushroom Observer website, who range from professional scientists to beginning mushroom enthusiasts. We describe our mycology ontology along with a transparent, community-based ontology evolution process. The ontology was designed to focus on properties that can be observed in the field, but the framework is proving to be suitable for microscopic, chemical, and genomic properties as well. Concrete examples are provided where field mycologists need names for groups of similar-looking Fungi that are known to belong to different species, and where our approach can significantly increase the precision of information recorded by the observer. Such a system is important for enabling the field mycologist to make more meaningful contributions to the modern scientific literature. In addition, the resulting ontology and descriptions provides a foundation for consistent, unambiguous, computational representations of Fungi. Finally, we expect that such a system will enable more people to become active field mycologists by providing a more robust way to document field observations and connect those observations with information about similar fungi.

Contributed Oral Session 15
Day: Wednesday, 18 July
Location:
Time: 0930

: Assessment of the saprotrophic ability of the White Nose Syndrome agent, Geomyces destructans

Hannah T Reynolds1*, M Kevin Keel2, and Hazel A Barton1. 1Department of Biology, University of Akron, Akron, OH 44325, 2Department of Population Health, University of Georgia, Athens, GA 30602.

White Nose Syndrome, an emerging disease of hibernating bats, has killed over 5.5 million bats in northeastern North America. Its rapid spread across the country, beginning in New York in 2006 and currently reaching as far south as Alabama, has led to numerous cave closures across the United States. The fungus Geomyces destructans was isolated and described from infected bats and was recently shown to be the causal agent of the disease. Geomyces, the anamorph of Pseudogymnoascus, consists of 11 species known predominately from cold soils. Understanding the natural history of Geomyces destructans, particularly its ecology in caves, is crucial for disease management. To investigate the saprotrophic potential of Geomyces destructans, we used a combination of enzyme bioassays and cave community ecology. First, we tested G. destructans and other Geomyces species for the ability to decompose several common substrates, including chitin, keratin, and cellulose. We also tested for the production of siderophores, iron-chelating molecules that allow survival in low iron conditions and are important virulence factors for several pathogens. For positive tests, we compared the enzyme activity of G. destructans with that of closely related species. Second, we tested the growth ability of G. destructans in sterilized and unsterilized soils and sediments collected from multiple locations in caves in Cumberland Gap National Historical Park. For each sample, the microbial communities and soil composition were analyzed to determine the abiotic and biotic factors that permit the survival and growth of Geomyces destructans in caves.

Contributed Oral Session 4
Day: Tuesday, 17 July
Location:
Time: 0845

: ITS and LSU automated classification: the fungal RDP naïve Bayesian classifier

Andrea Porras-Alfaro1,4*, Liu Kuan-Liang3, Zachary Gossage1, Lynnaun Johnson1, Tabitha Williams1, Gary Xie2, and Cheryl R Kuske2. 1Department of Biological Sciences, Western Illinois University, Illinois, USA, 2Los Alamos National Laboratory, Bioscience Division, Los Alamos, New Mexico, USA, 3Institute of Information Management, National Cheng Kung University, Tainan City, Taiwan, Republic of China, 4Department of Biology, University of New Mexico, Albuquerque, USA.

The introduction of next generation sequencing (NGS) in ecological studies has created a major revolution in fungal ecology. Analyses of large fungal datasets are common but currently limited by the lack of databases and reliable tools that will allow the classification of thousands of fungal sequences from environmental samples. The main objective of this project was to create curated databases for the Internal Transcribed Spacer (ITS) region and the large-subunit rRNA (LSU) gene and to evaluate their performance in the Ribosomal Database Project (RDP) naïve Bayesian classifier. We created and compared hand-curated LSU and ITS databases (more than 8 000 fungal sequences per database). Taxonomic gaps in fungal databases and challenges in the creation and maintenance of these databases will be discussed. When compared with the traditional BLASTN approach, the RDP fungal classifier was more rapid (>460-fold with our system) with similar or superior accuracy. Performance of fungal classifier at different taxonomic levels, sequence lengths and regions was also evaluated. Classification was more accurate with 400-bp sequence reads than with 100-bp reads and location of hyper variable regions played an important role in accuracy of sequence classification. The fungal classifier shows to be a highly effective tool to analyze large NGS datasets from environmental surveys, and to determine target regions in LSU and ITS sequences that will improve accuracy of taxonomic classification when short sequences are obtained. The LSU training set and tool are publicly available through the Ribosomal Database Project (http://rdp.cme.msu.edu/classifier/classifier.jsp)

Contributed Oral Session 8
Day: Tuesday, 17 July
Location:
Time: 1345

: On the role of ectomycorrhizal fungi in Pinaceae invasions

Thomas R Horton1*, Martin A Nuñez2, Yazmin Rivera1, G Amico2, R Dimarco2, N Barrios2, Jeremy Hayward1, and Dan Simberloff2. 1Department of Environmental and Foresty Biology, SUNY-ESF, Syracuse, NY 13210, 2Department of Ecology and Evolutionary Biology, University of Tennessee, Knoxville, TN 37919.

Establishment of Pinaceae in locations with no history of the trees requires the co-introduction of compatible ectomycorrhizal (EM) fungi, which are also required to facilitate invasions. Compatible fungi disperse from plantations through mycelial networks that grow from plantation edges (short distance), and spores dispersed by wind, water and mammals (potentially long distance). Invasion of Pinaceae from plantations into native forests has been limited on Puerto Rico (USA) and Isla Victoria (Argentina). The trees do not associate with native EM fungi to any great extent, if at all. On Puerto Rico most established pines are within decimeters of plantations, but some trees have established at 1000m. Root tips collected from established pines at 1000m from plantations yielded Rhizopogon spp., as well as Pisolithus tinctorius, Wilcoxina mikolae and a member of the Atheliaceae. This indicates mammalian (Rhizopogon) as well as other dispersal vectors (Pisolithus, Wilcoxina, Atheliaceae). Bioassays with dried soils collected in areas 1000m from plantations and without pines only yielded Rhizopogon. Dispersal of fungi may be facilitated by rats and mice introduced since the 1500s. In addition, tropical storms and anthropogenic activity may vector the fungi over long distances. On Isla Victoria establishment is largely restricted around plantations. We have not observed the rare native deer or any rodents in or near plantations. However, nonnative deer were first introduced with conifers about 100 years ago and pigs have become numerous since 2004. A bioassay with ponderosa pine and Douglas-fir using fecal pellets from the nonnative mammals collected away from plantations yielded Rhizopogon, Suillus, and Hebeloma mycorrhizae. Severe weather during the peak fruiting period is probably less important for spore dispersal on Isla Victoria than it is on Puerto Rico. We hypothesize the degree long-distance dispersal and accumulation of compatible propagules can impact the rate at which these invasions proceed.

Contributed Oral Session 13
Day: Wednesday, 18 July
Location:
Time: 0900

: Population dynamics of Monilinia vaccinii-corymbosi in blueberry fields throughout the United States

Kathleen M Burchhardt* and Marc A Cubeta. North Carolina State University, Department of Plant Pathology, Raleigh, NC 27695.

Mummy berry disease of blueberry (Vaccinium spp.) is caused by the ascomycete Monilinia vaccinii-corymbosi (Mvc). The fungus produces asexual and sexual spores that infect blueberry shoots and fruit. While spore dispersal and sexual reproduction can contribute to the spread and evolution of fungal populations, little is known about the population dynamics of Mvc. The primary objective of this study was to utilize microsatellite-based genetic markers to examine intraspecific genetic diversity, population structure, and gene flow among populations of Mvc throughout blueberry growing regions of the United States. In this study, 438 isolates of Mvc sampled from 15 blueberry fields in New York, New Jersey, Massachusetts, Georgia, Mississippi, Oregon, Washington, Michigan, and North Carolina were screened with 10 polymorphic microsatellite markers. Results based on analyzing a geographically diverse subsample of 55 isolates from six fields indicate high intraspecific genetic diversity. Forty-eight private alleles were detected across the 10 loci, providing evidence for population differentiation and restricted gene flow among fields. Results from population genetic analyses of a more comprehensive dataset of 438 isolates will be presented.

Symposium 2 - The science that underpins fungal conservation
Day: Monday, 16 July
Location:
Time: 1510

: Conservation of Fungi - threat status of fungi in New Zealand and globally

Peter K Buchanan, Peter R Johnston, and Mahajabeen Padamsee*. Landcare Research, Private Bag 92170, Auckland 1142, New Zealand.

Fungi have been included in threat status assessments of New Zealand's biota since 2002. Assessments have mainly addressed macrofungi, as well as obligate species of fungi on threatened plants. The initial inclusion of fungi, and subsequent need for reassessment of threat status, has generated new research initiatives and raised awareness of fungal conservation. Aspects of fungal biology such as ephemeral reproductive stages add difficulty to threat status assessments. Examples are presented of fungal species listed in the highest (Nationally Critical) and other threat categories. Over 1,000 species of fungi are listed as Data Deficient due to inadequate distribution data. Fungi have been included in a prioritization exercise spanning all New Zealand's threatened taxa that are in decline, to evaluate methodology, feasibility, and cost of long-term recovery plans. Using molecular techniques, new records of ectomycorrhizal fungi listed as Data Deficient have been discovered by comparing environmental sampling of ectomycorrhizas with DNA sourced from herbarium specimens. Responding to increasing global awareness of the need for fungal conservation, the International Society for Conservation of Fungi was formed in 2010, and five Fungal Specialist Groups have been established under IUCN.

Contributed Oral Session 8
Day: Tuesday, 17 July
Location:
Time: 1400

: Ectomycorrhizal ascomycetes from the Guiana Shield: Elaphomyces and Pseudotulostoma

Terry W Henkel1*, Michael A Castellano2, Hannah T Reynolds3, M Catherine Aime4, Steven L Miller5, and Matthew E Smith6. 1Department of Biological Sciences, Humboldt State University, Arcata, CA 95521, 2USDA, Forest Service, Northern Research Station, 3200 Jefferson Way, Corvallis, OR 97331, 3Department of Biology, University of Akron, Akron OH 44023, 4Department of Plant Pathology and Crop Physiology, Louisiana State University Agricultural Center, Baton Rouge, LA 70803, USA, 5Department of Botany, University of Wyoming, Laramie, WY 82071, 6Department of Plant Pathology, University of Florida, Gainesville FL 32611.

The hypogeous false truffle genus Elaphomyces Fries (Elaphomycetaceae, Eurotiales, Ascomycota) contains ectomycorrhizal (ECM) fungi with a gleba of powdery, dark ascospores, a thick peridium, and large ascomata relative to other members of the Eurotiales. Of the ~67 species of Elaphomyces currently described worldwide, many are known from north temperate forests with ECM host plants in the Pinaceae, Fagaceae, and Betulaceae. Nearly a century ago Cooke and Rodway provided the first reports of Elaphomyces from the Southern Hemisphere (Australia), but erroneously assigned European names to their specimens. Recent efforts by Castellano et al. have uncovered at least 16 new Elaphomyces species from Australia and New Zealand. While Corner and Hawker in 1955 were the first to publish new Elaphomyces species from the lowland tropics in association with Dipterocarpaceae in Singapore, few other tropical species have been described. The recently discovered Elaphomyces digitatus ined. and Elaphomyces compleximurus ined. from Guyana constitute the first records for the genus for the lowland South American tropics. These fungi occur in rainforests dominated by ECM trees of the genus Dicymbe (Fabaceae subfam. Caesalpinioideae) and constitute the first records of Elaphomyces in association with leguminous host plants. Pseudotulostoma volvata, a stipitate, volvate member of the Elaphomycetaceae, is common in the same forests. So far, these fungi are the only ECM ascomycetes known from this lowland tropical ecosystem, despite the fact that more than 200 species of ECM basidiomycetes are present. We will discuss the morphology and ecology of these taxa, the evolution of ascoma form in the Elaphomycetaceae, and "missing" Elaphomyces diversity uncovered by root-based sequencing studies.

Contributed Oral Session 8
Day: Tuesday, 17 July
Location:
Time: 1415

: Ectomycorrhizal fungi associated with Pakaraimaea dipterocarpacea, an endemic neotropical dipterocarp

Matthew E Smith1, Terry W Henkel2*, H David Clarke3, Alex K Fremier4, and Rytas Vilgalys5. 1Department of Plant Pathology, University of Florida, Gainesville FL 32611, 2Department of Biological Sciences, Humboldt State University, Arcata CA 95521, 3Department of Biology, University of North Carolina-Asheville, Asheville, NC 28804, 4Department of Fish and Wildlife Resources, University of Idaho, Moscow ID 83844, 5Department of Biology, Duke University, Durham NC 27708.

Ectomycorrhizal (ECM) plants and fungi can be well represented in certain tropical regions. For example, the predominantly paleotropical ECM Dipterocarpaceae is one of the most widespread and speciose tree families in Southeast Asia. Pakaraimaea dipterocarpacea is one of only two species of dipterocarp known from the western hemisphere, and is the only member of the monotypic Dipterocarpaceae subfam. Pakaraimoideae. The species was discovered in Guyana by Bassett Maguire of the New York Botanical Garden in the 1950's, and is endemic to the sandstone highlands of Guyana and Venezuela in dense stands of multi-stemmed trees. In 2006 Bernard Moyersoen showed that Pakaraimaea forms ectomycorrhizas and provided evidence for a vicariant distribution relative to its Old World relatives. Despite the unique phylogenetic position of P. dipterocarpacea within the Dipterocarpaceae and its unique geographical distribution, little is known about the ECM fungi associated with the species. In 2010 we sampled ECM fungi on roots of P. dipterocarpacea and the leguminous ECM tree Dicymbe jenmanii where these species co-occur in the Upper Mazaruni River Basin of Guyana. Using molecular methods we documented 52 species of ECM fungi across 11 independent lineages. While many of ECM fungi found on Pakaraimaea roots were conspecific with taxa known from the region as sporocarps, a number of "unknowns" were found across several basidiomycete families and genera. Due to the phylogenetic distance between host taxa we hypothesized that P. dipterocarpacea would host ECM fungi not found on D. jenmanii, and vice versa. While statistical tests suggested that several ECM fungal species exhibited preference for either P. dipterocarpacea or D. jenmanii, most were generalists. In addition to these findings, unique ECM fungi collected as sporocarps in the Pakaraimaea forests will be discussed.

Symposium 2 - The science that underpins fungal conservation
Day: Monday, 16 July
Location:
Time: 1350

: Is there a "best" unit of conservation for valuable mushroom species? A scientific approach based on studies on matsutakes, morels and truffles

Matteo M Garbelotto* and Todd W Osmundson. Department of Environmental Science, Policy and Management, University of California, Berkeley, CA 94720, USA.

In the presence of limited economic resources, an approach is needed to maximize the results of conservation efforts focused on valuable mushroom species. Although the genomic nature of desirable mushroom phenotypes is still mostly unknown, we propose that a biogeographical approach combined with population genetics studies provide a concrete way to enact mushroom conservation plans. In the absence of precise knowledge of why a specific population should be protected, we suggest that the unit of conservation should be that of the "metapopulation" within each broad geographic region. It was first suggested in the late 1990s that habitat fragmentation was a major driver of speciation in Matsutakes. That intuition was recently confirmed by formal analysis on the effects of forest fragmentation on fungal community structure. The logistical advantage of focusing on habitat conservation, rather than on individual fungal species, is obvious. We show here that for recently evolved species, distribution of species tracks the presence of continuous forest cover, and an "isolation-by-distance" approach can be used to identify valuable conservation units. However, we present four examples of exceptions that need to be taken into account: a) The presence of significant geographic barriers, such as high mountain ranges, may accelerate drift; b) Significant bottlenecks caused by geologically recent events like glaciations may have led to genetic isolation between populations surviving in different refugia; c) Natural ecological events such as fires may be triggers of speciation and of hybridization among mushrooms; and d) Forest management may accelerate genetic drift by favoring short rotations where gene flow among populations is hindered.

Contributed Oral Session 17
Day: Wednesday, 18 July
Location:
Time: 1130

: Soil fungal lineages respond to host plant identity and community diversity in a model grassland system

Nicholas LeBlanc*, Linda Kinkel, and H Corby Kistler. Department of Plant Pathology, University of Minnesota, 495 Borlaug Hall 1991 Upper Buford Circle, St. Paul, MN 55108.

Grassland soils are known to harbor diverse fungal communities but the effects of host plant identity and the diversity of the surrounding plant community on these fungi is unknown. We have focused on rhizosphere fungal communities in a model grassland system using targeted metagenomics and well-annotated reference material for taxonomic assignment. A total of twenty-two samples were analyzed from hosts grown in monoculture or polyculture. Hosts included Big Bluestem, Little Bluestem, Round-Headed Bush Clover, and Sundial Lupine. We hypothesized that host identity and plant community diversity (monoculture and polyculture) would influence fungal community composition. Amplicon libraries targeting the LSU and RPB2 locus, derived from rhizosphere DNA, were sequenced using 454 technology. Sequences were aligned to custom databases composed of unique accessions published in phylogenetic studies, derived from genome databases, and obtained from the AFTOL consortium. Initial NCBI taxon IDs were used to assign reads to higher taxonomies using Perl scripts. Approximately 800 genera were detected through the LSU locus. Many of these groups represent slow-growing, poorly- studied fungi, such as Chaetothyriales and Verrucariales, which positively correlate with grass abundance. Other taxa, like Helotiales, were preferentially enriched in more diverse plant communities but also significantly correlated with soil potassium. The RPB2 locus revealed multiple Fusarium taxa with a predominance of F. oxysporum, for which host specific responses are suggested in certain groups. This work highlights the first steps towards linking plant host and community diversity with the abundance of root-associated fungi that likely play important, but poorly understood roles in plant growth and development.

Contributed Oral Session 4
Day: Tuesday, 17 July
Location:
Time: 1000

: A study of correlations between fungal and bacterial communities in dead wood

Björn Hoppe1*, Tiemo Kahl2, François Buscot1, and Dirk Krüger1. 1UFZ - Helmholtz Centre for Environmental Research, Department of Soil Ecology, Th.-Lieser-Str. 4, D-06120 Halle/Saale, Germany, 2University of Freiburg, Institute of Silviculture, Tennenbacherstr. 4, D-79085 Freiburg i. Brsg., Germany.

Dead wood (a.k.a. coarse woody debris, CWD) is a major habitat in forest ecosystems that is home and food for various types of organisms. Usually, researchers study the fungi and xylobiontic insects present and active in the wood decay process. The role of bacteria in the decomposition of this complex substrate is yet underexplored. One aspect of the project FunWood of the DFG (German Science Foundation) Biodiversity Exploratories is to investigate interrelations between fungal and bacterial diversity on CWD of European beech, Norway spruce and Scots pine along a gradient of forest sites that were under different silvicultural management practices. The same DNA extractions from drilled CWD served as templates for PCR subsequently processed for functional and diversity fingerprints (nitrogenase nifH T-RFLP, ARISA) as well as Sanger sequencing. Preliminary results entail 16S rDNA sequences from highly diverse eubacterial communities in CWD comprising 10 different classes. The detected families Beijerinckiaceae, Rhizobiaceae and Bradyrhizobiaceae are known to include potent nitrogen fixing bacteria. Conceivably, the extreme C/N ratios of CWD reaching 350-800:1 make it increasingly harder for wood decay fungi to meet their nitrogen demands as the wood decay process goes on. Thus, we aim to test further for the presence of actually nitrogen fixing bacteria and their potential role in filling the fungal nitrogen needs in this habitat. We hitherto found 26 different nifH sequences from 5 logs. None was closer than 97% similarity to any on GenBank, indicating the presence of uncharacterized, potentially unknown bacteria. By the time of presentation, statistically analyzed nifH sequence data of 48 CWD logs are ready to be discussed, along with fungal diversity data based on sporocarps and DNA.

Symposium 1 - Phylogenetic, Ecological, and Functional Diversity of Fungi
Day: Monday, 16 July
Location:
Time: 1430

: Diversity and phylogeny of marine and freshwater Verrucariaceae

Starri Heiðmarsson1*, Sergio Perez-Ortega2, Holger Thüs3, Cecile Gueidan3, Asuncíon de los Ríos2, and Francois Lutzoni4. 1Icelandic Institute of Natural History, Borgir Nordurslod, IS-600 Akureyri, Iceland, 2Departamento de Biología Ambiental, Museo Nacional de Ciencias Naturales, Madrid, Spain, 3Botany, Natural History Museum, London, United Kingdom, 4Biology, Duke University, Durham, United States.

Verrucariaceae consists mainly of lichenized members, although the lichenicolous lifestyle can also be found among members of this family. The family consists of crustose, squamulose, foliose and even subfruticose taxa. Within Verrucaria s. lat., delimitations of taxa has been problematic because only few phenotypic characters have been useful to infer relationships. Recent DNA-based phylogenetic analyses across this family have already resulted in the description of a genus comprising only marine members of Verrucaria s. lat. (Wahlenbergiella) and another genus which contains taxa that either grow by freshwater or in marine habitat (Hydropunctaria). Recently it was shown that Mastodia, which forms symbiosis with a leafy green alga, is part of the Wahlenbergiella clade. We will present the diversity of aquatic Verrucaria s. Lat. in Iceland, from both marine and freshwater habitats. Phylogenetic analyses of the genera Hydropunctaria and Wahlenbergiella were conducted, mainly based on Icelandic material, within a broader phylogenetic framework. These analyses revealed that the strictly marine clade Wahlenbergiella consists of at least two, fairly distinct clades. One of these clades comprises members of Wahlenbergiella, such as W. mucosa, W. striatula, W. Tavaresiae, and Verrucaria ceuthocarpa. The other clade consists of Mastodia tesselata and several taxa, which have been only collected in the southern hemisphere, so far.

Contributed Oral Session 10
Day: Tuesday, 17 July
Location:
Time: 1700

: Characterization of the wood decay fungi Fomes fasciatus and F. fomentarius using sequence analysis, morphology, and growth response to temperature in vitro

Meghan A McCormick*, Larry F Grand, and Marc A Cubeta. North Carolina State University, Department of Plant Pathology, Box 7567, Raleigh, NC 27695.

The wood decay fungi Fomes fasciatus and F. fomentarius are endemic to the United States. Both fungi cause a white heart rot of multiple species of trees and have essential roles in forest ecology and management. Little is known about the genotypic and phenotypic diversity of these species. For this study, basidiocarps of F. fasciatus and F. fomentarius were collected from 13 states within the United States. Pure cultures of each species were isolated from basidiocarp context tissue and/or single basidiospores. Ten morphological characteristics were recorded for basidiocarps and pure cultures. F. fasciatus and F. fomentarius were morphologically similar, but could be differentiated based on the smaller average basidiospore size of F. fasciatus. The average colony diameter (mm) was measured every 3 d for 9 d using five isolates each of F. fasciatus and F. fomentarius on malt extract agar at temperatures ranging from 12 to 36 °C in 4 °C increments. The temperature optima of F. fasciatus and F. fomentarius were 32 °C and 28 °C, respectively. In a separate study, we found that F. fasciatus grew at 39 °C while F. fomentarius did not. The genetic relatedness of the two species was examined using maximum parsimony analysis of the internal transcribed spacer region (ITS) of the ribosomal DNA (rDNA) and the RNA polymerase II gene (RPB2). Preliminary results based on 100 replications indicate that the two species represent distinct lineages that form two distinct clades with 100 percent bootstrap support. Further analysis of ITS rDNA and RPB2 sequences will be discussed.

Contributed Oral Session 5
Day: Tuesday, 17 July
Location:
Time: 1015

: A multigene phylogenetic assessment of species boundaries in Otidea

Karen Hansen* and Ibai Olariaga. Swedish Museum of Natural History, Department of Cryptogamic Botany, P.O. Box 50007, SE-104 05 Stockholm, Sweden.

Species of Otidea are among the more conspicuous Pyronemataceae (Pezizomycetes), producing large, typically ear-shaped apothecia. Otidea is restricted to the Northern Hemisphere, with the highest diversity in boreal-temperate forests, and is ectomycorrhizal. The genus is monophyletic and easy to recognize, but species delimitation is highly controversial. To resolve species boundaries and relationships within Otidea, we provide a multigene (LSU rDNA, RPB1, RPB2 and EF1; 4908 bp), worldwide phylogeny of Otidea and comparative morphological data. Two main lineages are supported within Otidea: I) O. alutacea, O. apophysata, O. daliensis, O. platyspora, and O. subterranea, and II) the rest of Otidea, which encompasses several sub-lineages. Otidea papillata and a clade of O. leporina and Otidea sp., are identified as successive sister lineages to the rest of lineage II. No unique morphological characters distinguish the two main lineages. The sub-lineages are supported by a combination of morphological characters, such as the apothecia shape and color, spore shape and ornamentation, paraphyses shape and content, and outer excipulum features. Our results, including sequences of type collections, show clear species limits, except within two species complexes: the O. alutacea-cochleata and O. phlebophora-rainierensis complexes. A number of Otidea names currently in use are shown to be synonyms. Several names have been interpreted differently in Europe, North America and Asia. At the same time some species seem to show continental endemism. Five new species have been discovered. The ITS region is too variable to align across Otidea, but appears to be useful for delimiting species; the ITS region is highly variable between species, but conserved within. To delimit species within the two species complexes we will perform separate analyses of these, including also the ITS region, and explore various analytical approaches for independent corroboration of the species identified. We estimate that Otidea comprises at least 35 species.

Symposium 5 - The Fungal DNA Barcode and Beyond
Day: Wednesday, 18 July
Location:
Time: 1350

: Assigning ITS and LSU rDNA 'barcodes' with confidence

Teresita M Porter* and G Brian Golding. McMaster University, Biology Department, Life Sciences Building, 1280 Main Street West, Hamilton, ON L8S 4K1 CANADA.

Whereas PCR, cloning and Sanger-based sequencing can generate libraries comprised of thousands of sequences up to about 600-1000 bp in length; current highly parallel sequencers are now generating amplicon libraries comprised of millions of relatively short sequences of about 200-400 bp in length. We are interested in how these shorter sequences affect the quality of automated taxonomic assignments. Here we present the results from two simulation studies that specifically address the accuracy and error rate of ITS and LSU rDNA markers using several tools suitable for automating assignments. We compare the performance of programs using similarity-, phylogeny-, and composition-based methods. Generally, we find that database completeness is perhaps the biggest factor determining assignment accuracy, followed by query sequence length, assignment method, primer choice, and sequence error. Specifically, we find that MEGAN lowest common ancestor (LCA) parsing produces the lowest error rate when assigning unknown ITS and LSU rDNA sequences; and, the fungal LSU naïve Bayesian classifier available through the Ribosomal Database Project website performs significantly faster than BLAST-based methods.

Contributed Oral Session 8
Day: Tuesday, 17 July
Location:
Time: 1515

: Community composition of ectomycorrhizal fungi along a climatic gradient in Scotland

Susan G Jarvis1,2*, Andy FS Taylor1,2, Ian J Alexander1, and Steve Woodward1. 1School of Biological Sciences, University of Aberdeen, Aberdeen, AB24 3UU, UK, 2The James Hutton Institute, Craigiebuckler, Aberdeen, AB15 8QH, UK.

Ectomycorrhizal (ECM) fungi play fundamental roles in forest ecosystems through tree nutrition and nutrient cycling. Understanding the responses of ECM fungi to environmental change will be important to maintain forest productivity for the future. Assessing the distribution of ECM communities along existing large-scale environmental gradients provides an insight into the environmental factors driving community composition. Previous studies have suggested nitrogen deposition to be the main environmental driver of ECM community variation over large-scale gradients in Europe. However, these studies have included areas with very high nitrogen deposition, which can dramatically alter ECM communities, and this may have masked any climatic effect on these communities. In Scotland nitrogen deposition varies with latitude but is less severe than in continental Europe, with total nitrogen deposition below 20 kg N ha-1 yr-1 in most areas. Utilising the latitudinal gradient in nitrogen deposition and a longitudinal climate gradient a survey of the ECM fungi present on the roots of native Scots pine (Pinus sylvestris) across Scotland was conducted to assess the relative impacts of climate and moderate nitrogen deposition on ECM community composition. Ectomycorrhizal pine roots were collected from 15 native pinewoods and fungal species were identified from root tips by sequencing of the ITS rDNA region. Non-metric multidimensional scaling of community composition and vector fitting of 17 climatic, nitrogen and soil chemistry variables identified temperature and rainfall variables to be correlated with the primary axis of community variation. Nitrogen deposition was shown to have little influence on community composition, although there was some effect of total soil nitrogen. This work highlights the potential for climate to influence ECM fungal communities and the need to understand the impacts, and possible interactions, of multiple drivers on community composition.

Symposium 5 - The Fungal DNA Barcode and Beyond
Day: Wednesday, 18 July
Location:
Time: 1410

: Oomycete barcoding and applications

C Andre Levesque*, Gregg Robideau, Chen Wen, and Lewis Christopher. Agriculture & Agri-Food Canada, Ottawa, Canada.

The ITS region is the de facto DNA barcode in oomycetes as in true Fungi. However, the cytochrome oxidase 1 (COI) region is the NCBI accepted barcode region and it must be ruled out to pave the way for an alternative marker such as ITS. COI does not work for the true Fungi, primarily because of numerous introns, but in oomycetes COI works at least as well as ITS. The prevalence of ITS in mycology has led to the proposal that it be included as barcode in addition to COI for oomycetes. Recently, the oomycete COI database was expanded to have comparable coverage of oomycetes species to the existing ITS data. COI has the advantage of being easy to align across all oomycetes because there are no introns and indels, whereas it is impossible to align ITS between genera. However, ITS was found to be a more valuable target in a DNA array system developed for most Phytophthora species. Specificity of hybridization probes is increased when they fall within an insertion or span deletions in closely related species. On the other hand, changes at the synonymous third codon positions are not as effective as they often yield very few base pair differences between closely related species over short oligonucleotides. Next generation sequencing provides additional applications of barcode sequence databases. The most common universal primers for ITS do amplify oomycete DNA and generate oomycete sequences in the large pool of PCR products processed by pyrosequencing. However, when studying ecology of oomycetes, the use of oomycete specific primers provides a better profile of species that are present at low propagule concentration.

Contributed Oral Session 13
Day: Wednesday, 18 July
Location:
Time: 0945

: Using new techniques in genotyping by sequencing to rejuvenate population genetic studies in fungi.

Andrew W Wilson1*, Norman Wickett1, Paul Grabowski2, Jeremie Fant1, Justin Borevitz2,3, and Gregory M Mueller1. 1Chicago Botanic Garden, Plant Conservation Science, 1000 Lake Cook Rd, Glencoe, IL 60022, 2University of Chicago, Ecology and Evolution, 1101 E 57th Street, Chicago, IL 60637, 3Australian National University, Research School of Biology, Canberra, Australia.

Challenges in obtaining sufficient numbers of informative population level markers has been an impediment to understanding the population genetics of fungi. Recent developments in high-throughput technologies have led to the innovation of new methods in genotype by sequencing (GBS) analysis. Restriction site associated DNA (RAD) sequencing is a relatively new technology that allows for identification of single-nucleotide polymorphisms (SNPs) by sequencing many small fragments in highly variable regions of the genome resulting from a restriction digest. Restriction sites are fairly conserved within a species, and RAD has been demonstrated as an effective way to generate loci from which SNPs can be identified for population genetic analysis. We tested the effectiveness of a novel GBS approach on 18 samples of the ectomycorrhizal species Laccaria bicolor. Initial data collection (one multiplexed lane of paired-end HiSeq 2000) produced over 64 million 101 bp reads. Preliminary analysis of all samples identified 13,528 SNPs consisting of 1919 multi-allelic loci. A comparison of Illinois (3 samples) and Wisconsin (2 samples) populations found 268 heterozygous loci, which produced an FST of 0.3229. Despite the small sample size, these preliminary results demonstrate the ability of the GBS approach to produce the data necessary for studying Agaricomycete populations. The results of a comprehensive analysis of the data generated in this pilot study as well as further discussion on the potential of these methods for fungal population genetic research will be presented.

Symposium 7 - Using cellular structure and biochemistry as indicators of fungal phylogeny
Day: Wednesday, 18 July
Location:
Time: 1640

: Using cellular structure and biochemistry for insights into gungal cell biology and phylogeny

Martha J Powell* and Peter M Letcher. Department of Biological Sciences, The University of Alabama, Tuscaloosa, AL 35487.

Chytridiomycota (chytrids) are now recognized as highly adaptable fungi, not limited in geography or habitat by the constraints of reproduction with a flagellated, unwalled-spore (=zoospore). As our examination of chytrid zoospore ultrastructure has expanded, we have discovered tremendous variety in organellar structures and organization. Molecular-based reconstructions of phylogeny have also revealed great genetic diversity among chytrids. The intersection of molecular phylogenies with analyses of organellar function is helping us understand how zoospore structures have evolved within lineages. Zoospore architectural differences are good predictors of molecular-based phylogenies. Selective pressures associated with habitat differences may be drivers in the evolution of zoospore ultrastructure. Evolution of zoospore architecture in the Chytridiales is discussed as an example of how structures have been modified and lost within evolutionary lineages. Our challenge now is discovering more about the functional roles of architectural design differences in chytrid zoospores.

Symposium 6 - Fungi in a Changing World
Day: Wednesday, 18 July
Location:
Time: 1600

: Temporal patterns of fungal fruiting reveal ongoing climate change effects

Håvard Kauserud1*, Einar Heegaard2, Ulf Büntgen3, Simon Egli3, Rune Halvorsen4, and Lynne Boddy5. 1Microbial Evolution Research Group (MERG), Department of Biology, University of Oslo, P.O. Box 1066 Blindern, NO-0316 Oslo, Norway, 2Norwegian Forest and Landscape Institute, Fanaflaten 4, N-5244 Fana, Norway, 3Swiss Federal Research Institute for Forest Snow and Landscape (WSL), Birmensdorf, Switzerland, 4Department of Research and Collections, Natural History Museum, University of Oslo, P.O. Box 1172 Blindern, NO-0318 Oslo, Norway, 5Cardiff School of Biosciences, Biomedical Building, Museum Avenue, Cardiff CF10 3AX, UK.

Effects of climate change on fungi may manifest in various ways. Time of fruiting may change, yields of fruit bodies can be altered and distributional ranges of species may shift. We are studying these three aspects using statistical analyses of time series data and digitized field and herbarium records. We show that the annual fruiting period across Europe has changed during the last 50 years. In Austria, Norway, Switzerland and the UK, the average annual fruiting time of autumn fruiters has been delayed. Moreover, the start and end of the annual fruiting period, using the 5 and 95 percentiles as proxies, have changed towards earlier and later, respectively. Changes in fruiting time have been especially dramatic in the UK, which deviates from the other regions by a more oceanic climate. Time series data from a Swiss forest plot indicates that the annual yields of ectomycorrhizal fruit bodies has increased dramatically during the last 40 years. We hypothesize that changes in the regional climate, including higher temperatures and more precipitation, can be linked to the increased fruit body yields. In another ongoing study we are exploring the extent to which fungi in the UK have shifted distributional ranges during the last 50 years and whether this can be linked to climate change.

Contributed Oral Session 7
Day: Tuesday, 17 July
Location:
Time: 1330

: Novel molecular markers and their utility in molecular systematics of Fungi

Jolanta Miadlikowska1*, Bernie Ball1, Francesc López-Giráldez2, Jeffrey P Townsend2, Ester Gaya1, Tami McDonald1, Suzanne Joneson1, Andrii Gryganskyi1, Teresita M Porter1, Brandon Matheny3, Kassian Kobert4, Alexandros Stamatakis4, Barbara Robbertse5, Joseph Spatafora5, David Hibbett6, Rytas Vilgalys1, and François Lutzoni1. 1Department of Biology, Duke University, Durham, NC 27708, 2Department of Ecology and Evolutionary Biology, Yale University, New Haven, CT 06520, 3Department of Ecology and Evolutionary Biology, University of Tennessee, Knoxville, TN 37996, 4Heidelberg Institute for Theoretical Studies, Schloss-Wolfsbrunnenweg 35, D-69118 Heidelberg, Germany, 5Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331, 6Department of Biology, Clark University, Worcester, MA 01610.

Although next generation sequencing methods have proven to be very successful in accelerating data acquisition, selecting the optimal set of molecular markers for phylogenetic studies has remained a complex endeavor. Existing fungal phylogenies demonstrate the urgent need for novel single-copy protein-coding genes to resolve phylogenetic relationships among fungi at all taxonomic levels with high confidence. As part of the Assembling the Fungal Tree of Life project (AFToL 2), a comparative genomic approach was adopted to select all single-copy orthologous genes with the greatest potential to resolve the most challenging supraordinal nodes of the fungal tree of life. Based on the comparison of 39 fungal genomes, 71 potentially single-copy orthologous genes were selected, and a total of 243 universal primer pairs were designed and tested on six exemplar species representing Ascomycota, Basidiomycota and early-diverging fungi. The successful amplification and sequencing of 19 new gene regions adds a total of ca. 13,000 bp per taxon, in addition to the commonly sequenced nucLSU, nucSSU, mitSSU, MCM7, RPB1 and RPB2. The final datasets used to evaluate the performance of these new genes include 37 non-lichenized fungi, for which genomic sequence data are available, and up to 24 lichen-forming members of the Arthoniomycetes, Dothideomycetes, Eurotiomycetes, Lecanoromycetes, and Lichinomycetes, for which the sequences were obtained mostly from cultures of the mycobionts using single-gene Sanger sequencing, but also through next generation genome sequencing. Maximum likelihood analyses were completed on each of the 19 novel and 8 commonly used gene regions independently and on various multi-locus combinations on the same set of taxa when possible. Phylogenetic efficiency (i.e., the level of resolution and internode robustness) and phylogenetic informativeness (sensu Townsend) among all loci was compared. The performance of novel versus older genes used to infer phylogenetic relationships among fungi is also compared.

Contributed Oral Session 2
Day: Monday, 16 July
Location:
Time: 0945

: A systematic revision of Clavariaceae (Agaricales) from the Pacific Northwest

Joshua M Birkebak* and Brandon Matheny. Department of Ecology and Evolutionary Biology, University of Tennessee, 332 Hesler Biology Building Knoxville, TN 37996-1610.

The diversity of Clavariaceae has been underestimated in the Pacific Northwest of North America and thorough taxonomic revisions are much needed. Molecular methods of species recognition have not been extensively used in the Clavariaceae and are found to elucidate species level diversity previously unidentified. Several species complexes exhibiting high morphological and sequence diversity have been observed. Morphological differentiation of some molecularly distinctive species is not yet possible, and cryptic species may be present. A monographic revision of the Clavariaceae in the Pacific Northwestern North America with taxonomic keys, illustrations and descriptions is being compiled and will be published pending further investigation. At present, twenty-eight species in the family have been identified based on herbarium and recent field collections. Ten of these are tentatively considered undescribed. Overall, the Pacific Northwest is represented by two species of Camarophyllopsis (one new), eight species of Clavaria (one new), one species of Clavicorona, four species of Clavulinopsis (two new), four species of Mucronella, and nine species of Ramariopsis (six new).

Contributed Oral Session 10
Day: Tuesday, 17 July
Location:
Time: 1615

: Rust fungi of Himalayan Moist Temperate forests of Pakistan, their diversity and distribution: An overview

Najam-ul-Sahar Afshan*, Abdul Nasir Khalid, and A R Niazi. Centre for Undergraduate Studies, University of the Punjab, Quaid-e-Azam Campus, Lahore, 54950, Pakistan.

Himalayan Moist Temperate forests of Pakistan have coniferous species, mixed evergreen and deciduous patches and broad leaved forests. These forests of Pakistan merge downward with the tropical thorn forests and upwards with the alpine meadows. The precipitation may exceed 600 mm, thus making of these montane forests a bio-geographical crossroad between submontane and alpine meadow vegetation. Being rich in plant diversity, these forests harbor a large number of rust fungi that are obligate parasites of plants. Although the fungal flora of Pakistan has been explored by several workers in the past, the important group of rust fungi has largely been neglected resulting in a paucity of literature and very fragmentary knowledge of these fungi, particularly in Himalayan Moist Temperate forests of Pakistan. This study was undertaken to explore and assess the diversity and distribution of rust fungi along with their respective host plants in this floristically rich area. This preliminary study of the hosts and their rusts includes approximately 169 species and 19 genera of rust fungi on approximately 260 species of host plants. These rust fungi include one (01) species each of genus Cronartium, Hyalopsora, Miyagia, Monosporidium, Pucciniastrum, Pucciniostele, Thekopsora and Uredinopsis respectively; two (02) species each of Gymnosporangium, Peridermium, Pucciniastrum and Uredo; three (03) and four (04) species of Caeoma and Coleosporium respectively; seven (07) of Melampsora; nine (09) of Aecidium; eleven (11) of Phragmidium; twenty four (24) species of Uromyces and ninety five (95) species of largest genus of rust fungi, Puccinia. This study enlists rust fungi of Himalayan Moist Temperate forests of Pakistan and presents their species richness and geographical distribution. This work will help to prepare a checklist of rust fungi of this area and will ultimately lead to the documentation and preparation of monograph of the rust fungi of Pakistan.

Contributed Oral Session 2
Day: Monday, 16 July
Location:
Time: 0930

: A phylogenetic evaluation of the tribe Leucopaxilleae: polyphyly, the LPD grade, and novel taxa from the Southern Appalachians

Marisol Sanchez-Garcia* and P Brandon Matheny. Department of Ecology and Evolutionary Biology, University of Tennessee, Knoxville, TN, 37996-1610, USA.

The tribe Leucopaxilleae Singer comprises eight genera that have been placed at various taxonomic ranks. Phylogenetic analysis of a four-gene region supermatrix (nuclear rRNA regions, rpb2) shows the tribe to be highly polyphyletic, with genera distributed in the Tricholomatoid, Marasmioid, Hygrophoroid and Pluteoid clades. Leucopaxillus, Porpoloma and Dennisiomyces, occur in the Tricholomatoid clade. Dennisiomyces, primarily a neotropical genus, has not been previously considered part of the Leucopaxilleae. We present preliminary data that suggest Dennisiomyces is closely related to Leucopaxillus and Porpoloma forming the 'LPD' grade from which the ectomycorrhizal (ECM) genus Tricholoma appears to be derived. These results also suggest that Leucopaxillus and Porpoloma are polyphyletic. Additionally, three unknown species that form a clade nested in the 'LPD' grade have been collected recently from the Southern Appalachians of the southeast United States. They are distinguished from other species in the LPD grade by possession of distinct cheilocystidia and pleurocystidia, presence of clamp connections and inamyloid, smooth spores. At present, we are unable to ascribe any known genus to this clade.

Contributed Oral Session 6
Day: Tuesday, 17 July
Location:
Time: 0900

: Potential of the trichomycete fungus Harpella melusinsae to inhabit the midgut of mosquito hosts.

Charles E Beard1* and Abdullah Inci2. 1Clemson University, School of Agricultural, Forest, and Environmental Sciences, 114 Long Hall, Clemson, SC, 29634, USA, 2Ericyes University, Faculty of Veterinary Medicine, Parasitology Department, Kayseri, Turkey.

Trichomycete fungi are symbiotic inhabitants of the guts of aquatic Diptera. The midguts and hindguts of Chironomidae (midges) and Simuliidae (black flies) have trichomycetes, whereas only the hindguts of Culicidae (mosquitoes) have trichomycetes. We asked why no trichomycetes use the midguts of mosquitoes even though the midguts are similar among the three families. We hypothesized four broadly defined limitations that prevent trichomycetes from occupying mosquito midguts: behavioral, structural, physiological, or host habitat. The first three limitations were investigated by exposing mosquitoes to Harpella melusiane, which normally grows only in black fly larval midguts. Larval black flies from the field sites were allowed to feed in glass jars and shed frass that contained trichomycete spores. Mosquito larvae were then exposed to the water containing shed Harpella melusinae trichospores. They were allowed to feed on the spores, and then assayed for colonization. We found that Harpella melusinae grew in the mosquito midguts (prevalence up to 66%). These results suggest that mosquito behavior, gut structure, or physiological limitations do not explain the lack of trichomycete fungi in the midguts of field-collected mosquitoes. We suggest that host-habitat limitations are probably important in limiting trichomycete colonization of mosquito midguts.

Contributed Oral Session 3
Day: Monday, 16 July
Location:
Time: 0830

: Nuclear condition of Aspergillus flavus by heterokaryosis and ploidy analysis

Farhana Runa1*, Ignazio Carbone1, Deepak Bhatnager2, and Gary A Payne1. 1Department of Plant pathology, North Carolina State University, Raleigh, NC, 2Southern Regional Research Center, USDA , New Orleans, LA.

Conidia of Aspergillus flavus are multinucleate but it is unknown whether they are predominantly homo- or heterokaryotic. A heterokaryon could serve as a source of genetic diversity within strains for pathogenicity and aflatoxin production if homokaryotic as well as heterokaryotic nuclei are packaged selectively into conidia during conidiation. To study nuclear condition in A. flavus we transformed strain AFC (arg7, pyrG) with either plasmid HH2B-ECFP to create AFC-ECFP (pyr), a strain expressing nuclei with cyan fluoresce, or with plasmid HH2A-EYFP to create AFC-EYFP (arg), a strain expressing yellow fluorescing nuclei. Protoplasts from the two strains were subjected to polyethylene glycol mediated cell fusion. Putative fusants were selected for their ability to grow on minimal medium, which should not allow the growth of neither nutritional mutant AFC-ECFP (pyr) nor AFC-EYFP (arg). The putative fusants produced mycelia with some nuclei expressing ECFP and other nuclei expressing EYFP. We also observed that conidia from these fusants have three types of nuclei: only EYFP expressing, only ECFP expressing, or both EYFP and ECFP expressing. Ploidy analysis by flow cytometry indicated that the putative 11-2 fusant is heterokaryotic. Moreover, flow cytometry and merged fluorescence microscopy image data showed that the majority of conidia within 11-2 are homokaryotic, expressing either EYFP or ECFP in subsequent generations; however, a very small percentage of conidia maintain nuclei expressing both. Conidia having nuclei with merged fluorescence have been separated and sorted by FACS. We will determine whether the merged fluorescence microscopy images are indicative of nuclei expressing EYFP+ECFP, maintained predominantly as heterokaryons or as diploids. Confocal microscopy will be applied to track the migration of EYFP+ECFP nuclei during germination and conidiation. This work will allow us to understand the mechanism whereby heterokaryons or diploids are maintained in multinucleated conidia of A. flavus.

Contributed Oral Session 11
Day: Tuesday, 17 July
Location:
Time: 1715

: Comparative transcriptomics and the arbuscular mycorrhizal fungi

Nicolas Corradi* and Stefan Amyotte. University of Ottawa, Ottawa, Ontario, Canada.

Arbuscular mycorrhizal fungi (AMF) are notorious for benefiting most land plants by establishing a widespread association with their roots; the mycorrhizal symbiosis. Despite their tremendous importance for terrestrial plants worldwide, however, the content and nature of their genomes has long remained elusive. Here, we report the acquisition of large-scale sequence data from the transcriptome of two different strains of AMF in the genus Rhizophagus (previously known as Glomus) using Illumina sequencing. A total of 39,000 contigs from these strains were assembled, annotated and compared against sequence data (over 40,000 contigs) recently acquired by others from the model AMF Rhizophagus (Glomus) intraradices. Overall, these high-throughput approaches resulted in the first large-scale comparison of biochemical processes occurring among closely related AMF strains; providing first-hand and long-awaited information about the diversity, evolution, origin and plasticity of the AMF proteome. In particular, our study revealed a number of evolutionary novelties and molecular mechanisms that were previously unknown to occur in the genome of these ecologically critical organisms.

Contributed Oral Session 5
Day: Tuesday, 17 July
Location:
Time: 0900

: Molecular systematics of saprophytic and plant pathogenic isolates of the cosmopolitan fungus Thelonectria discophora (Nectriaceae, Hypocreales, Ascomycota)

Catalina Salgado-Salazar1*, Amy Y Rossman2, and Priscila Chaverri1. 12102 Plant Sciences Bld, University of Maryland, College Park, MD 20742, 2USDA-ARS, Systematic Mycology and Microbiology Laboratory, Beltsville, Maryland 20705.

Thelonectria discophora, previously placed in Neonectria, (anamorph "Cylindrocarpon" ianthothele var. majus) is a cosmopolitan species found on a wide range of living or recently dead woody plants, including gymnosperms and dicotyledonous plants and less frequently on palm trunks and herbaceous materials. A variety of this species, "Neonectria" discophora var. rubi, has been associated with a distinctive basal canker of cultivated Rubus sp. Molecular techniques have been useful to define species limits and have revealed that many widespread and cosmopolitan organisms can be formed by multiple genetically distinct cryptic species or sub-species that may be geographically or ecologically correlated. In order to test if T. discophora is a group of cryptic species with geographic or ecological restrictions, multi-gene phylogenetic analyses were conducted using Maximum Likelihood and Bayesian Inference approaches on a worldwide collection of isolates. Fifteen cryptic groups could be distinguished within T. discophora distributed in three major clades. These clades were supported by high bootstrap values and posterior probabilities. Some cryptic groups were composed by isolates coming from close geographic locations; however geographic clustering is not the rule as isolates from distant regions also grouped together. Interestingly, saprophytic isolates on plants, soil and pathogens were found to form separate groups seemingly isolated by ecology. Genetic divergence and fixation index values indicate that the cryptic groups are highly divergent, not forming a single panmictic population.

This abstract is not yet assigned to a session.

: Stealth strategies of a cereal killer

Barbara Valent. Department of Plant Pathology, Kansas State University, Manhattan KS 66506-5502 USA.

Magnaporthe oryzae includes host-adapted populations that collectively threaten rice, wheat and other cereal crops worldwide. Past studies have focused on understanding development and function of the pathogen's appressorium, the dome-shaped cell that builds up and focuses enormous pressures to force a tiny penetration peg through the tough outer plant surface. Now, new understanding of host tissue colonization after penetration has emerged from live cell imaging of the fungus expressing fluorescently-labelled components during invasion of optically-clear rice sheath cells. The blast fungus colonizes its host using specialized intracellular hyphae that successively invade living plant cells. Filamentous hyphae that enter each rice cell invaginate the host plasma membrane, and then differentiate into enlarged, bulbous invasive hyphae that are cloaked in a tight coat of plant membrane. Fungal cells directly involved in the morphological switch are associated with a highly-localized biotrophic interfacial complex (BIC) that appears to function as the staging center for translocation of effectors, pathogen proteins that hijack host processes, into the cytoplasm of the invaded host cell. Indeed, some of the translocated effectors move ahead into neighboring rice cells before the fungus enters them, possibly preparing rice cells before invasion. The thickened invasive hypha again undergoes extreme constriction, forming a tiny penetration peg-like structure for crossing the plant cell wall into the next cell. The fungus searches for locations to cross the wall, suggesting that it recognizes and manipulates the plant's plasmodesmata, the tiny communication channels between plant cells, for its cell-to-cell movement. Knowledge important for controlling a major disease threat to global food security comes back down to understanding fundamental mechanisms for hyphal polymorphism, for fungal protein secretion and targeting, and for sensing and adapting to harsh environments.

Symposium 3 - The mycologist's guide to the new International Code of Nomenclature for algae, fungi, and plants
Day: Monday, 16 July
Location:
Time: 1600

: A mycologist's guide to the Melbourne Code.

Lorelei L Norvell1* and Scott A Redhead2. 1Mycotaxon, PNW Mycology Service, Portland OR 97229-1309 USA, 2National Mycological Herbarium, Eastern Cereal & Oilseed Research Centre C.E.F., Agriculture & Agri-Food Canada, Ottawa, ON K1A 0C6 Canada.

The International Code of Nomenclature for algae, fungi, and plants (ICN) replaced the International Code of Botanical Nomenclature (ICBN) during the 2011 International Botanical Congress (IBC) in Melbourne. The new ICN permits diagnoses/descriptions in English (no longer requiring Latin) and electronic publication of fungal names (via PDFs in ISSN/ISBN numbered publications). With registration of fungal names required for valid publication in 2013, the IBC Nomenclature Committee for Fungi (NCF) is now moving to approve one or more nomenclatural registries. The ICN also excludes names of microsporidians and organisms treated in other Codes, refines typification procedures and spelling of sanctioned names, and recommends how to designate type cultures. The Melbourne Congress also referred mechanical typification issues and governance of fungal nomenclature to special committees and approved NCF recommendations on the conservation and rejection of notable fungal names. The new ICN does not permit multiple names for fungi with pleomorphic life cycles, which the ICBN (under Art. 59) previously allowed for non-lichenized ascomycetes and basidiomycetes. After 2013, alternative new names will be either illegitimate or not valid, although alternatives published through 2012 may be valid or legitimate. Although teleomorph- and anamorph-typified names will compete equally for priority, the ICN dictates delay in adopting anamorph-typified names with priority over commonly used teleomorph-typified names. To buffer the effects of this major change regarding alternative names, NCF-sanctioned subcommittees will compile and adjudicate lists of conserved/rejected names of non-lichenized fungi (not restricted to the Ascomycota and Basidiomycota) for approval by the IBC. The NCF and International Commission on the Taxonomy of Fungi (ICTF) - collaborating to issue joint guidelines, updates, and advice - recognize that mycological opinion over the Art. 59 change is still deeply divided and must be considered.

Contributed Oral Session 5
Day: Tuesday, 17 July
Location:
Time: 0845

: A new monotypic genus to accommodate Cosmospora vilior and related species

Cesar S Herrera1*, Amy Y Rossman2, Gary J Samuels2, and Priscila Chaverri1. 1University of Maryland, Department of Plant Sciences and Landscape Architecture, 2102 Plant Sciences Building, College Park, MD 20742, 2United States Department of Agriculture, Systematic Mycology and Microbiology Lab, Rm. 246, Bldg. 010A, BARC-West, 10300 Baltimore Ave., Beltsville, MD 20705.

Cosmospora sensu Rossman (Ascomycota, Hypocreales, Nectriaceae) has included nectroid fungi with small, reddish, KOH+, smooth, thin-walled, laterally collapsing when dry, non- or weakly stromatic perithecia. Recently, the group was found to be polyphyletic based on molecular data, and has been segregated into multiple genera. However, not all Cosmospora-like fungi have been treated systematically. Some of these species form a clade that includes C. vilior (= Nectria vilior) and many species often labeled as "Cosmospora sp." The objectives of this research were to designate an epitype for N. vilior, which has been misapplied based on examination of the type specimen, and determine its phylogenetic position within Cosmospora sensu lato and the Nectriaceae. A multilocus phylogeny was constructed based on six loci (ITS, LSU, RPB1, MCM7, TEF1, and TUB) to estimate the species tree. Results from the phylogenetic analyses indicated that C. vilior forms a monophyletic group with other Cosmopora-like fungi that have an Acremonium-like anamorph and that parasitize Eutypa and Eutypella (Diatrypaceae). The group is phylogenetically distinct from other previously segregated genera, and for that reason, a new genus is described to accommodate these species.

Contributed Oral Session 7
Day: Tuesday, 17 July
Location:
Time: 1430

: Chemical diversity across individual lichens

Anne Pringle1*, Shugeng Cao2, and Jon Clardy2. 1Organismic and Evolutionary Biology, Harvard University, Cambridge, MA, 02138, 2Harvard Medical School, Harvard University, Boston, MA, 02115.

Fungi often grow as modular networks, and fungi in the genus Xanthoparmelia form foliose lichens. Foliose lichens grow radially from a center. Edges are younger than the middle of a thallus, and to test whether modules of different ages are chemically equivalent, we took samples from the edges and centers of transects laid across lichens of different sizes and used HPLC to characterize chemical diversity. The modules of a single thallus are not equivalent. The same suite of compounds is found across an individual, but dramatic differences in amounts of the different compounds mean the diversity of chemicals is greatest at lichen edges, regardless of the size of the thallus. The centers of very large, old lichens are chemically depauperate, as compared to edges, but even in small individuals centers possess fewer amounts of compounds as compared to edges, suggesting the pattern is not driven by age, and is caused instead by the need for edges to maintain abundant chemical arsenals to colonize new habitats. These data provide a context for our previous work on bacterial diversity across the thalli of lichens, and are also contextualized by ongoing research on demographies and life histories of filamentous fungi.

Contributed Oral Session 6
Day: Tuesday, 17 July
Location:
Time: 1015

: Analysis of a putative sensory rhodopsin in the Chytridiomycota

Steven R Ahrendt* and Jason E Stajich. Department of Plant Pathology and Microbiology, University of California, Riverside, CA 92521.

Rhodopsin is a seven-transmembrane G-protein coupled receptor (GPCR) that responds to light through photoisomerization of a covalently bound 11-cis-retinal molecule. This action forces helical motion and subsequent signal transduction through activation of the coupled G-protein. Rhodopsin homologs have only been identified in metazoan lineages, and while Dikarya fungi are known to have opsins, phytochromes, and cryptochromes to sense light, this type of rhodopsin has only been found encoded in the genomes of the early diverging Chytridiomycota and Blastocladiomycota fungi. Previous and current work has shown that zoosporic fungi are phototaxic but the molecular mechanisms of light sensing in early diverging fungi has not been explored. Here we describe structural and functional analyses of rhodopsin proteins identified in two species of Chytridiomycota, the amphibian pathogen Batracochytrium dendrobatidis and the terrestrial saprotroph Spizellomyces punctatus. Comparative genomics analyses of rhodopsin and flagellum genes in the Chytridiomycota, Zygomycetes, and Dikarya show a correlation of flagella and rhodopsin presence across the fungi. Computational modeling of the B. dendrobatidis and S. punctatus proteins indicates that they both adopt the seven transmembrane helix formation typical of rhodopsin proteins. Additionally, structural features associated with rhodopsin proteins are observed as well. The B. dendrobatidis protein sequence is notably lacking the conserved lysine residue, however this residue is present in the S. punctatus sequence. Phototaxis assays were performed to determine in vivo function of the rhodopsin proteins. Ongoing work to characterize protein function through expression in Pichia pastoris may provide insight into the mechanism of retinal binding and photoisomerization in these chytrid rhodopsins.

Contributed Oral Session 6
Day: Tuesday, 17 July
Location:
Time: 1000

: Actin dynamics in Aspergillus nidulans

Laura A Quintanilla* and Brian D Shaw. Department of Plant Pathology and Microbiology, Program for the Biology of Filamentous Fungi, 2132 TAMU, Texas A&M University, College Station, TX 77843, USA..

Actin is a major cytoskeletal protein required for the polarized growth of filamentous fungi. To study actin dynamics in living cells, Aspergillus nidulans was transformed with the Lifeact reporter construct that has been used to document actin patch, cable, and ring dynamics in other systems. Lifeact is a 17 amino acid peptide derived from the Saccharomyces cerevisiae actin binding protein Abp140p. Localization patterns for strains expressing Lifeact from three different promoters displayed similar actin localization patterns and all strains that were tested grew and developed as wild-type. Lifeact labeled actin localized to a sub-apical collar of endocytic actin patches that was located approximately 2 µm from the apex. Cortical actin patches were also present along the periphery of the hyphae. Actin patches displayed anterograde and retrograde movement throughout the hypha. The presence of a dense, complex, and highly dynamic network of actin cables (SAW - Sub-apical Actin Web) located at varying distances from the apex was also observed. This network of actin cables was hypothesized to be associated with branch site or septation site selection. An alternative hypothesis was that the SAW acted as a diffusion barrier. Actin cables also spanned the length of growing hyphae and were associated with the Spitzenkörper. When a hyphal branch was newly formed, an actin cable network formed inside the parent hypha and then entered the newly formed branch. A complex of actin cables predicted septation sites and then condensed to form a double acto-myosin ring, which eventually depolymerized as septal wall material was deposited. Further experiments designed to test our hypotheses will be discussed.

Symposium 3 - The mycologist's guide to the new International Code of Nomenclature for algae, fungi, and plants
Day: Monday, 16 July
Location:
Time: 1720

: Nomenclatural databases as working tools for taxonomists: opportunities and challenges

Vincent Robert*, Joost Stalpers, and Pedro W Crous. CBS-KNAW, Fungal Biodiversity Center, Uppsalalaan 8, 3534CT Utrecht, The Netherlands.

During the 18th IBC held in Melbourne, important decisions were made to adapt the CODE to accommodate novel developments. One of them was to enforce registration in online repositories. This will guarantee easy access to newly published taxa and associated data. A number of issues remain to be addressed: 1. possibility of multiple repositories that could act as official registrars has major implications. Presently Mycobank (IMA) and Index Fungorum (CABI) are the only repositories, but more might emerge in the future; 2. coexistence of several repositories necessitates heavy synchronization to reach the desired aims. However, no collaboration rules are imposed; 3. decisions of the NCF are needed with regards to the data that have to be deposited (minimum and extended datasets); 4. future scientific developments will require regular adaptations of the system (e.g. environmental sampling, genomic data), with cost implications for repositories; 5. access to type information of taxa should be available and unequivocal to allow proper taxonomic revisions, especially in the light of one fungus one name; 6. registration of species data should be regulated, especially concerning types (epitype, neotype, lectotype) and barcodes; 7. updates from and relations with journals will have to be clearly established in order to prevent differences between deposits and publications; 8. taxonomic data are not always up to date and finding experienced curators willing to spend time on maintaining taxonomic databases is a challenge that should not be underestimated; 9. high availability of the system has to be ensured and no serious down-time is acceptable; 10. continuous and professional services will have cost implications with regard to both hard- and software. Such questions will have to be addressed and solved by the mycological community before January 1, 2013.

Contributed Oral Session 17
Day: Wednesday, 18 July
Location:
Time: 1045

: The effect of soil on the fungal root endophyte community of Zea mays

Jose Herrera* and Ravin Poudel. Department of Biology, 100 E. Normal, Truman State University, Kirksville, MO 63501.

Zea mays is, arguably, the most economically important grass domesticated by man. Yet, few studies have examined and fully characterized the fungal root endophyte community inhabiting the below-ground portion of the plant. Taxonomic molecular identification of fungal species based upon the internal transcribed spacer (ITS) region of rDNA obtained from roots reveal soil as a prominent factor determining the composition of microfungal communities within the roots of maize (Zea mays subsp. mays, previously fully sequenced variety B73) and its progenitor, teosinte (Zea mays subsp. parviglumis), grown in Missouri clay soil or Missouri clay soil mixed with desert soil from New Mexico. Fungal communities in maize and teosinte grown in mixed soils were dominated by different proportions of Hypocreales spp. and dark septate endophytes (Pleosporales spp., and Sordariales spp., primarily) while those grown only in Missouri clay soils were dominated by Glomerales spp. Moreover, fungal communities colonizing roots of Z. mays differed significantly from those colonizing roots of native (and co-habiting) fescue grass (Festuca arundinacea). Interestingly, Pleosporean sequences also include Paraphaeosphaeria spp., one of the dominant and common fungal species described in various grass species across North America. The presence of such cosmopolitan fungal root endophytes suggests that some fungal associations are general in nature and could function as evolutionary hinges that help plants develop adaptations to a variety of environmental challenges.

Contributed Oral Session 8
Day: Tuesday, 17 July
Location:
Time: 1330

: Linking fungal biology to restoration: Use of host-specific ectomycorrhizal fungi to restore whitebark pine, a 'threatened' species in western North America

Cathy L Cripps1*, Erin Lonergan1, and Cyndi M Smith2. 1Plant Sciences and Plant Pathology Department, Montana State University Bozeman MT 59717, USA, 2Parks Canada, Waterton Lakes National Park, P.0. 200, Waterton Park, AB TOK 2MO, Canada.

North America's only stone pine (Pinus albicaulis) is under consideration for 'endangered species' status in the USA and is assessed as 'endangered' in Canada awaiting legal listing. This five-needle pine forms extensive forests of magnificent old growth or gnarled krummholz at treeline in western North America. It has a unique ecology that requires bird dispersal of seeds; squirrels compete for seeds and grizzly bears raid squirrel caches in a complicated food web that includes ectomycorrhizal fungi (EMF). The invasive white pine blister rust and mountain pine beetle have reduced forests 12-70% in parts of their original range. Large restoration efforts are underway as summarized in the 2012 'Range-Wide Restoration Strategy for Whitebark Pine' that promotes plantings of rust-resistant nursery seedlings. However survival rates of previously planted seedlings are low. We are investigating the use of native EMF as a tool to enhance survival for various projects including "Working together to Restore Terrestrial Ecosystems" for Parks Canada. Our surveys of EMF with this pine have revealed limited EMF diversity and an important set of suilloids. Some of the suilloids also occur with stone pines in Europe and Asia; others appear limited to whitebark pine. Host-specific fungi could confer an advantage not available to competitive trees. However host-specific fungi are also at risk and could decline with the tree species making restoration more difficult. Screening of 26 native EMF showed Suillus species related to those used to inoculate cembrean pine in Europe to be effective colonizers in the nursery; Rhizopogon lagged in colonization. We are currently developing inoculation methods for effective colonization of this tree species. Large out-plantings of inoculated seedlings are being monitored in Waterton Lakes National Park to determine if survival is enhanced. Restoration recommendations specific to whitebark pine will be presented with preservation strategies for host-specific EMF.

Contributed Oral Session 5
Day: Tuesday, 17 July
Location:
Time: 0830

: Host conservatism or specialization? Patterns of fungal diversification are influenced by host specificity in Ophiognomonia (Gnomoniaceae, Diaporthales)

Donald M Walker1,2*, Lena Struwe1, Lisa A Castlebury1, Amy Y Rossman1, and James F White, Jr.1 1Department of Plant Biology and Pathology, Rutgers University, 59 Dudley Rd., New Brunswick, New Jersey, USA 08901, 2Systematic Mycology and Microbiology Laboratory, USDA Agricultural Research Service, 10300 Baltimore Blvd., RM. 224, BLDG. 010A, BARC-WEST, Beltsville, MD, USA 20705.

Species of Ophiognomonia are leaf- and stem-inhabiting perithecial fungi occurring on host plants in the Betulaceae, Fagaceae, Juglandaceae, Lauraceae, Malvaceae, Platanaceae, Rosaceae, Salicaceae, and Sapindaceae. A recent study identified 45 species of Ophiognomonia by integrating phylogenetic species recognition and the genealogical sorting index to indicate reciprocal monophyly of species lineages. In this study host plant patterns at the order, family, genus, and species level were linked with a multi-gene phylogeny of Ophiognomonia to better understand speciation events and host associations in this genus. Our objectives were to: 1) map host plant preference on a phylogeny of Ophiognomonia, 2) determine if patterns of fungal diversification are influenced by host plant association, and 3) determine if speciation events in Ophiognomonia are associated with host switching or host conservation at certain taxonomic host ranks. Host plant comparisons were made across 43 nodes in the phylogeny of Ophiognomonia using the program SEEVA (Spatial Evolutionary and Ecological Vicariance Analysis). Host/fungus correlations were interpreted using the divergence scaled index (D), with 0 equal to no divergence between sister clades, and 1 equal to the maximum possible divergence. Five deep nodes in the phylogeny of Ophiognomonia independently displayed D-values (0.62*-1.00*) at the host plant order, family, and genus level. Shallow nodes were often associated with the host plant genus and species. In summary, both host switching and conservation were observed and quantified for Ophiognomonia. Host specificity is suggested as a mechanism contributing to speciation in this genus. This method of analyzing host plant data provides a better understanding of evolutionary mechanisms influencing speciation events in the genus Ophiognomonia. A comprehensive picture of host/fungus evolution in Ophiognomonia provides a framework for evolutionary hypotheses about other genera in the Gnomoniaceae. This methodology could be useful to plant quarantine officials for making predictions about host specificity of plant pathogens.

Contributed Oral Session 4
Day: Tuesday, 17 July
Location:
Time: 1015

: Initial population abundance shaped the competition between two wood-degrading fungi

Zewei Song* and Jonathan Schilling. Department of Bioproducts and Biosystems Engineering, University of Minnesota, St. Paul, MN 55108.

Competition of wood-degrading fungi determines the structure of their community and the resulting ecological processes. During wood decomposition, brown and white rot fungi utilize different forms of carbon while competing for the same space. In this research, we studied the effect of initial population abundance on the competition outcome and residue properties of two wood-degrading fungi on four wood types. A microcosm system was used to allow brown and white rot fungi invade and compete on wood substrates (oak, birch, pine and spruce). The initial population size was controlled by the area of agar inoculated into the microcosm. A brown rot fungus (Gloeophyllum trabeum) was inoculated in larger amounts (646mm2) than a white rot fungus (Irpex lacteus, 77mm2), simulating a stronger brown rot population due to earlier colonization. Decay residues were harvested after 3 and 8 weeks. A previous experiment showed that when both fungi were inoculated at the same size, the white rot fungus always dominated the system. The wood residue after 3 weeks showed distinctive white rot properties, with low solubility (~20%) and high pH (~4). However, after 8 weeks, a proportion of wood residues showed distinctive brown rot properties, which were high solubility (~60%) and low pH (~2). Quantitative PCR also showed that at 3 weeks the white rot fungus was dominant in wood. But after 8 weeks, some of the residues showed much higher percentage of the brown rot fungus. It is possible that the brown rot fungus was unsuccessful in invading wood at an early stage of competition, but was able to survive in feeder strips and outcompeted the white rot fungus after 8 weeks. Our study indicated that initial population abundance has a profound effect on the competition outcome.

Contributed Oral Session 4
Day: Tuesday, 17 July
Location:
Time: 0830

: Fueling the future with fungal genomics

Igor Grigoriev. US DOE Joint Genome Institute, 2800 Mitchell Dr, Walnut Creek, CA 94598.

Until recently, genomic diversity of Fungi was biased towards ascomycetes of medical importance. To correct this bias the Fungal Genomics Program (jgi.doe.gov/fungi) of the US Department of Energy (DOE) Joint Genome Institute (JGI) has partnered with international scientific community to explore fungal diversity in several large scale genomics initiatives aligned with the 2010 Grand Challenges for Biological and Environmental Research: a long term vision. The first initiative, the Genomic Encyclopedia of Fungi, is focused on diversity among DOE relevant fungi in the areas of plant health, to explore the interactions of bioenergy crop species with symbionts and pathogens, and of biorefinery, to catalog industrially relevant genes, pathways, and hosts for bioenergy applications. The second initiative, the 1000 Fungal Genomes project, is aimed to explore fungal diversity across the Fungal Tree of Life in order to provide references for research on plant-microbe interactions and environmental metagenomics. The third initiative is to support functional studies of fungal systems of varying complexity: from new model organisms to metagenomes of complex communities. Open to all scientists around the world, these initiatives result in massive amounts of genomic information integrated with analytical tools and community-driven experiments. Several examples produced by these initiatives will illustrate how genomic data and different sampling strategies can improve our understanding of fungal diversity, interactions, and evolution.

Symposium 7 - Using cellular structure and biochemistry as indicators of fungal phylogeny
Day: Wednesday, 18 July
Location:
Time: 1740

: Regulation of conidiation in Aspergillus nidulans and Neurospora crassa

Brian D Shaw*, Da-Woon Chung, Srijana Upadhyay, Charles Greenwald, Sheng-Li Deng, Heather H Wilkinson, and Daniel J Ebbole. Program for the Biology of Filamentous Fungi, Department of Plant Pathology and Microbiology, 2132 TAMU, Texas A&M University, College Station TX 77843, USA.

Did conidiation arise more than once in ascomycete lineages or did extant condiation strategies diverge from an ancestral form? To begin to address these questions, transcriptional regulation of conidiation from the two best studied conidiating ascomycetes, Aspergillus nidulans and Neurospora crassa, was examined. We hypothesized that a conidiation regulatory pathway was present in the ancestral species, and became specialized in the extant species by gaining and/or losing specific regulators. N. crassa orthologs of seven regulatory genes in A. nidulans (fluG, flbC, flbD, abaA, wetA, medA, and stuA) were examined. Expression of the N. crassa orthologs complemented defective conidiation in the A. nidulans fluG, flbD, wetA, medA, and stuA mutants. We detected four patterns for the conidiation regulators: (i) Non-homologous genes with analogous roles in conidiation (brlA and fl), (ii) Orthologs with retained biochemical function that lack an analogous role in conidiation (fluG, flbD, and wetA), (iii) Orthologs with retained biochemical function with analogous roles in conidiation (medA and stuA), and (iv) Orthologs with biochemical function not conserved with analogous roles in conidiation (abaA). It is reasonable to expect that if the extant conidiation strategies for these two species evolved from the same ancestral pathway the sets of genes co-regulated with known targets of conidiation regulators would exhibit considerable overlap. However, transcriptional profiling of both species across a time course of conidiation using RNASeq revealed no overlap in co-regulaiton of these gene sets, thus providing no evidence for shared mechanisms below the regulators. Taken together, our data for the roles of conidiation regulator orthologs and the behavior of known conidiation associated genes that are likely targets of regulators across these distantly related species provides little support for a common pathway.

Contributed Oral Session 1
Day: Monday, 16 July
Location:
Time: 1015

: On the rocks: the first look at fungal biodiversity in marine biofilms of submerged artificial reefs

Amy Salamone* and Allison Walker. Department of Coastal Sciences, University of Southern Mississippi, 703 East Beach Dr., Ocean Springs, MS 39564.

This study was the first characterization of natural mixed-species fungal communities in biofilm on artificial reefs in the marine environment. Previous fungal biofilm studies have failed to observe the naturally occurring biodiversity in coastal areas, which are the most utilized zones of the marine environment. Fungi have evolved to form biofilms for protection, to communicate via quorum sensing, to gain access to nutrients, and to exchange genetic information. These communities fuel many primary consumers that support artificial reef habitats. Fungi, along with bacteria, archaea, protists, algae, and diatoms, quickly form a complex biofilm on immersed surfaces in seawater. This settlement and succession calls for further examination, given the major trophic implications therein, as properties of the biofilm can influence future establishment of other marine organisms. For this study, an optimized extraction technique for fungal DNA from biofilm was developed. Fungal biofilm communities were characterized by culturing, morphological identification, ITS T-RFLP analysis, and ITS gene sequencing. The role of seasonality and other abiotic factors in structuring these communities was also examined. Fungal biodiversity in these biofilms is high, spanning over 25 genera, including species of Ascomycetes, Basidiomycetes, Zygomycetes, and potential novel yeast species. This assessment of fungal community biodiversity in marine biofilm yields insight into artificial reef fungal succession and functionality in the North-Central Gulf of Mexico.

Contributed Oral Session 6
Day: Tuesday, 17 July
Location:
Time: 0945

: Investigation of sclerotial morphogenesis in the soil fungus Rhizoctonia solani

Elizabeth Thomas*, Nick Taylor, Pio Moises Figueroa-Contreras, Zhi Zhang, Kelly L Ivors, and Marc A Cubeta. Department of Plant Pathology, North Carolina State University, Raleigh, NC 27695.

The soil fungus Rhizoctonia solani (teleomorph=Thanatephorus cucumeris) is an important pathogen of cultivated and native species of plants. The fungus is a competitive saprobe that promotes the decomposition of organic matter and survives in soil in the absence of a host plant by forming sclerotia. Although there has been an accumulating body of knowledge generated on sclerotial forming fungi, little is known about the molecular mechanisms involved in the formation of sclerotia by R. solani. To examine the biochemical and physiological changes of the fungus during sclerotial morphogenesis, an in vitro assay was developed to induce sclerotia formation in a wild type strain of R. solani anastomosis group 3 (Rhs 1AP) and two protoplast-derived strains with a reduced nuclear genome (Rhs 1AP-115 and Rhs 1AP-123E) that had lost their ability to produce monilioid cells and sclerotia on nutrient media and potato tubers. The addition of xylose, arabinose, galactose, mannose, maltose, trehalose, raffinose, starch, cellulose, xylan oat-spelt, and casein to minimal medium stimulated radial mycelial growth of Rhs1AP but not Rhs 1AP-115 and Rhs 1AP-123E. The supplementation of minimal media with arabinose, mannitol, sorbitol, xylitol, glycerol, and myo-inositol induced formation of sclerotia in strain Rhs 1AP but not in strains Rhs 1AP-115 and Rhs 1AP-123E. Growth of the latter strains in the presence of cAMP or under nutrient deprived conditions also did not result in the formation of sclerotia. Results from the analysis of the transcriptome of strains Rhs 1AP (sclerotia plus) and Rhs 1AP-123E (sclerotia minus) will be discussed.

Contributed Oral Session 2
Day: Monday, 16 July
Location:
Time: 0830

: Are there New Zealand-only clades of rust fungi?

Mahajabeen Padamsee* and Eric McKenzie. Biosystematics Team, Landcare Research, Auckland, New Zealand.

Of the 250 species of rust fungi (Pucciniales) recorded in New Zealand, approximately 90 are believed to be endemic since they occur on native plants. Over the last 150 years, the number of non-native rusts has risen sharply from 33 to over a hundred. Although most of these rusts are recorded on non-native plants, several have been also recorded on native hosts. This begs the question, if host jumping has been observed in such a short time, what implication does this have for the origin of endemic rusts, i.e., are they really endemic? If not, then when and from where were the rusts introduced? Recent evidence suggests that many of the non-native rusts have been introduced from Australia by trans-Tasman airflows, which suggests that historically dispersal from Australia may have influenced the current distribution of rust fungi. A multi-gene phylogeny of New Zealand rust fungi was constructed and when combined with data from non-native rusts suggested the strong likelihood that there are New Zealand-only clades of rust fungi. Preliminary results indicate that several rust fungi may have evolved on closely related hosts in New Zealand.

Contributed Oral Session 6
Day: Tuesday, 17 July
Location:
Time: 0915

: Reaction partitioning by wood-degrading fungi - doing the two step without tripping

Jonathan S Schilling1*, Jun Ai1, Robert A Blanchette1, Shona M Duncan1, Timothy R Filley2, Joel Jurgens1, Justin Kaffenberger1, Gerald Presley1, and Urlike W Tschirner1. 1Department of Bioproducts and Biosystems Engineering, University of Minnesota, St. Paul, MN 55108, 2Department of Earth & Atmospheric Sciences, Purdue University, West Lafayette, IN.

Breakdown of lignocellulosic plant tissues such as wood is a key bottleneck in the global carbon cycle and is a step in optimizing bioconversion when the goal is a product such as a fuel. On the applied side, current practice involves chemical pretreatment and acid or enzymatic saccharification to prepare biomass for production steps such as fermentation. These early steps are typically expensive and frustrate many efforts to commercialize approaches on the 'biochemical platform' for bioconversion. Although consolidation of steps lowers costs, to date, chemical pretreatment is a required and costly stand-alone step. In nature, brown rot fungi are theorized to consolidate an oxidative chemical pretreatment with enzymatic saccharification of wood. As evidenced in its genome, Postia placenta produces a limited suite of cellulases, lacking genes for exoglucanases. Despite this limitation, it is still capable of rapidly degrading wood. According to the current paradigm, brown rot fungi such as P. placenta quickly reduce degree of polymerization of wood with little weight loss, using a reduction-oxidation pathway that yields highly reactive hydroxyl radicals through a chelator-mediated Fenton reaction (CMFR). While hydroxyl radicals would readily damage cellulases, the fungus is believed to conduct both reactions, enzymatic and oxidative, in the same wood sample. We are examining the spatial and temporal relationship of these two reaction systems, complemented by proof-of-concept biomimick experiments. The results have shown clear reaction partitioning at coarse scale in wood wafers, which will be presented. The outcomes from such fundamental inquiry in a novel application context are useful, given the traditional pest-oriented focus on these fungi and the targeted focus on wood strength loss. Our systems-based research should help better understand these fungi, not only for potential in application but as key players in the global carbon cycle.

Contributed Oral Session 2
Day: Monday, 16 July
Location:
Time: 1015

: Basidioascus: a new lineage of heat resistant and xerotolerant basidiomycetes

Hai Nguyen*, Nancy Nickerson, and Keith Seifert. Agriculture and Agri-Food Canada | 960 Carling Avenue | Ottawa, Ontario, Canada | K1A 0C6.

Basidioascus was originally described from soil in Australia by Matsushima, who considered the type species, B. undulatus, to be an ascomycete with some basidiomycete characters. We isolated this fungus commonly from soil in temperate areas or Canada and the United States, using methods designed to isolate heat resistant and xerotolerant fungi. Molecular phylogenetic analysis of rDNA loci reveals that Basidioascus is a member of the Basidiomycota, with a sister-group relationships with Wallemia, a xerotolerant basidiomycetous mould that contaminates food and house dust. Microscopy and molecular phylogenetic analysis of the internal transcribed spacer (ITS) barcode region support the recognition of B. undulatus and two undescribed species. Our interpretation of microscopic observations is that ontogenesis starts with the development of lateral or terminal clavate, probasidia on hyphae, which become obovate when mature. After self-fertilization, the basidium ruptures, producing 1-3 dark, thick-walled globose to subglobose basidispores near the surface of the agar, leaving behind a collapsed basidium.

Contributed Oral Session 7
Day: Tuesday, 17 July
Location:
Time: 1415

: Estimating selective pressure across the coding sequences of the model yeast Saccharomyces cerevisiae

Ning Li* and Jeffrey P Townsend. Department of Ecology and Evolutionary Biology and Program in Computational Biology and Bioinformatics, Yale University, New Haven, Connecticut 06520, USA.

The impact of natural selection on the genome has been well studied in diverse model species. However, genome-wide scans of polymorphism and divergence in synonymous and replacement sites within genes have yielded surprisingly few genes whose tallies reject a null hypothesis, especially in organisms with very high effective population sizes such as the model yeast Saccharomyces cerevisiae. The surprising dearth may arise because targets of selection are often likely to be restricted to a single functional domain or a few local, interacting sites within a protein. Unfortunately, most data sets lack sample size under previous methodologies to effectively detect selection at these levels, leading to a tyranny of the null hypothesis. Additionally, local variation in mutation rate of synonymous sites presents a challenge for fine-scale detection of a history of natural selection. To address these challenges, we have developed an approach that performs model-averaged clustering of intragenic polymorphism and divergence. Applying Poisson Random Field theory to the model-averaged polymorphism and divergence over all potential clusters, we graphically profile the model-averaged level of selection and its 95% confidence intervals for each site. Our genomic analysis of genes across the S. cerevisiae genome demonstrates that although much of the coding sequence is under purifying rather than adaptive natural selection, nearly 50% of coding sequence overall has been evolving under positive selection. Moreover, adaptively evolving sites are distributed across about 80% of the genes in the whole genome. The presence of extensive regions of purifying selection explains the finding that few genes are significant under traditional tests of polymorphism and divergence, even though many critical sites or regions within individual genes are playing important adaptive roles in an evolutionary history of adaptive natural selection.

Symposium 5 - The Fungal DNA Barcode and Beyond
Day: Wednesday, 18 July
Location:
Time: 1430

: Fungal barcoding database, innovative concepts

Vincent A Robert1* and Conrad Schoch2. 1CBS-KNAW, Uppsalalaan 8, 3584CT Utrecht, The Netherlands, 2NIH/NLM/NCBI, 45 Center Drive, MSC 6510, Bethesda, Maryland 20892-6510, USA.

In April 2011, a conference held in Amsterdam (The Netherlands) that included a large and representative panel of mycologists from across the globe contributed to the creation of a database of 2000 fungal species and almost 5000 strains belonging to most of the major lineages of the fungal kingdom. Six markers were retained (ITS, SSU, LSU, RPB1, RPB2 and MCM7) and used to compare their ability to provide reliable identifications and to be used as DNA barcodes. A paper was recently published (Schoch et al., 2012) suggesting using ITS as fungal reference barcode. The database used for this paper served as seed infrastructure for the Fungal Barcoding database and website (www.fungalbarcoding.org). Since the launch of the system, a number of new records and functionalities were added to the system. Also a number of additional websites have been created by mycologists that are using the same template (Aspergillus, Penicillium, Fusarium, Morchella, yeasts, etc). The Fungal Barcoding website serves as a central access point, together with Mycobank, to give access to pairwise sequences alignment tools against a number of reference repositories. Unknown sequences can be compared against the CBS database, BOLD, the Fungal Barcoding website and/or Genbank at the same time. Combined results are presented to the end-users in a user-friendly way. Polyphasic identifications can also be performed on several reference databases using morphological, physiological and/or molecular data. A new annotation system allows Internet visitor to suggest improvements of species and strains records to the curators of the databases. A number of interesting and unique features of the system will be unveiled during the meeting.

Symposium 2 - The science that underpins fungal conservation
Day: Monday, 16 July
Location:
Time: 1330

: Systematics and DNA barcoding of waxcap fungi

Bryn TM Dentinger1,2*, D Jean Lodge3, A Martyn Ainsworth1, Paul F Cannon1, and Gareth W Griffith2. 1Jodrell Laboratory, Royal Botanic Gardens, Kew, Richmond, Surrey TW9 3DS, UK, 2Institute of Biological, Environmental and Rural Sciences, Prifysgol Aberystwyth, Aberystwyth, Ceredigion Wales SY23 3DD, UK, 3Center for Forest Mycology Research, US Department of Agriculture, Forest Service, Northern Research Station, P.O. Box 1377, Luquillo, Puerto Rico 00773.

Fungi are crucial components of all ecosystems and their conservation requires strategies specifically tailored to them. However, a major stumbling block in fungal conservation is our lack of knowledge of their true diversity. In the UK, waxcap fungi (Hygrocybe s.l. and Cuphophyllus) have been the subject of long-term monitoring, they have excited the attention of non-specialists, their presence has led to designation of a few Special Sites of Scientific Interest in the UK, and they are included in the sites analyzed in the Important Fungus Areas survey. Moreover, all four national statutory conservation bodies have funded survey and monitoring of these fungi. However, over the last decades, application of molecular markers to species diagnosis in fungi has called into question the reliability of a strictly morphology-based approach. A key requirement for many fungi of conservation concern in the UK is the need for molecular diagnostic tools to assist in species definition (including recognition of cryptic taxa). For this project, we focused on generating DNA barcodes from waxcap fungi to assist in identification of cryptic species that may need to be considered for conservation management. New citizen-led survey work has contributed over 500 new collections across Great Britain. In addition, sequencing information was harvested from over 650 fungarium specimens representing most of the known species in the UK. Over 200 new ITS barcode sequences were analyzed in combination with all of the Hygrocybe ITS sequences available on GenBank plus unpublished reference sequences, revealing a number of cryptic species and species with identification problems in the UK and USA. A reevaluation of morphological characters correlated with distinct ITS clades will inform future rapid, field-based surveys and monitoring efforts. In addition, these sequences have helped in revising infrageneric names in Hygrophoraceae - the first full revision since Hesler and Smith (1963).

Contributed Oral Session 6
Day: Tuesday, 17 July
Location:
Time: 0930

: The importance of carotenoids for sexual development of fungi

Nina A Lehr1*, Zheng Wang1, Francesc Lopez-Giraldez1, Ning Li1, Frances Trail2, and Jeffrey Townsend1. 1Yale University, New Haven, CT, USA, 2Michigan State University, East Lansing, MI, USA.

Carotenoids are terpenes which belong to the most widespread group of compounds in nature and which are produced by plants, bacteria and fungi. Terpenes fulfill diverse functions such as protection against reactive oxygen species (ROS) and are beneficial for the nutrition of humans and animals. The orange pigmentation of the fungus Neurospora crassa derives from the accumulation of the xanthophyll neurosporaxanthine and other precursor carotenoids in response to asexual spore development upon exposure to light. Carotenoid biosynthesis in N. crassa has been studied intensively and it is known that blue light controls induction of carotenoid production in the mycelium, formation of protoperithecia as well as phototropism of perithecial beaks and perithecial polarity. Carotenoid biosynthesis involves five different enzymes, amongst which are three "albino" genes al-1, al-2 and al-3, encoding enzymes essential for carotenogenesis but not for growth. In its life cycle, Neurospora undergoes either an asexual or a sexual phase depending on environmental conditions. The asexual phase results in hyphal growth leading to the formation of macro- and microconidia while in the sexual phase complex, three-dimensional fruiting bodies, perithecia, are formed. So far there is no information on the linkage between carotenoid biosynthesis and the sexual life cycle. We have performed Illumina next generation sequencing of three Neurospora species over the time course of sexual development, followed by a comparative gene expression analysis with a pipeline we have generated to measure the levels of gene expression. We have quantified the amount of total carotenoids present in the mycelium at each individual time point. A screen of knockout mutants has revealed a linkage of carotenoid biosynthesis to the formation of perithecia.

Contributed Oral Session 2
Day: Monday, 16 July
Location:
Time: 0845

: The "Big Ditch" project: TransAtlantic mushroom disjunction tested using multiple taxonomic tools.

Ronald H Petersen* and Karen W Hughes. Ecology & Evolutionary Biology, University of Tennessee, Knoxville, Tn 37996-1100.

Written descriptions and illustrations by historical Euro-Scandinavian mycologists crossed the Atlantic to America, where they were matched to fungi of the New World. Doubts about the accuracy of name applications across the ocean have persisted, but molecular phylogenies now provide a new level of resolution. Our research targets fleshy fungi whose names originated in Europe, but which bear the same name and superficial form in eastern North America. Several examples can be identified in which DNA sequences reveal differences across the ocean, including saprophytic (Marasmius rotula; Baeospora myosura; Sparassis crispa) and ectomycorrhizal (Strobilomyces strobilaceus; Tricholoma populinum) fungi. In addition, it appears that genes for sexual compatibility and recognition are more highly conserved than the internally transcribed spacer (ITS), but morphological differences vary with the individual taxon. Implications of this research include the necessity for new names for heretofore cryptic taxa on both continents and resultant adjustment of biodiversity inventory reports.

Contributed Oral Session 9
Day: Tuesday, 17 July
Location:
Time: 1500

: Nucletmycea, Holozoa, Opisthokonta, and the problem with the "Entagled Bank"

Frederick W Spiegel. Department of Biological Sciences, University of Arkansas, Fayetteville, AR 72701, USA.

In the famous last paragraph of the Origin, Darwin states, "There is grandeur in this view of life...that...from so simple a beginning endless forms most beautiful and most wonderful have been, and are being, evolved." From this, in part, comes the hard to shake idea that the progenitors of lineages were both primitive and simple. By definition, the characters of the ancestor of a lineage are primitive. However, many aspects of an ancestor, even a deep ancestor, were very complex, not simple at all. The last common ancestor of living eukaryotes was a sexual organism that had to have had a flagellate stage in its life cycle. These characters had to have been true of the first opisthokont, and it was certainly more complex than many members of the extant opisthokonts. Rather than worry about whole organisms, perhaps it is better to compare sets of characters that we can hypothesize result from share ancestry. For instance, multicellularity has arisen numerous times in eukaryotes. In opisthokonts, multicellularity that includes cell migration has arisen at least three times, at least once in Holozoa (Metazoa), and at least twice in Nucletmycea (the sorocarpic Fonticula and the dikaryomycetes). Do the seeds for using cell migration in multicellular development come from characters of the last common ancestor of the extant opisthokonts? We do not yet know, but if we learn to "think outside the box" as comparative biologists, it will become easier to recognize those characters that need to be compared and to develop a set of model organisms and that allow hypotheses to be tested.

Symposium 7 - Using cellular structure and biochemistry as indicators of fungal phylogeny
Day: Wednesday, 18 July
Location:
Time: 1620

: What can nuclear division, spindle pole body, and septal pore characters reveal about fungal phylogeny?

David J McLaughlin*, TK Arun Kuman, and Rosanne Healy. Department of Plant Biology, University of Minnesota, St. Paul, MN 55108.

A major goal of the AFTOL2 project has been to integrate molecular and structural data for key taxa at critical branches in the fungal tree of life to better understand phylogenetic relationships and character evolution. Nuclear division, spindle pole body (SPB) and septal pore characters have supported molecular phylogenies, but structural data is often absent for major clades or for taxa at key branch points within the fungal tree of life. We will consider recent data related to a number of hypotheses addressed by AFTOL2 and how the results of structural studies bear on them. These hypotheses include multiple losses of the flagellum and the transition to SPB's in Fungi. In the Kickxellomycotina a ring-shaped SPB occurs in Coemansia reversa, which differs from SPB's reported for other subphyla of zygomycetous fungi. The hypothesis that Orbiliomycetes is the earliest diverging lineage of Pezizomycotina gains support from septal pore analyses within the fruiting body of an Orbilia sp. and from ascus characters. The link between Ascomycota and Basidiomycota based on similarities in septal pore structure remains unresolved despite new insights into septal pore structure in Neolecta irregularis (Taphrinomycotina) and Helicogloea spp. (Pucciniomycotina), while a new septal type has emerged in Agaricomycotina through serial section reconstruction of the septal pore apparatus of Wallemia sebi. The latter supports the early divergence of the Wallemiomycetes. These data are retrievable for character analysis from the AFTOL Structural and Biochemical Database, but the database is a work in progress with a need for greater community involvement to make it more complete. Major gaps remain in the analysis of nuclear division, SPB and septal pore characters with some phyla and subphyla unstudied and a major phylum greatly understudied.

Symposium 3 - The mycologist's guide to the new International Code of Nomenclature for algae, fungi, and plants
Day: Monday, 16 July
Location:
Time: 1620

: Ascomycete taxonomy and the outset of the one name era for fungi

Andrew M Minnis. Center for Forest Mycology Research, Northern Research Station, USDA-Forest Service, Madison, WI, 53726.

Multiple correct names, including one for each stage of non-lichen forming ascomycetes with pleomorphic life cycles, have been allowed under formalized Codes used to name fungi for a significant period of the history of mycology. Due to philosophical preferences of a large group of mycologists, the most important article that allowed for multiple names (Art. 59) was essentially deleted at the 2011 Melbourne International Botanical Congress. The replacement text (Art. 59) in the International Code of Nomenclature for algae, fungi, and plants allows only one correct name for non-lichen forming ascomycetes with pleomorphic life cycles. Although already enacted, these changes, which will formally and retroactively take effect in 2013 and have substantial impact on how mycologists and other users of fungal names will communicate about fungi, will be summarized. Pragmatic considerations will be noted and illustrated by examples of the impact on ascomycete taxonomy at the various ranks, especially in regards to priority, legitimacy, and validity of names as well as author citations and types. Some observations on what in general will be "the rise of the anamorphs" will be made. The means by which the mycological community can avoid disadvantageous changes to names during the transitional period to one name while maintaining a connection to important historical works will be outlined and a call for much needed participation and help in the process will be shared. The importance of studying the whole fungus and integrating the taxonomy of anamorphs and teleomorphs in future works will be emphasized.

Contributed Oral Session 3
Day: Monday, 16 July
Location:
Time: 1000

: Comparative transcriptomics reveals new Neurospora crassa genes important to perithecial development

Nina Lehr1, Zheng Wang1, Usha Sikhakholli2, Francesc Lopez-Giraldez1, Ning Li1, Frances Trail2, and Jeffrey P Townsend1*. 1Department of Ecology and Evolutionary Biology and Program in Computational Biology and Bioinformatics, Yale University, New Haven, Connecticut 06520, USA., 2Department of Plant Biology, Michigan State University, East Lansing, MI 48824.

In recent years, a plethora of genomic sequences have been released for fungal species, accompanied by functional predictions for genes based on protein sequence comparisons. However, identification of genes involved in particular processes has been extremely slow, and new methodologies for identifying genes involved in a particular process have not kept pace with the exponential increase in genome sequence availability. We have performed transcriptional profiling of five species of Neurospora and Fusarium during six stages of perithecium development. Because we maintained a strictly common medium across experiments, our transcriptomic data revealed solely evolved differences in the transcriptional basis of morphological changes. We estimated ancestral gene expression profiles and transcriptional shifts across this developmental process, facilitating identification of genes whose transcription had substantially and significantly shifted during the evolutionary process. We examined one hundred genes whose expression greatly increased in Neurospora crassa perithecial development compared to Neurospora tetrasperma, compared to Neurospora discreta, or compared to Fusarium spp. Phenotypes of knockouts of these genes included substantial changes in the timing and environmental sensitivity of perithecial development. These genes were not previously identified as candidates for function in perithecium development, illustrating the utility of this method for identification of genes associated with specific functional processes.

Symposium 6 - Fungi in a Changing World
Day: Wednesday, 18 July
Location:
Time: 1740

: Interactions among saprotrophic fungi and invertebrate grazers under climate change

Lynne Boddy. Cardiff School of Biosciences, Cardiff University, Cardiff CF10 3AS, UK.

Interspecific interactions between fungi drive changes in fungal community structure. The progress and final outcome of these interactions, i.e. deadlock or replacement, are affected by the abiotic environment, changes in temperature, water potential and CO2 regime sometimes completely reversing outcomes. Similarly, invertebrate grazers can exert selective pressures on fungal decomposer communities in soil by feeding selectively on certain fungi and reversing the outcomes of competitive interactions. For example, by feeding selectively on the cord-forming fungus Resinicium bicolor, isopods prevented the competitive exclusion of Hypholoma fasciculare and Phanerochaete velutina in soil and wood. Nematode populations also reversed the outcomes of competitive interactions by stimulating growth of less competitive fungi. Composition of fungal communities affects decomposition rate, but so also can fungal-fungal and fungal-invertebrate interactions themselves stimulate or inhibit decomposition and extracellular enzyme production. Climate change has the potential to alter the activity of, and interactions between, saprotrophic fungi and soil invertebrate grazers, with implications for decomposer community composition, decomposition, ecosystem regulation and carbon feedback.

Contributed Oral Session 10
Day: Tuesday, 17 July
Location:
Time: 1600

: Invasion biology of the butternut canker fungus Ophiognomonia clavigignenti-juglandacearum

Kirk Broders1*, Andre Boraks1, Laura Barbison2, John Brown2, and Greg Boland2. 1Department of Biological Sciences, University of New Hampshire, Durham, NH 03824, 2School of Environmental Sciences, University of Guelph, Guelph, ON N1G 2W1.

Butternut canker caused by the fungal pathogen Ophiognomonia clavigignenti-juglandacearum (Oc-j) remains the primary cause for range-wide mortality of butternut trees. The disease was first reported in Wisconsin in 1967, however the fungus may have been present for several years prior. Several questions still remain largely unanswered regarding the invasion by Oc-j. These questions include; how many times was the fungus introduced; where was it introduced; were more virulent strains introduced; and how was it able to spread so rapidly within the native butternut population. Therefore, our objective was to evaluate the invasion biology of this fungus including investigations into the ecology, epidemiology, and population structure of Oc-j in North America. To complete these objective flowers, developing seeds, and mature seeds of butternut, heartnut and black walnut were assessed as potential vectors of Oc-j; Sixteen isolates were evaluated for virulence on butternut, heartnut, and black walnut; and 100 isolates of Oc-j from across North America were used to analyze the population structure of the fungus. Bayesian analyses based on 16 SNP markers revealed that the Oc-j population in North American is composed of four genetically distinct clonal lineages, suggesting multiple introductions of Oc-j, through successive or simultaneous introductions of isolates having differentiated genetic backgrounds. Isolates of Oc-j recovered from heartnut and black walnut caused larger lesions on all three Juglans species compared to isolates originally recovered from butternut. Four of the five isolates, which caused the largest lesions on butternut belonged to a single genetic cluster. The pathogenicity data in combination with geographic and population structure data indicate this fungus was introduced into North America on multiple occasions, and a more virulent clonal lineage was recently introduced into Minnesota and Wisconsin and subsequently spread throughout the rest of the butternut canker population in North America.

Symposium 5 - The Fungal DNA Barcode and Beyond
Day: Wednesday, 18 July
Location:
Time: 1450

: The use of DNA barcode techniques to identify the constituents of teas and herbal dietary supplements

Damon P Little. Cullman Program for Molecular Systematics, The New York Botanical Garden, Bronx, NY 10458.

Dried fragmentary plant materials, such as those found in teas and herbal dietary supplements, are difficult to identify to species using only morphological characteristics. Fortunately, PCR amplifiable DNA can be extracted from most dried fragments and the resulting DNA barcodes can be used to reliably make identifications. For example, black cohosh (Actaea racemosa) herbal dietary supplements are commonly consumed to treat menopausal symptoms. Accidental misidentification and/or deliberate adulteration results in harvesting other, related, species that are then marketed as black cohosh. Some of these species are known to be toxic to humans. Two nucleotides in the plant barcode region consistently distinguish black cohosh from related species. Of 36 dietary supplements sequenced, 27 (75%) have a sequence that exactly matches black cohosh. The remaining 9 samples (25%) have a sequence identical to that of three Asian Actaea species (A. cimicifuga, A. dahurica, and A. simplex). Another example comes from commercial herbal tea products that are often composed of a number of plant species. Although barcode sequences could be generated for 60 of 71 (84%) herbal tea products examined, matching DNA identifications to listed ingredients was limited by incomplete databases, shared or nearly identical barcodes among some species, and a lack of standardized common names for all plant species. Analysis of the barcodes generated indicate that 25 (35%) of the herbal teas examined contained ingredients not mentioned on their labels.

Contributed Oral Session 2
Day: Monday, 16 July
Location:
Time: 0900

: Toward a monograph of Gyroporus

Naveed Davoodian. New York Botanical Garden, Institute of Systematic Botany, 2900 Southern Blvd, Bronx, NY 10458.

The genus Gyroporus is a group of poroid ectomycorrhizal mushrooms in the Boletales with representatives documented from every major continent except Antarctica. They are strongly implicated as symbionts with Pinaceae, Fagaceae, Myrtaceae, and Casuarinaceae and there is evidence for associations with Betulaceae, Salicaceae, Euphorbiaceae, and Fabaceae as well. Though the genus is fairly easily distinguished (by a yellow spore print, broadly circumferentially arranged stipe hyphae, and clamp connections), and some members have been known since before Fries, the group has never been monographed. Research is currently underway on a contribution toward a monograph of Gyroporus. A preliminary overview of the research is presented, with emphasis on problems in the study of this group. Several major nomenclatural problems exist within the group. Also, preliminary phylogenetic studies by other investigators have yielded equivocal results; this is further compounded by the problem of morphologically similar taxa exhibiting disjunct, multi-continental distributions. As such, some of the well-accepted species in the genus are suspected to actually be species complexes. A global approach with broader sampling has been adopted to address these issues, and recent fieldwork in Australia has generated important collections and likely new species.

Contributed Oral Session 2
Day: Monday, 16 July
Location:
Time: 1000

: Addressing the polyphyletic origin of Phanerochaete (Polyporales, Basidiomycota) based on a multi-gene dataset: how many lineages exist?

Dimitrios Floudas* and David S Hibbett. Biology Department, Clark University, Worcester, MA, 01610.

Phanerochaete Karst. is a saprotrophic basidiomycete genus of the Polyporales, which harbors white rot species that remove both lignin and carbohydrates from wood. One species in the genus, Phanerochaete chrysosporium, is a model system for the study of white rot biochemistry and its genome has been sequenced, revealing a wide range of genes related to lignocellulose degradation. Despite progress on deciphering the white rot mechanism of P. chrysosporium, we lack a deeper understanding of the phylogenetic relationships of the genus as a whole. The lack or simplicity of macro and micro-morphological characters of the basidiocarps has rendered the identification of Phanerochaete species and the delimitation of the genus perplexing. Molecular studies, based mainly on ribosomal genes, have shown that the genus as traditionally delimited is of polyphyletic origin nested in Polyporales, but only a few Phanerochaete species have been incorporated in multi-locus phylogenetic studies. The fragmented phylogenetic picture for Phanerochaete hinders understanding of the morphological, biochemical and ecological evolution in the genus. We are sampling here multiple loci across Phanerochaete and related genera, as a part of the PolyPEET project, in an attempt to provide the first higher-level phylogeny of the genus, to assess its relationships with other genera in the Polyporales, and reassess the phylogenetic distribution of morphological characters. The preliminary results indicate that Phanerochaete s.l. species are found in at least 6 lineages in the phlebioid clade, highlighting the necessity for nomenclatural rearrangements in the genus.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P8: Fungal root endophytes in barley, tomato, and eggplant

Jonathan P Carver* and Thomas J Volk. Department of Biology, University of Wisconsin, La Crosse, WI.

In recent years it has become apparent that fungal root endophytes are both ubiquitous in nature and can have significant effects on host plant fitness. In fact many fungal root endophytes have extraradical hyphae which may allow them to function in a manner similar to mycorrhizae. Many of these extraradical root endophytes (ERE) are also facultative associates and can be easily cultured in the lab. The ability of these endophytes to be easily cultured, along with their ability to improve plant growth, means that there is a great potential for using these fungi to improve agricultural productivity and sustainability. Despite this potential we still know very little about ERE ecology and diversity and even less about the EREs naturally present in agricultural systems. In this research project putative fungal endophytes were isolated from three crop plants grown in garden soil; barley (Hordeum vulgare), tomato (Solanum lycopersicum), and eggplant (Solanum melongena). From these plants a total of 119 fungal isolates were obtained. Microscopic traits and culture morphology were used to group isolates into operational taxonomic units (OTUs) likely equivalent to morphological species or strains. Twelve of the most abundant OTUs were separated for further study and identified by sequencing the ITS region. These endophytes were subsequently co-cultured with each of the three crop plants in axenic growth media under indoor lights for 30 days. Effect of endophyte inoculation on shoot and root biomass was assessed and endophytes were re-isolated in order to fulfill Koch's postulates. Additionally, Roots were stained with Trypan Blue and Sudan IV to confirm ERE habit of isolates and to compare percent colonization with effects on plant growth. The results of this study may be useful in improving agriculture by expanding our current knowledge of the specific extraradical root endophytes found in crop plants and their ecology.

Contributed Oral Session 17
Day: Wednesday, 18 July
Location:
Time: 1030

42: Phylogeographic analyses of epichloid endophytes in Hordelymus europaeus suggest repeated host jumps and interspecific hybridizations

Martina Oberhofer1* and Adrian Leuchtmann2. 1University of North Carolina-Greensboro, Department of Biology, 321 McIver Street, Greensboro, NC 27412, 2ETH Zurich, Institut für Integrative Biologie, Universitätstrasse 16, CH-8092 Zürich, Switzerland.

Epichloid fungal endophytes (Epichloë and Neotyphodium spp., Ascomycota) are systemic, perennial symbionts of diverse pooid grasses. These endophytes display an array of life history traits, which are intimately linked to host grass fitness, thus making them excellent models for studying speciation processes. Presumed jumps to new host grass species and subsequent somatic hybridizations are common among epichloid endophytes resulting in increased genetic variation. Selection may act on this variation and thus initiate speciation. We explored the epichloid endophyte diversity of Hordelymus europaeus, a rare European woodland grass species. We collected 30 grass individuals in each of 28 populations along a latitudinal transect over the entire distribution range and molecularly characterized isolated endophytes. Based on the sequences of tubB and tefA, six distinct epichloid taxa (interspecific hybrid or cryptic haploid species) emerged from multiple phylogenetic approaches. Four taxa were novel, two of these were interspecific hybrids and two were of nonhybrid origin. Previously known endophytes of H. europaeus are seed-borne and strictly asexual. However, one of the novel nonhybrid endophytes found in the glacial refugium of the Apennine peninsula reproduced sexually in cultured plants. This is the first case of a seed-borne, but sexually reproducing endophyte of this host. Based on a phylogeographic approach, we discuss the origin, and possible ancestral species, of the six epichloid taxa. Repeated host jumps and somatic hybridizations characterize the diversity of these endophytes. To date, no other grass species is known to host a larger diversity of endophytes than H. europaeus.

Symposium 6 - Fungi in a Changing World
Day: Wednesday, 18 July
Location:
Time: 1640

110: The role of fungi in mediating ecosystem responses to global change

Kathleen K Treseder* and Krista L McGuire. Dept of Ecology and Evolutionary Biology, University of California Irvine, Irvine CA 92697.

We address whether trait-based mechanisms determine which fungal species respond to global warming, and whether the species that proliferate are those that will target recalcitrant C compounds, thereby reduce long-term C sequestration. The phylogenetic breadth of each trait should determine the degree to which the two traits may overlap within species. We hypothesized that larger, more complex (i.e., more recalcitrant) organic C compounds will be decomposed by a narrower phylogenetic range of taxa than are more labile C compounds, because the enzymatic machinery required to break down more recalcitrant compounds may be more complex and should evolve less frequently. Conversely, we hypothesized that a relatively broad phylogenetic range of taxa would respond to warming, given that multiple physiological changes can confer adaptation. We tested these predictions in an Alaskan boreal ecosystem. We used DNA sequencing to identify fungal taxa that were positively or negatively affected by warming. In addition, we assessed the relative abundance of recalcitrant- and labile-C users in situ by using nucleotide analog labeling of DNA. We found that taxa that fungi that used lignocelluloses and tannin-proteins, which are relatively recalcitrant compounds, were more phylogenetically narrow than expected by random chance (net relatedness index or NRI = 0.6 and 1, respectively). Fungal taxa that occupied warmed plots were neutrally-related to one another, as were those occupying control plots. The two traits were linked within fungal taxa, with recalcitrant C users responding positively to warming (P = 0.026). Ultimately, warming may lead to decreases in long-term C sequestration in the boreal forest.

This abstract is not yet assigned to a session.

: Stuff in Duff: a metagenomic study of fungi in kauri forests

Mahajabeen Padamsee1*, Stanley E Bellgard2, Ian Dickie3, Scott Fraser4, Duckchul Park1, Bryan Stevenson4, and Bevan Weir1. 1Biosystematics Team, Landcare Research, Auckland, New Zealand, 2Biodiversity and Conservation Team, Landcare Research, Auckland, New Zealand, 3Ecosystem Processes, Landcare Research, Lincoln, New Zealand, 4Soils and Landscape Responses, Landcare Research, Hamilton, New Zealand.

Kauri (Agathis australis, Araucariaceae) is restricted in distribution to the northern tip of the North Island of New Zealand. Living to 2,500 years or more and having trunks to 7 m diam., kauri is a keystone species. Kauri was extensively logged and now less than 1% remains of undisturbed old-growth forests. Since the 1970s these trees have also been under threat from the exotic invasive, Phytophthora taxon Agathis (PTA) that causes kauri dieback. We are interested in understanding how the fungal community associated with the large amount of kauri duff (i.e., leaf litter) will be affected after infection of the tree by this exotic root pathogen. However, there has been no thorough study of the fungi associated with kauri and as a result we have no baseline data of the composition of a "healthy" fungal community in kauri leaf litter. We used pyrosequencing to study fungal biodiversity and assess changes in fungal community composition after invasion by PTA. The sampling scheme was optimized and several methods of analyzing the data were tested. A sequence database of known fungi was created with which to compare the metagenomic data.

Round Table - Toward a roadmap for fungal conservation research in North America: tools, data needs and collaborations
Day: Monday, 16 July
Location:
Time: 1600

: Integrated Digitized Biocollections (iDigBio): A national resource for digitization

Barbara M Thiers*, Deb Paul, Cathy Bester, and Jason Grabon. iDigBio, FLMNH, Dickinson Hall, Gainesville, FL 32611.

iDigBio, Integrated Digitized Biocollections, is the National Resource funded by the National Science Foundation for Advancing Digitization of Biological Collections (ADBC). Through iDigBio, data and images for millions of biological specimens are being curated, connected and made available in electronic format for the biological research community, government agencies, students, educators, and the general public. The mission of iDigBio is to develop a national infrastructure that supports the vision of ADBC by overseeing implementation of standards and best practices for digitization; building and deploying a customized cloud computing environment for collections; recruiting and training personnel, including underserved groups; engaging the research community, collections community, citizen scientists, and the public through education and outreach activities; and planning for long-term sustainability of the national digitization effort. In addition to the iDigBio central digitization HUB, there are partner institutions referred to as "Thematic Collection Networks" (TCNs) which consists of networks of institutions with a strategy for digitizing information that addresses a particular research theme. The current TCNs include InvertNet - An Integrative Platform for Research on Environmental Change, Species Discovery and Identification; Plants, Herbivores and Parasitoids: A Model System for the Study of Tri-Trophic Associations; and North American Lichens and Bryophytes: Sensitive Indicators of Environmental Quality and Change. Additional TCNs will be added yearly, with four new TCNs to be announced in April 2012. Through the iDigBio HUB cyberinfrastructure, compilation and the inter-linking of data from the TCNs and existing collaborative databases, there will be opportunities to address research questions and education interests regarding biodiversity, climate change, species invasions, natural disasters, and the spread of pests and diseases. New TCNs will be funded in succeeding years based on solicitations from NSF. The iDigBio HUB is based at the University of Florida (UF), in partnership with Florida State University (FSU).

This abstract is not yet assigned to a session.

: Molecular approaches profiling of soil fungal diversity and composition under different land and crop management systems

Tiehang Wu1* and Dan Chellemi2. 1Department of Biology, Georgia Southern University, Statesboro, GA 30460, 2USDA-ARS-USHRL, Fort Pierce, FL 34945.

Diversity and composition of soil and root colonizing fungi were examined in tomato (Lycopersicum esculentum) plots subjected to diverse land and crop management systems. Culture-dependent colony counting was used to identify communities of fungi colonizing roots, and length heterogeneity polymerase chain reaction (LH-PCR) analysis of internal transcribed spacer (ITS) profiles was used to characterize soil fungal communities. Diversity of soil fungi in bahiagrass (Paspalum notatum var. notatum 'Argentine') or undisturbed weed fallow plots was significantly lower when compared to diversity in organically managed plots. However, the diversity of root colonizing fungi was significantly lower in organic plots when compared to the bahiagrass or weed fallow plots. Multivariate analysis of root colonizing fungi and genetic ITS-1 profiles of soil fungi both indicated a higher degree of similarity among fungal communities in weed fallow and bahiagrass plots. Soil fungal communities in organically managed plots displayed a high degree of similarity to each other and were unique when compared to communities in other land management systems. A dominant 341-bp amplicon was identified in all soil fungal communities except those from organic plots. The amplicon was identified as Fusarium oxysporum by cloning and sequence analysis and confirmed by the LH-PCR amplicon size and sequences for a known Fusarium oxysporum isolate. Fusarium oxysporum dominated all communities of root colonizing fungi except those subjected to organic management practices. This study addresses the benefits of integrating molecular approaches to profile whole soil fungal community with a correlation to target the functional subset of root colonizing fungi.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P1: MSA Student Section

Mia R Maltz1*, Naupaka Zimmerman2, Kolea Zimmerman3, and Sydney Glassman4. 1University of California - Irvine, 321 Steinhaus Hall Irvine, CA 92697, 2Stanford University, 3Harvard University, Department of Organismic and Evolutionary Biology Cambridge, MA 02138, 4University of California - Berkeley,.

At the 2012 meeting this year in New Haven, the Mycological Society of America will be launching a Student Section. This student-run group within the MSA will provide opportunities for students to network with other students in their own fields and beyond. It will also be valuable for student members of MSA seeking connections with those performing cutting edge research in mycology and thus has the potential to inspire future collaborative research. The Student Section is open and inclusive; we welcome the participation of all students (and faculty!) in building this group. We look forward to your participation in our new mentorship program and hope you can also join us at the following events.Events:Sunday July 15th: Using fungi to get a jobJoin us as Harvard University mycology professor Anne Pringle leads our first MSA Student Section event. This professional development workshop will take place after the Foray and before the Welcome Reception on Sunday.Monday July 16th: First Annual Student Section MixerJoin us before the poster session as we gather in the poster area for a casual meet and mix with coffee, dessert, and other drinks provided. The mixer will go from 6:00-6:30 p.m.Mentorship Program:Mentorship programs are an excellent way to introduce students to other researchers with similar interests. In addition, mentors may assist in steering student research or by providing information about previous studies or future opportunities.For these reasons, all students who expressed interest on their registration form have been assigned a senior mycologist who will serve as their mentor for the duration of the New Haven meeting. Students will meet their senior mycologist mentor at the Student Section Mixer on Monday, July 16th.We appreciate your involvement in the commencement of the new MSA Student Section and look forward to seeing you all in New Haven!

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P2: The gut of arthropods: a source for isolation of new Trichomonascus species

Hector Urbina1*, Sung-Oui Suh2, and Meredith Blackwell1. 1Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA, 2Mycology and Botany Program, American Type Culture Collection (ATCC), Manassas, Virginia 20110, USA.

The gut of insects has been a source for the isolation of several unrelated groups of yeasts, including members of the Candida tanzawaensis and Candida kruisii clades and species related to Candida albicans (Lodderomyces clade). Recently, these clades have been significantly enlarged as a result of intensive exploration of gut yeast community. The genus Trichomonascus (Trichomonascaceae: Saccharomycotina) is currently composed of 25 yeast species found primarily on decayed and fresh fungal sporophores or in association with insects. During our survey of gut yeasts, we isolated 93 Trichomonascus strains from the gut of a number of beetle families, including Anthribidae, Erotylidae, Endomychidae, Mycetophagidae, Nitidulidae, Scarabaeidae, Staphylinidae, and Tenebrionidae collected from Panama and the southeastern USA. Our molecular and morphological characterization identified the insect-associated Trichomonascus taxa as Trichomonascus petasosporus and as many as 15 undescribed species. The new Trichomonascus species are related to Trichomonascus indianensis and Trichomonascus farinosus previously isolated from fungal sporophores, as well as Trichomonascus capitulata and Trichomonascus nivea isolated from plant tissues. The gut of fungus-feeding beetles has proven to be a rich source of a number of yeasts, including undescribed Trichomonascus species.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P3: Microfungi community associated with yeast agriculture ant Cyphomyrmex minutus

Mariely Medina Rivera and Matias J Cafaro*. Department of Biology, University of Puerto Rico, Mayaguez, PR 00681.

Fungus-growing ants belong to the tribe Attini. They cultivate a single fungus (Basidiomycota) colony as food. To protect their cultivar from specific parasites in the genus Escovopsis (Ascomycota: Hypocreales) the ants engage in another association with antibiotic-producing Actinobacteria. In addition, the ants have specific hygienic behaviors that include farming and grooming of the cultivar. The ants also rearrange and transport the refused organic material inside and out of the nest. Because the cultivar pathogen is more abundant at the refuse material of the nest of other Attini ants in comparison with the cultivar garden is very important for the health of the cultivar to maintain this behavior. Cyphomyrmex minutus is the only Lower Attini species reported from Puerto Rico that practices yeast agriculture. We have investigated the microfungi community from both, the yeast cultivar and the refuse material piles. Our main objective was to detect and identify the cultivar pathogen Escovopsis and other microfungi associated with C. minutus. We obtained 152 cultures from 26 nests collected in Cambalache Forest in Puerto Rico. We also extracted total DNA from refuse samples and amplified the ITS region and cloned the products. The microfungi community associated to C. minutus is diverse. Three major classes of fungi are represented in our isolates (Ascomycota, Basidiomycota and Zygomycota). Aspergillus, Bionectria, Fusarium, Microdochium, Penicillium and Pestalotiopsis were shared between the cultivar and refuse material among 48 different genera detected. So far C. minutus yeast cultivar does not show signs of infection with Escovopsis or any other pathogens. We hypothesize that Escovopsis may not be present in Puerto Rico or that yeast agriculture is an effective adaptation to prevent pathogen infection.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P4: Entangled and nearly deceived in a web of gut fungi: two intermingled, dimorphic Smittium species from Idaho

Justin W Gause* and Merlin M White. Boise State University, Dept. of Biological Sciences, Boise, ID 83725.

Since 2007 we have been prospecting for gut fungi near Boise, Idaho. The Boise River, which runs parallel to Boise State's campus, sporadically has been a convenient and favorable system for aquatic insect collection, despite that it runs through this metropolitan area. Collections have included midges (Chironomidae), which are hosts of several genera of Harpellales, including Smittium. In 2008, several collections of larval midges revealed an astonishing density of hindgut dwelling gut fungi. Upon morphometric analysis of vouchered slides, two species of Smittium, both with dimorphic trichospores, were found to be coexisting inhabitants in the midge population. In addition to the dimorphic trichospores, zygospores were also recovered. Zygospores are often the missing "piece of the puzzle" for taxonomic considerations, and we were fortunate enough to have them within a few dissections. However, these specimens were remarkably well developed and presented intermingled thalli, demanding careful scrutiny to tease apart the key morphological aspects belonging to each. We present our data on the two species of Smittium and compare them with records of putatively closely related Smittium species, with suggestions for future efforts with these and other host populations in pursuit of gut fungi worldwide, perhaps in an area near you?

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P5: Extending knowledge of Trichomycetes by fixing on preserved immature aquatic insects in Idaho

Nicole K Reynolds1*, Emma R Wilson1, Prasanna Kandel1, Matthew Laramie1, David Pilliod2, and Merlin M White1. 1Boise State University, Department of Biological Sciences, 1910 University Dr. Boise, ID 83725, 2U.S. Geological Survey, Snake River Field Station, 970 Lusk St. Boise, ID, 83706.

Trichomycetes (gut fungi) are obligate symbionts of various arthropods and have been found in marine, freshwater and terrestrial habitats on every continent except Antarctica. Minimally, gut fungi associate commensally with their immature aquatic hosts (including black flies, mayflies, stoneflies, isopods, and others) attaching to the chitinous lining of the mid- or hindgut, although relationships may shift depending on the situation. Both the geographic distribution and the biodiversity of gut fungi are vastly underestimated. Idaho is no exception, as it presents many opportunities for discovery in unique habitats, including the sagebrush steppe. Fourteen locations in remote forest streams of Idaho were surveyed for macroinvertebrates, which were immediately preserved in 95% ethanol upon collection. Inspection of fixed specimens revealed gut fungi were occasionally protruding beyond the anus of the host. Specimens were rehydrated, dissected, slide mounted and those vouchers of gut fungi were used for identification. Hosts included black flies, mayflies and a few stoneflies. Gut fungi are more typically dissected from living hosts. These specimens, fixed in the field, presented both challenges and rewards. Whereas some fungi naturally extend beyond the anus at maturity, such extension in black fly larvae is not well known. Interestingly, a new species of Genistellospora, which has unusually large tricho- and zygospores, and is being described from a separate, less remote sampling location, was also observed in many of the black fly samples. We suggest that examination of fixed specimens may present similar opportunities for discovery. Additionally, with these data collected from fixed specimens, it is clear that these and similar other habitats in Idaho provide a healthy repository and diversity of trichomycetes.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P6: Fungal symbionts suggest an alternative origin for the red turpentine beetle (Dendroctonus valens) invasion in China

Stephen Taerum1, Z Wilhelm de Beer2*, Tuan A Duong1, Min Lu3, Nancy Gilette4, Jianghua Sun3, and Michael J Wingfield1. 1Department of Genetics, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria 002, South Africa, 2Department of Microbiology and Plant Pathology, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria 002, South Africa, 3State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, P. R. China, 4Ecosystem Function and Health, PSW Research Station, 800 Buchanan Street, Albany, CA 94710.

Species invasions are among the greatest threats to biodiversity, ecosystem stability, and the global economy. Invasive species frequently co-invade with numerous symbionts, some of which are themselves problematic in their new environments. Interestingly, the symbionts associated with an invading species can be useful in discerning the possible origins of invasive pests. We used phylogenetic analyses to determine and compare the composition of fungal communities associated with a forest pest, the red turpentine beetle (Dendroctonus valens), in its native [eastern (ENA) and western (WNA) North America] and invaded (China) environments. In total, 28 different species of Ophiostomatalean fungi were isolated in the three regions: 14 spp. in China, 13 in ENA and 11 in WNA. Five fungal species were shared between RTB populations in ENA and WNA. Two species (O. ips and O. floccosum) occurred in both China and WNA, but both were present in very low numbers and have a global distribution. Three species were present in ENA and China, including O. abietinum, also with a global distribution, L. koreanum, thus far only reported from East Asia, and L. procerum. The first two species respectively constituted 1% and 6% of the ophiostomatalean isolates obtained in both regions, while L. procerum was the dominant fungus in both regions, constituting 50% of the isolates in ENA and 56% in China. This fungus is pathogenic to pine trees in China, but not in the USA, and was conspicuously absent from RTB in WNA. These findings suggest that the RTB may have been introduced into China from ENA, contrary to previous reports based on population studies of the beetle, suggesting that RTB was introduced into China from WNA.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P7: Effect of a dominant endophytic fungus, Phialocephala fortinii, on plant growth

Katrina Sandona1*, Terri Tobias1, Antonio Rosales1, Katherine Suding2, Robert Sinsabaugh3, and Andrea Porras-Alfaro1. 1Western Illinois University, 1 University Circle, Macomb IL. 61455, 2University of California Berkeley, 130 Mulford Hall 3144, Berkeley CA. 94720, 3University of New Mexico, 167A Castetter Hall, Albuquerque NM. 87131.

Dark septate endophytes have been detected in over 600 plant species worldwide. Phialocephala fortinii have been reported multiple times as a common dark septate endophyte in the alpine tundra, heathlands, and forests. The objective of this study was to determine the effect on seed germination and plant growth of Phialocephala fortinii isolates on different commercial plants. Fungi were isolated from roots that were collected at the Niwot Long Term Ecological Research Site in Colorado. Root isolates were identified using ITS rDNA and Phialocephala fortinii was selected for germination experiments. The fungus SS37 was plated in five culture jars containing malt extract agar with antibiotics. Five jars with no fungus were used as controls. After a week six surfaced sterilized seeds of Zea mays (corn) and Glycine max (soybean) where planted in each jar. Seeds were allowed to grow for one week. Plants were harvested from the jars; the roots and stems were measured. The fungus Phialocephala significantly stimulated the growth of corn seeds with an average number of roots of 8.3±0.3 (SE) with respect to 2.4±0.3 SE for the control (P<0.05). Soybeans with Phialocephala had an average number of roots of 8.6±1.4 vs. the control with an average of 1.9±0.3 add SE roots (P<0.05). Phialocephala significantly promoted the growth of two commercial plants. The use of endophytic fungi has potential to reduce the need for fertilizers. Additional studies are necessary to understand the nature of these symbiotic relationships between plants and endophytes.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P9: Effect of nitrogen pollution on root fungal endophyte communities from two co-dominant alpine tundra plants at Niwot, LTER

Sarah L Hicks1*, Emily Farrer2, Robert Sinsabaugh1, Andrea Porras-Alfaro1,3, and Katherine N Suding2. 1Biology Dept, University of New Mexico, MSC03 2020 1 University of New Mexico Albuquerque, NM 87105, 2Environmental Science, Policy & Management University of California at Berkeley Berkeley, CA 94720, 3Department of Biological Sciences Western Illinois University 1 University Circle Waggoner Hall 372 Macomb, IL 61455.

Due to pollution from Boulder and Denver, CO, Nitrogen (N) deposition is increasing at Niwot Ridge, LTER. Over the last decade, one of the co-dominant alpine tundra plants, Geum rossii, has experienced substantial dieback, while the other co-dominant, Deschampsia caespitosa, gains dominance. G. rossii dieback has been attributed to elevated soil N rather than inter-species competition. The mechanism by which N kills G. rossii is still unknown. We hypothesize elevated soil N causes shifts in endophytic fungal communities, and that these shifts are responsible for G. rossii death. We expect G. rossii communities to become more pathogenic and parasitic in response to N. G. rossii and D. caespitosa roots were sampled from N-addition and control plots. Samples were surface sterilized prior to DNA extraction and 454 titanium pyrosequencing of fungal ITS ribosomal DNA. Sequences were clustered into OTUs by 97% similarity in Qiime. OTU correlation with N was assessed with ANOVA and community composition correlation with N was assessed with RDA, using the nlme and vegan packages in R respectively. G. rossii endophyte communities only clustered by N treatment at species level. Community shifts occurred among closely related species, and most sensitive taxa responded to N positively, including taxa that belong to pathogenic groups. Deschampsia caespitosa communities clustered by N at all taxonomic levels, and community shifts occurred between distantly related species, so that whole fungal Classes responded to N. Most affected taxa responded negatively. These data indicate that endophyte communities of D. caespitosa and G. rossii respond substantially differently to N addition. Deschampsia caespitosa endophyte communities appear to change more fundamentally. These changes may be important to D. caespitosa's ability to adapt to changing nutrients. If so, these data may provide insight into the mechanisms by which abiotic soil factors alter above ground vegetation dynamics.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P10: In search for a function: Endophytic fungi in the alpine tundra and their effect on cultivated plants.

Terri L Tobias1*, Katrina P Sandona1, Antonio D Rosales1, Andrea Porras-Alfaro1, Robert L Sinsabaugh2, and Katharine N Suding3. 1Western Illinois University, 1 University Circle, Macomb IL. 61455, 2University of New Mexico, 167A Castetter Hall, Albuquerque NM. 87131, 3University of California Berkeley, 130 Mulford Hall 3144, Berkeley CA. 94720.

Plants in the alpine tundra are anatomically adapted to survive harsh environmental conditions. They also rely on endophytic and mycorrhizal fungi for acquisition of nutrients and potential protection against pathogens. Microbial interactions with plants are known to play an important role in maintaining the biodiversity in the alpine tundra. Besides their importance, very little is known about fungal endophytic abundance, taxonomic and functional relationships with dominant plants. The main objective of this research is to examine the mechanism of transmission, diversity and function of seed and root associated fungi in an alpine tundra site located at the Niwot Long Term Ecological Research Site in Colorado. Seed endophytes were compared with root and soil communities using sequencing and culturing techniques. Dominant fungi were also tested in germination experiments. Dominant plants in alpine tundra harbor a community of endophytic fungi dominated by Ascomycota. A total of 33 endophytes were isolated from surface sterilized seeds. Greenhouse experiments with a dominant root fungal isolate identified as Philocephala fortinii showed significant positive effects on both germination and plant growth in two commercially cultivated plants (Zea mays and Glycine max). The average number of roots for soybeans after eleven days was 8.6 roots per plant as compared to the control of only 1.9 roots per plant. Likewise corn showed similar results with an average of 6.8 roots compared to the controls with 4.8 roots after seven days. Modern agriculture faces the serious issue of feeding a growing population while preserving the environment. This research will not only help understand the role of these fungi in natural ecosystems but has the potential to provide new insights in plant health and management.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P11: Vertically transmitted endophytes from plants in the alpine tundra

Antonio Rosales1*, Terri Tobias1, Katherine Suding2, Robert Sinsabaugh3, and Andrea Porras-Alfaro1. 1Department of Biological Sciences, Western Illinois University, 2Department of Environmental Sciences, University of California, Berkeley, 3Department of Biology, University of New Mexico.

Alpine tundra plants are able to survive harsh conditions with the help of endophytes and mycorrhizal fungi. Endophytes are fungi or bacteria that live within plants without causing apparent damage. These symbiotic relationships between plants and endophytic fungi are known to have a direct impact on plant community structure, fitness, and diversity. Endophytes can colonize plants by either horizontal or vertical transmission and little is known about their abundance, taxonomy and function. The objective of this study was to isolate and quantify vertically transmitted endophytes from plant seeds that were collected at the Niwot LTER Site in Colorado. Six plant species (Geum rossii, Erigeron simplex, Artemisia scopulorum, Deschampsia cespitosa, Bistorta bistortoides, and Trisetum spicatum) were studied. Thirty seeds of each plant were surface sterilized and plated on malt extract agar with antibiotics. Fungi were sequenced using the ITS rDNA region. Thirty three endophytes were isolated from the seeds. Ten different morphotypes were present. E. simplex and D. cespitosa show the highest germination rates. Germination rates varied between 67% to 0% and colonization rates varied between 6% to 33%. B. bistortoides seeds did not germinate and showed the highest fungal colonization rates. Preliminary identification of some isolates using ITS rDNA shows that G. rossii and D. cespitosa seeds are colonized by a fungus closely related to Cladosporium uredinicola. Additional tests need to be conducted to determine the potential role that these seed endophytes may have in plant germination, seed establishment and the structure of plant communities in the alpine tundra.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P12: Formation of Aspergillus flavus sclerotia on corn grown under different drought stress conditions

Bruce W Horn1*, Ronald B Sorensen1, Marshall C Lamb1, Victor S Sobolev1, and Ignazio Carbone2. 1National Peanut Research Laboratory, USDA, ARS, Dawson, GA, 39842, 2Department of Plant Pathology, North Carolina State University, Raleigh, NC, 27606.

Aspergillus flavus is a major producer of carcinogenic aflatoxins worldwide in corn, peanuts, tree nuts, cottonseed, spices and other crops. Many countries have strict limits on the amount of aflatoxins permitted in human commodities and animal feed. Sclerotia produced by A. flavus serve several functions in the life cycle of the fungus: (1) resistance to adverse environmental conditions; (2) colonization of substrates through myceliogenic and sporogenic germination; and (3) sexual reproduction. Drought stress in corn results in an increase in aflatoxin contamination, but little is known about the effects of drought on sclerotium production. Corn was grown at Shellman, Georgia, in 2010 under different drought stress conditions consisting of: rainfall only (0% irrigation) and 33, 66 and 100% of the recommended irrigation amounts. Decreasing water availability showed a progressive reduction in grain yield (r = 0.96, p < 0.001) and an increase in aflatoxin concentration (r = –0.62, p < 0.05), but had no significant effect on percentage of ears showing Aspergillus section Flavi sporulation or percentage of ears with sclerotia. Of the 8706 ears examined, sclerotia (n = 6022) were recovered from 84 ears (1%). Sclerotia of A. flavus L (large sclerotial) strain were dominant (78 ears); less common were sclerotia of A. flavus S (small sclerotial) strain (8 ears), A. parasiticus (3 ears) and A. alliaceus (2 ears). The sexual stage was not detected in any sclerotia, including those of homothallic A. alliaceus, suggesting that sclerotia may require additional incubation after dispersal onto soil.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P13: Competition experiments to test the efficacy of Aspergillus flavus biocontrol strains

Geromy G Moore* and Kenneth C Ehrlich. Southern Regional Research Center, USDA, ARS, New Orleans, LA, USA.

Several non-aflatoxigenic strains of Aspergillus flavus have been in use for nearly a decade to inhibit colonization of important food commodities by toxigenic Aspergilli such as A. flavus and A. parasiticus. Annual application of these biocontrol strains is necessary because they do not appear to persist in the field. The reason(s) for this are unknown. One possible mechanism to explain the loss of an atoxigenic biocontrol competitor strain is that it could be recombining with the native toxigenic population and reacquiring toxigenic properties. A second possibility is that aflatoxin production increases the likelihood of survival for toxigenic strains in the soil, during overwintering, compared to that of an atoxigenic strain. To test the relative efficacy of these non-aflatoxigenic strains as bio-competitors, and to assess the reason for the loss of the field inoculum with time (post-treatment), we transformed six different biocontrol strains with a plasmid containing DNA for expressing an 'enhanced green fluorescent protein' (eGFP) tag. Competition experiments in petri plates have been set up to test the efficacy of the transformed biocontrol strains to colonize cotton seed. Tests included: GFP-transformed biocontrol strains vs non-transformed (homologous and heterologous) biocontrol strains, and GFP-transformed biocontrol strains vs toxigenic strains of A. flavus and A. parasiticus. These in vitro studies will test if the GFP tag diminishes the aggressiveness of the biocontrol strain, as well as the relative abilities of the individual strains to compete against toxigenic strains. If these tests show promise in using GFP-tagged strains to track the introduced fungi, subsequent studies will be done under conditions imitating the growing environment of the cotton plant in order to track the longevity and recombining potential of these biocontrol strains.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P14: Trichoderma viride volatile organic compound profile: The effect of individual components on Arabidopsis thaliana

Richard Hung. Plant Biology, Rutgers University, 59 Dudley Rd., New Brunswick, NJ 08901.

Several Trichoderma species are successful biocontrol agents. Their effects include plant growth promotion, stimulation of systemic defense responses, and protection from pathogens. Our research has shown that the volatile organic compounds (VOCs) of Trichoderma viride have plant growth promoting effects on Arabidopsis thaliana without physical contact. GC-MS analysis of the VOC profile of T. viride has resulted in 51 unique detectable compounds of which isobutyl alcohol, isopentyl alcohol, and 3-methylbutanal were the most abundant. Experiments have been performed to determine which components of the T. viride profile are responsible for the observed growth in A. thaliana. These compounds, in addition to mushroom alcohol (1-octen-3-ol) and geosmin, have been tested on model systems with A. thaliana. Exposure to 1-octen-3-ol has resulted in increased growth both above and below ground. The average root mass of control plants was 0.36g and the average mass of VOC exposed plants was 0.77g showing a 113% increase in plant mass. In addition there was a 60% increase in chlorophyll concentration (5.5mg/g control, 8.8mg/g test). Conductivity readings of the tested plants were the same as controls at 12% ± 2% indicating no cell wall instability. Trypan blue staining showed no necrotic lesions and hydrogen peroxide, a reactive oxygen species (ROS) that is precursor to necrotic lesions was not detected with 3,3'-diaminobenzidine (DAB) staining. Exposure to ethanol, another component of the VOC profile, resulted in reduced seedling formation (73%) as compared to control (92%). More seeds were unable to throw off their seed coat and complete seedling formation (15%) than control (2%) at the end of three days.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P15: Morphological, physiological and molecular characterization of different strains of Geomyces

Shiloh Lueschow1*, Lynnaun Johnson1, Robert McCleery2, Rod McClanahan3, and Andrea Porras-Alfaro1. 1Department of Biological Sciences, Western Illinois University, Macomb, IL, 2Wildlife Ecology and Conservation, University of Florida, Gainesville, FL, 3USDA Forest Service, Shawnee National Forest, Harrisburg, IL.

Geomyces destructans is a fungus responsible for the mycoses in bats known as White Nose Syndrome (WNS), which has already killed millions of bats and may be driving some species towards extinction. Different Geomyces species are commonly found in the natural environment such as soils and caves, but it is not clear why G. destructans shows greater pathogenicity. G. destructans is considered a psychrophilic fungus (optimal grow is at low temperatures), but little is known about the growth requirements of other closely related Geomyces strains and species. The main goal of this study was to characterize the growth rates of various Geomyces isolates from Illinois and compare them to the pathogen G. destructans. Fourteen Geomyces isolates including G. destructans were plated on sabouraud dextrose agar. Isolates were identified using ITS rDNA. The isolates were obtained from bats collected in Illinois before the arrival of WNS. Isolates were incubated at 25ºC and 6 ºC. Growth rates were measured weekly to determine differences between the various isolates and G. destructans. Geomyces destructans showed no growth after three weeks at 25º C. The other Geomyces isolates obtained from bats in Illinois showed growth after the first week at 25º C. In all cases except for G. destructans, the isolates grew faster at 25º C than at 6º C. Excluding G. destructans, the growth rate range for 25º C was 5 mm/week to 9 mm/week. The growth rate range for 6º C was 3mm/week to 5mm/week. Our study suggest that the psychrophilic (cold loving) nature of G. destructans may be an important factor influencing its pathogenecity.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P16: White-nose syndrome and Illinois bat hibernacula

Steven J Taylor1, Andrew N Miller1*, Anthony C Yannarell2, Joseph F Merritt1, Nohra Mateus-Pinilla1, Edward J Heske1, Vincent P Hustad3, H M Lin2, Joseph A Kath4, and Rod D McClanahan5. 1Illinois Natural History Survey, University of Illinois, 1816 South Oak St., Champaign, IL 61820, 2Dept. of Natural Resources and Environmental Science, University of Illinois, 1102 South Goodwin Ave., Urbana, IL 61801, 3Dept. of Plant Biology, University of Illinois, 505 South Goodwin Ave., Urbana, IL 61801, 4Illinois Department of Natural Resources, One Natural Resources Way, Springfield, IL 62702, 5US Forest Service, Shawnee National Forest - Hidden Springs RD, 602 North First St., Vienna, IL 62995.

We are monitoring the invasion of Geomyces destructans in bat hibernacula in Illinois. Our team includes three mammalogists, a mycologist, a microbial ecologist, a wildlife veterinary epidemiologist, and a cave biologist, and is assisted by several resource managers. We use molecular and culture-based approaches to evaluate dead and live-caught bats and cave and mine substrates for the presence of G. destructans, and describe the microbial and fungal communities of sampled animals and caves. Beginning in winter 2012, we will visit about eight hibernacula per year for three years, and sample active bats during summer. We are collecting swab and wing-punch samples from asymptomatic and symptomatic bats; soil, air, and various other substrate samples from hibernacula; and temperature, humidity, and light data to characterize cave environments. Our study will provide data on the occurrence and distribution of G. destructans in hibernacula on the leading edge of the spread of white-nose syndrome, and the microbial ecosystems in which it becomes established. Circumstances permitting (i.e., timing and extent of the invasion), we hope to better understand potential competing or synergistic interactions between G. destructans and microbial communities on bats and in caves that influence the establishment of G. destructans.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P17: Comparison of fungal communities associated with different bat species in Southern Illinois

Tabitha F Williams1*, Robert McCleery2, Rod McClanahan3, and Andrea Porras-Alfaro1. 1Biology Department, Western Illinois University, Macomb, IL 61455, 2Wildlife Ecology and Conservation PO Box 110430, University of Florida, Gainesville, FL 32611, 3United States Forest Service, Shawnee National Forest, Harrisburg, IL 62946.

Bats are indispensable to ecosystem stability; they play an important role in the control of insect populations and plant pollination. White Nose Syndrome (WNS), caused by the fungus Geomyces destructans, is a new disease infecting bats in the USA and Canada. WNS was first reported in 2006 in New York and has since then spread across the eastern and central United States where it had made its way to the Illinois borders. The major objective of this research was to identify, characterize and compare psychrophilic ("cold loving") fungal communities associated with different bat species in southern Illinois. Bats were trapped using a harp trapping technique following USGS National Wildlife Center protocols. Swabs were taken from the bat wings and were inoculated in situ on petri plates in MEA (Malt Extract Agar) with antibiotics. Samples were incubated at 6°C to select for psychrophilic fungi. We evaluated seven different bat species; approximately 20 to 30 colonies were obtained in each petri dish, with an average of four unique morphospecies per swabbed- area (2-3cm2). From the swab plates about 300 psychrotolerant pure cultures were obtained. DNA extractions, PCR and sequencing reactions were used to identify fungal cultures. Fungal communities were dominated by Ascomycota followed by Zygomycota and Basidiomycota. Common fungi in bat wings include Penicillium corylophilum, Cladosporium spp, Epicoccum, and Alternaria. Most abundant species were isolated multiple times from different bat species and caves. This research will increase our knowledge about all fungal communities associated with bats which are endangered and currently facing local extinctions.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P18: Stuff in Duff: a metagenomic study of fungi in kauri forests

Mahajabeen Padamsee1*, Stanley E Bellgard2, Ian Dickie3, Scott Fraser4, Duckchul Park1, Bryan Stevenson4, and Bevan Weir1. 1Biosystematics Team, Landcare Research, Auckland, New Zealand, 2Biodiversity and Conservation Team, Landcare Research, Auckland, New Zealand, 3Ecosystem Processes, Landcare Research, Lincoln, New Zealand, 4Soils and Landscape Responses, Landcare Research, Hamilton, New Zealand.

Kauri (Agathis australis, Araucariaceae) is restricted in distribution to the northern tip of the North Island of New Zealand. Living to 2,500 years or more and having trunks to 7 m diam., kauri is a keystone species. Kauri was extensively logged and now less than 1% remains of undisturbed old-growth forests. Since the 1970s these trees have also been under threat from the exotic invasive, Phytophthora taxon Agathis (PTA) that causes kauri dieback. We are interested in understanding how the fungal community associated with the large amount of kauri duff (i.e., leaf litter) will be affected after infection of the tree by this exotic root pathogen. However, there has been no thorough study of the fungi associated with kauri and as a result we have no baseline data of the composition of a "healthy" fungal community in kauri leaf litter. We used pyrosequencing to study fungal biodiversity and assess changes in fungal community composition after invasion by PTA. The sampling scheme was optimized and several methods of analyzing the data were tested. A sequence database of known fungi was created with which to compare the metagenomic data.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P19: Searching for clues of sexual reporoduction in the genomes of arbuscular mycorrhizal fungi

Rohan Riley1*, Alexander Idnurm2, Philippe Charron1, Yolande Dalpé3, and Nicolas Corradi1. 1Canadian Institute for Advanced Research, Department of Biology, University of Ottawa, 30 Marie Curie Priv. Ottawa ON Canada K1N 6N5, 2School of Biological Sciences 5007 Rockhill Road University of Missouri-Kansas City Kansas City, MO 64110, USA, 3Eastern Cereal and Oilseed Research Center, 960 Carling Ave. Ottawa, Ontario K1A 0C6.

Arbuscular Mycorrhizal Fungi (AMF) represent an ecologically important and evolutionary intriguing group of land plant symbionts that have been long regarded as an ancient asexual lineage. However, the recent acquisition of large sequence datasets has revealed the presence of an alternative scenario for their supposed long evolutionary clonal history; including the presence of cryptic sexuality. Here we report the identification of many AMF genes that are commonly linked with the presence of sex in many fungi across the genome of different strains of Glomus. These include homologues of MATA_HMG proteins, which to our surprise were found be extremely diverse in both number and sequence variant. This elevated diversity is unmatched by any other known fungal species, and preliminary evidence suggests that some of these loci might be involved in partner recognition in these ecologically important organisms.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P20: Reproductive genetics and development in the fungus Myceliophthora heterothallica, a thermophilic model for the Chaetomiaceae

Miriam I Hutchinson1*, Amy J Powell2, Kylea J Parchert2, Joanna L Redfern1, Andrea M Martinez1, Randy M Berka3, Eric Ackerman2, Blake Simmons4,5, Igor V Grigoriev6, and Donald O Natvig1. 1Department of Biology, University of New Mexico, Albuquerque, NM 87131, 2Sandia National Laboratories, Albuquerque, NM 87123, 3Novozymes, Inc., Davis, CA 95618, 4Sandia National Laboratories, Livermore, CA 94551, 5Joint BioEnergy Institute, Emeryville, CA 94608, 6DOE Joint Genome Institute, Walnut Creek, CA 94598.

Members of the Chaetomiaceae are among the most reported fungi in studies of biomass degradation. They are of interest for their abilities to produce thermostable carbohydrate-active enzymes, which led to the sequencing of genomes from two thermophilic species, Myceliophthora thermophila and Thielavia terrestris. Until now, there has been no genetically tractable model either for this family, or more generally, for thermophilic fungi. We have characterized reproduction in the thermophile Myceliophthora heterothallica towards the goals of establishing this organism as a model for the group and developing it as an expression platform. M. heterothallica was reported to be heterothallic based on the fact that matings between two strains resulted in the production of fruiting bodies and ascospores. Prior to our work, however, heterothallism had not been confirmed with independent assortment of mating loci and autosomal genes. We speculate that this lack of confirmation of true heterothallism resulted from a failure to obtain ascospore germination. We found that ascospores are resistant to germination at temperatures below 47-50°C. This discovery allowed us to confirm heterothallism and analyze the segregation of markers in crosses. Sequences from opposite mating types show that mating regions are conserved relative to other Sordariales. Interestingly, different stages of development have different temperature optima: ascospore germination occurs at 47°C and above, ascocarp formation is optimal at 30°C, and growth is optimal at 45°C. We have successfully crossed M. heterothallica strains from Indiana, New Mexico and Germany, and we are expanding the number of known strains by surveying across latitudinal and elevation gradients. In addition, we are developing methods for transformation and gene replacement. Our goal is to develop M. heterothallica as a model organism to study fundamental aspects of thermophily and the biology of Chaetomiaceae.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P21: Cloning, characterization and population analysis of the mating-type genes from Leptographium procerum and L. profanum

Tuan A Duong1*, Z Wilhelm de Beer2, Brenda D Wingfield1, and Michael J Wingfield1. 1Department of Genetics, Forestry and Agricultural Biotechnology Institute, University of Pretoria, Pretoria 0002, South Africa., 2Department of Microbiology and Plant Pathology, Forestry and Agricultural Biotechnology Institute, University of Pretoria, Pretoria 0002, South Africa..

Leptographium procerum and its sibling species, L. profanum, are ascomycetes associated with root-infesting beetles of respectively pines and hardwood trees. Both species are native to North America, although L. procerum has been introduced into Europe, New Zealand, and South Africa. The latest introduction of L. procerum was into China in association with the red turpentine beetle (Dendroctonus valens), where the fungus apparently contributes to killing of millions of native Chinese pine trees. As for many other Leptographium species, sexual states have never been observed in L. procerum and L. profanum. The objectives of this study were to clone and characterize the mating type loci of these fungi, and to develop markers that can be used to characterize the mating-type of individual isolates. To achieve this, we amplified a partial sequence of MAT1-2-1 using degenerate primers targeting the high mobility group (HMG) box. A complete MAT1-2 idiomorph of L. procerum was subsequently obtained by screening a genomic library using the HMG sequence as a probe. Long range PCRs were used to amplify the complete MAT1-1 idiomorph of L. procerum and both the MAT1-1 and MAT1-2 idiomorphs of L. profanum. Characterization of the MAT idiomorphs suggested that the MAT genes are fully functional and that individuals of both these species are self-sterile in nature with a heterothallic mating system. Mating type markers were developed and tested on a population of L. procerum isolates from the USA, the assumed center of origin for this species. The results suggested that cryptic sex is occurring or has recently taken place within this population. The information regarding the MAT genes and mating type markers developed in this study will now be used in studies considering the population genetics and origin of these fungi.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P22: Description of two anamorphic yeasts in the Ustilaginales

Sebastian Albu*, Tomas A Rush, and M Catherine Aime. Louisiana State University Agricultural Center, Department of Plant Pathology and Crop Physiology, Baton Rouge, Louisiana, 70803.

Two basidiomycete yeasts belonging to the Ustilaginales were isolated in 2011 from the leaves of several fern species in Baton Rouge, Louisiana. Based on a combination of assimilation tests and phenotypic characterization, Farysizyma sp. nov. SA209 (Anthracoideaceae) and Pseudozyma sp. nov. SA575 (Ustilaginaceae) represent previously undescribed anamorphs in the Ustilaginomycetes. Colony morphologies are initially yeast-like, subsequently developing pseudohyphal tufts around the growing margin. Maximum likelihood analyses using four loci, the internal transcribed spacer (ITS) region, large subunit (LSU) and small subunit of the nuclear rDNA cistron, and translation elongation factor 1-alpha indicate that both isolates belong within Ustilaginales. The LSU and ITS regions of these isolates were compared to sequences of other available Farysizyma and Pseudozyma anamorphs and to related teleomorphs. Farysizyma sp. nov. SA209 is part of a Farysizyma/Farysia clade in Anthracoideaceae containing all other known species of Farysizyma. Pseudozyma sp. nov. is sister to Sporisorium hwangense within a larger clade of predominantly Sporisorium species that includes the type (S. sorghi). Comparison of our data with phenotypic descriptions and sequence data from all known species of Farysizyma and Pseudozyma indicates that neither isolate has been previously described in the anamorphic yeast state. However, without sequence information from all known members of Ustilaginales, we cannot rule out the possibility that either of these isolates may represent a previously described teleomorphic Farysia or Ustilago/Sporisorium species.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P23: The genetic structure of mating type locus B3 in Pleurotus eryngii

Jae San Ryu*, Kyung Hee Kim, Min Keun Kim, Su Myung Chae, Hee Jeong Je, and Won Du Song. Eco-friendliness Research Department, Gyeongsangnam-do Agricultural Research and Extension Services, Jinju 660-360, Republic of Korea.

King oyster mushroom Pleurotus eryngii is a heterothallic homobasidiomycete with a tetrapolar incompatibility system controlled by the two mating type loci, A and B. RAPD and BSA were performed to screen primers linked to the specific mating types. Out of 100 random primers, OPS-03 showed the specific band, OPS-032100 co-segregated with AxB3. The mating type B3 locus was identified based on a SCAR marker derived from OPS-032100 from a genomic library of the dikaryotic strain KNR2312. Sequence analysis suggested the genetic basis for B3 locus in P. eryngii was similar to those in model mushrooms. Interestingly, 4 putative pheromone receptor genes, instead of 3 which are typical, and 6 putative pheromone genes were predicted. Every pheromone receptor was packed with two pheromone precursors in three receptors while the extra one receptor, pepr3 does not have any cognate pheromone beside its sequences. B3 locus specific RNAs were isolated and reverse transcribed further into cDNA to characterize their sequence and role as signal factors. qRT-PCR showed transcription levels of the pheromone and the pheromone receptor in monokaryons were much higher than in a dikaryon. Such a study could be useful for developing self-fertilizing monokaryotic strains using RNAi (knock-down) and for deciphering comparative gene expression.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P24: Microbotryomycetes: Higher-level classification and description of a new genus

Merje Toome* and M Catherine Aime. Department of Plant Pathology and Crop Physiology, Louisiana State University Agricultural Center, Baton Rouge, Louisiana 70803, USA..

Microbotryomycetes, i.e., the "anther smuts" are a class of diverse Pucciniomycotina for which higher-level (e.g., order and family) classification has been largely unresolved. Recently, phylogenetic reevaluation of Microbotryomycetes was prompted by the isolation of a previously undescribed ballistosporic yeast from a neotropical forest in South America. The isolate, recovered from the surface of a fern leaf, forms butyrous cream-colored colonies with polar budding cells on solid media and rosette-like formations in liquid media. Based on phylogenetic, morphological and physiological analyses a new genus and species, Blackwellia eburnea nom. prov., are proposed. Phylogenetic analyses show that B. eburnea is most closely related to Kriegeria eriophori, a sedge parasite, the aquatic fungus Camptobasidium hydrophilum, and several recently described anamorphic yeasts that have been isolated from various plant or psychrophilic environments. Blackwellia eburnea makes an interesting addition to this clade since it represents the only species known thus far from the neotropics; all other species are either from temperate or cold environments. This indicates that additional studies of fungal biodiversity from poorly examined habitats like ice, water and tropical plant surfaces could greatly contribute to the species discovery and provide valuable information for improving the taxonomy of predominantly anamorphic clades in Microbotryomycetes. A higher-level classification is proposed to accommodate this and other species that until now have been incertae sedis within the Microbotryomycetes.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P25: Estimating genetic diversity of Cronartium ribicola populations in Oregon by RAD marker genotyping

Katarina Sweeney*, Michael Freitag, and Jeffrey Stone. Oregon State University, Department of Botany and Plant Pathology, 2082 Cordley Hall, Corvallis, OR 97331-2902.

The pathogen, Cronartium ribicola, was introduced to North America in 1910 and found in SW British Columbia and NW Washington by 1922. Evidence of evolutionary change in North American C. ribicola since its introduction is apparent and spread of two virulent races pathogenic to WPBR-resistant genotypes of Pinus monticola and P. lambertiana have been observed. The C. ribicola vcr1 race is virulent to P. lambertiana carrying a major gene (Cr1) that confers resistance to "wild-type" C. ribicola and was first found in California in 1978. The vcr2 race is virulent to P. monticola carrying Cr2 that also confers resistance to "wild-type" C. ribicola, and was found in Oregon in the early 1970s. Neither vcr1 or vcr2 races are virulent to the resistant genotypes of the other species. Few studies have been completed to estimate the genetic diversity of natural populations of C. ribicola in western North America, but accurate characterization of host phenotypes in resistance breeding programs requires more detailed understanding of the diversity of C. ribicola populations. Resistance screening is carried out with wild populations known to be predominantly vcr1, vcr2, or "wild-type" C. ribicola. In order to gain a more precise understanding of the molecular basis of natural variation in wild-type strains of C. ribicola in Oregon, we are employing restriction site associated DNA (RAD) marker genotyping of C. ribicola aeciospores. RAD sequencing generates short sequence reads adjacent to restriction enzyme sites that serve to identify restriction polymorphisms between samples or can be used to identify the presence of SNPs. We are using aeciospore samples from multiple locations in Oregon. RAD mapping will allow us to relate the variation seen in pine host reactions to infection by C. ribicola with informative SNPs in the pathogen.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P26: ITS, problematic to amplify a rust barcode

Alistair R McTaggart and M Catherine Aime*. Department of Plant Pathology and Crop Physiology Louisiana State University Agricultural Center, 302 Life Sciences Bldg. Baton Rouge, LA 70803.

DNA barcoding offers a rapid and accurate means to identify fungi and other organisms in comparison to traditional, morphological methods. Rusts (Pucciniales, Pucciniomycotina), the largest group of phytopathogenic fungi, are challenging for both morphological and molecular identification. They have a dynamic life cycle with five spore stages that infect different, often unrelated hosts. They are obligate parasites and cannot be maintained in pure culture, thus pure DNA is difficult, if not impossible to obtain. A molecular barcode that could differentiate between rust species and identify cryptic taxa is a necessity for diagnosticians and plant pathologists who rely mainly on morphology and host associations for species identification. One genus of rusts, Coleosporium, are taxonomically challenging and infect several agricultural and ornamental plants, such as pine and frangipani. Within northern America and Asia, C. asterum is believed to form a species complex. This study used Coleosporium as a model organism to investigate: 1. The systematic relationships of eight species of Coleosporium in northern America, including the C. asterum species complex. 2. The utility of the ITS region as a molecular barcode for rust fungi. 3. The potential of other loci, such as the LSU region, as rust barcodes.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P27: Population genetic structure of Phytophthora cinnamomi Rands associated with Phytophthora root rot of avocado within California and the potential introduction of a new clonal lineage

Deborah P Pagliaccia, Elinor P Pong, Brandon M McKee, and Greg W Douhan*. Department of Plant Pathology and Microbiology, University of California, Riverside, CA 92521.

Phytophthora root rot (PRR) of avocado (Persea americana Mill), caused by Phytophthora cinnamomi, is the most serious disease of avocado worldwide. Previous studies have determined that this pathogen exhibits a primarily clonal reproductive mode but no population level studies have been conducted in the avocado growing regions of California (Ca). Therefore, we used AFLP and mating-type to investigate pathogen diversity from 138 isolates collected in 2009-2010 from 15 groves from the Northern and Southern avocado growing regions. Additional isolates collected from avocado from 1970 to 2007 as well as isolates from other countries and hosts were also used for comparative purposes. Three distinct clades were found based on UPGMA analysis of 22 polymorphic loci. One clade contained only A1 isolates collected from various hosts and locations. Two A2 clades were found; one clade contained both newer and older collections from Northern and Southern Ca whereas the other clade only contained isolates collected in 2009-10. A total of 16 genotypes were found with only 1 to 4 genotypes identified from any one location. The results indicate significant population structure in the Ca avocado P. cinnamomi population, low genotypic diversity consistent with asexual reproduction, potential evidence for the movement of clonal genotypes between the two growing regions, and a potential introduction of a new clonal lineage into Southern CA. Current studies are underway to investigate the phenotypic variation (in vitro and in vivo) within our collection of P. cinnamomi isolates to determine if these new findings will affect our efforts towards breeding PRR tolerant rootstocks of avocado to control this important disease.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P28: Microsatellite markers reveal isolation by distance and recent population expansion of Waitea circinata var. circinata

Evans N Njambere1, Frank Wong2, and Ning Zhang1*. 1Dept. of Plant Biology and Pathology, Rutgers University, New Brunswick, NJ 08901, 2Bayer Environmental Science, Alexandria, VA.

Waitea circinata var. circinata (Wcc) is an emerging pathogen of turf grass in North America. It causes brown ring patch of turf grass in golf courses and amenity areas. The population dynamics and dispersal pathway of this pathogen in the US have not been well studied. In attempt to elucidate this, we isolated eight promising microsatellite markers from an enriched genomic library of Wcc. Seven of these markers were used to study the population diversity of eastern and western US populations of Wcc and the association with phenotypic characteristics. In general, the microsatellite markers were highly polymorphic, with an observed heterozygosity of > 0.5. The eastern population was differentiated from the western population but both populations showed the presence of immigrant individuals across boundaries. Analysis of population structure with geography revealed a significant clinal expansion, r2= 0.11, P = 0.01, of Wcc within a radius of about 150 km. However, it became more stochastic with increase in distance. No host specific clusters were observed based on UPGMA clustering, suggesting a lack of association between host and pathogen genotypes.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P29: A new species, and first record of Meliola weigeltii var. fraxinifoliae, on anacardiaceous hosts from the Brazilian Pantanal

William RO Soares, Eliane AS Armando, and José C Dianese*. Depto. de Fitopatologia, Universidade de Brasília, 70910900 Brasília, DF, BRAZIL.

A new Meliola species was found on leaves of Spondias lutea (Brazilian plum), a cultivated fruit crop also occurring in wild environments through the Neotropics. Our specimen was collected in the Pantanal Matogrossense (UB Mycol. Col. 21456) showing characteristic features related to: mycelial setae up to 267 × 5.5-9.5 µm, mostly erect, sometimes curved, with a broad apex when single containing 2-4 denticles up to 13.5 µm tall or 2-3 furcate with denticles up to 22 µm tall; ascomata 90-162 µm diam., superficial, glabrous, dark brown, globose to subglobose, spread on the superficial mycelium; ascospores 31-41 × 9.5-13 µm, oblong to elliptical with obtuse terminal cells, dark brown when mature, 4-septate, with constrictions at each septum. This specimen morphologically differs from all known Meliola species, strongly indicating that it belongs in a new taxon. This is also the forth record of a Meliola species on plants of the genus Spondias. Meliola weigeltii var. fraxinifoliae was found on Astronium fraxinifolium, which is the first record for the entire Pantanal Region, although it is already known from the Cerrado and from the Northeastern region of Brazil.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P30: An Evolutionary examination of ecological and physiological resistance within host-pathogen interactions: a Montiaceae and Microbotryum model

Jorge Mena-Ali, Erica Goldberger*, Elizabeth Heppenheimer, and Maggie Serpi. Department of Biology, Franklin & Marshall College, Lancaster PA 17603.

Historically, coevolution is considered the primary process to justify disease evolution between hosts and their pathogens. However recent research has highlight the relative importance of host shift events in the evolutionary divergence of pathogen species. The purpose of this project is to examine the evolutionary history of the association between smut fungi (Microbotryum spp.) that infect plants within the Montiaceae family. In this system, when fungal spores infect a susceptible host, a severe form of anther-smut disease develops, leading to complete sterilization of both male and female fertility in the plant. Through large-scale herbarium surveys we were able to analyze the prevalence and global distribution of the disease. To date, we have surveyed 11,864 sheets from 21 herbaria representing 206 species. Of these, 51 sheets showed disease giving a 0.43% disease rate. However, if we exclude those species without signs of disease, the disease rate increases to 9.9%. Additionally, we collected spore samples from infected specimens for molecular analysis. These samples were analyzed with fungal-specific markers (NADH, ITS). Initial phylogenetic analysis suggests a pattern of divergence among Microbotryum isolates that reflects host taxonomic classification. However, geographic distribution also seems to have contributed to isolation and speciation. These results will be used to analyze the evolutionary history of Microbotryum species associated with Montiaceae in the context of varying pathogenicity and host specificity.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P31: Fungi associated with Panicum spp. in New Jersey and Hawaii.

Emily Walsh* and Ning Zhang. Dept. of Plant Biology and Pathology, Rutgers University, New Brunswick, NJ 08901.

Panicum is a genus in the grass family Poaceae. It contains 400+ species of tall grasses that are native in the tropics, with a few species also distributed in the northern temperate zone. Panicum virgatum, the switchgrass, is a potential bioenergy species. The objective of this study is to explore fungal communities associated with Panicum. In 2011, we sampled cultivated P. virgatum in New Jersey (NJ) and naturally occurring Panicum from New Jersey and Hawaii. Root and shoot tissue was surface sterilized and plated on acidified malt extract agar. Fungi were recovered from 79 of 1600 root and shoot segments and were identified based on morphology and DNA sequences of the internal transcribed spacer (ITS) region of ribosomal RNA genes. Isolation of fungi was twice as high in NJ natural locations than cultivated. The dominant fungal species isolated from the natural NJ locations were Fusarium, Bionectria, and Stagnospora. For cultivated NJ samples, Fusarium, Bionectria, and Cladosporium were the most frequently isolated fungi. The most prevalent species in all locations was Fusarium oxysporum. The impact of the F. oxysporum strain to switchgrass host is under investigation.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P32: Two new hyphomycetes, Strelitziana windsorensis sp. nov. and Zeloasperisporium toonae sp. nov., from Toona sinensis in Connecticut

De-Wei Li1* and Guihua Zhao2. 1The Connecticut Agricultural Experiment Station, Valley Laboratory, 153 Cook Hill Road, Windsor, CT 06095, 2Center of Biotechnology R&D, Jiangsu Polytechnic College of Agriculture and Forestry, 3 Jushu Road, Jurong, Jiangsu 212400, China.

Two new hyphomycetes, Strelitziana windsorensis and Zeloasperisporium toonae, that were collected from Toona sinensis in Windsor, Connecticut were found to be new to science according to the morphological characters and analysis of ITS sequences. Strelitziana windsorensis is described and illustrated with filiform conidia, smooth, 63-103 × 2.0-2.5 μm and conidiophores reduced to conidiogenous cells and Zeloasperisporium toonae with conidia, smooth, 12.3-17 × 3.5-4.4 μm, 0-2 (1) septate. Keys to the species of Strelitziana and Zeloasperisporium are provided.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P33: Molecular, chemical and morphological diversity of Periglandula, clavicipitaceous symbionts of Convolvulaceae (Morning Glories)

Wesley T Beaulieu1*, Wittaya Kaonongbua1, Daniel G Panaccione2, and Keith Clay1. 1Department of Biology, Indiana University, Bloomington, IN 47405, 2Division of Plant and Soil Sciences, West Virginia University, Morgantown, WV 26506.

Fungal endophytes in the Clavicipitaceae (Ascomycota: Hypocreales) have received considerable attention for production of bioactive ergot alkaloids in associations with monocotyledonous plants, primarily economically important grasses. The source of ergot alkaloids in the dicotyledonous Convolvulaceae (morning glories) was unknown until the description of two clavicipitaceous fungi, Periglandula ipomoeae and P. turbinae, which form associations with different host species in the Convolvulaceae. There are dozens of Convolvulaceae known to contain ergot alkaloids, with potentially hundreds more, suggesting each harbors a clavicipitaceous symbiont responsible for alkaloid production. Here we report on a molecular, chemical and morphological survey of six species of Ipomoea (Convolvulaceae) which contain ergot alkaloids for the presence of clavicipitaceous fungal symbionts. All six host species exhibited characteristic epiphytic mycelia on adaxial surfaces of young leaves similar to those of described Periglandula (visualized with SEM) and contained unique profiles of ergot alkaloids in seeds (measured with HPLC). There were considerable intraspecific differences in the density of epiphytic mycelia on young leaves and the presence and composition of ergot alkaloids in foliage. Using PCR and molecular cloning we sequenced two loci from fungi derived from each host: the internal transcribed spacer (ITS) region and the dimethylallyl tryptophan synthase gene (dmaW), which codes for the enzyme that catalyzes the first step in ergot alkaloid biosynthesis. In phylogenetic analyses, fungal ITS sequences from each host species were quite similar except for those from the two most divergent hosts (I. hildebrandtii and I. amnicola) and together formed a clade with described Periglandula spp. Sequences of dmaW were more variable, and sequences from each fungus formed a distinct clade, suggesting they are unique species, except for fungi from the two most closely related species (I. gracilis and I. muelleri), which grouped together. These results suggest there are many species of Periglandula beyond the two described.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P34: Tropical species of Tubeufiaceae (Dothideomycetes) in northern Thailand

Saranyaphat Boonmee* and Amy Y Rossman. Systematic Mycology and Microbiology Laboratory, USDA-ARS, Beltsville, Maryland 20705, USA.

Fifteen species of Tubeufiaceae were found on decaying wood in northern Thailand. Genera in this family are characterized by pseudothecial ascomata that are superficial, light brown, dark brown to black, often smooth. The bitunicate asci are cylindrical to broadly clavate and contain eight ascospores that are filiform, cylindrical to narrowly fusiform, tapering towards rounded to sub-acute ends, multiseptate, hyaline, and pale yellow or brown. Some of these fungi produce a hyphomycetous asexual state in culture such as Chlamydotubeufia khunkornensis with chlamydospores, Thaxteriella inthanonensis with helicoma-like conidia, and Thaxteriellopsis lignicola with helicomyces-like conidia. The relationship between sexual and asexual states is poorly known especially for species from Thailand. Several of these taxa appear to be new to science.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P35: Chemotypic and genotypic diversity in the ergot alkaloid pathway of Aspergillus fumigatus

Sarah L Robinson* and Daniel G Panaccione. West Virginia University, Division of Plant and Soil Sciences, Morgantown, WV 26505.

Aspergillus fumigatus is an opportunistic human pathogen that synthesizes a group of mycotoxins via a branch of the ergot alkaloid pathway. This fungus is globally distributed, and genetic data indicate that isolates recombine freely over that range; however, previous work on ergot alkaloids has focused on a limited number of isolates. We hypothesized that A. fumigatus harbors variation in the chemotype of ergot alkaloids and genotype of the ergot alkaloid gene cluster. Analysis of thirteen isolates by high performance liquid chromatography revealed four distinct ergot alkaloid profiles or chemotypes. Five isolates completed the A. fumigatus branch of the ergot alkaloid pathway to fumigaclavine C. Six independent isolates accumulated fumigaclavine A, the pathway intermediate immediately prior to fumigaclavine C. One isolate accumulated only the early pathway intermediates chanoclavine-I and chanoclavine-I aldehyde, and one isolate lacked ergot alkaloids altogether. A genetic basis for each of the observed chemotypes was obtained either by PCR analysis of the ergot alkaloid gene cluster or through sequencing of easL, the gene encoding the prenyl transferase that reverse prenylates fumigaclavine A to fumigaclavine C. Isolates also exhibited differences in pigmentation and sporulation. The ergot alkaloid chemotypes were widely distributed geographically and among substrate of origin.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P36: Taxonomic and phylogenetic studies of Dothideomycetes on bamboo in Chiang Rai Province, Northern of Thailand

Rungtiwa Phookamsak1*, Amy Y Rossman2, Ekachai Chukeatirote1, and Kevin D Hyde1. 1School of science, Mae Fah Luang University, 57100, Chiang Rai, Thailand., 2Mycology & Microbiology Laboratory, USDA-ARS, Beltsville, MD 20705, USA..

Bamboos are fascinating and useful plants that have a wide range of uses. The traditional people in Southeast Asia, China, Japan, India and South America have used bamboos in their cultures and survival since ancient times. There are several studies on endophytic, pathogenic and saprobic fungi on bamboo. However, phylogenetic information based on molecular data is poorly known for many bambusicolous fungi. We studied the taxonomy and phylogeny of Dothideomycetes on bamboo in Chiang Rai Province, Northern Thailand. Specimens were collected from six locations in Chiang Rai. Pure cultures were obtained using single spore isolation. We identified bitunicate fungi on dead stems or branches of bamboo which belong to the families of Aigialaceae, Didymosphaeriaceae, Lophiostomataceae, Massarinaceae and Melanommataceae. Some fungi are illustrated and a checklist of fungi examined on bamboo is presented. Among the 33 saprobic fungi on dead materials two isolates of Aigialaceae, four of Didymosphaeriaceae, three of Lophiostomataceae and Massarinaceae and 11 of Melanommataceae were identified. The molecular phylogeny of Dothideomycetes on bamboo is inferred based on combined sequences of rDNA, the internal transcribed spacers (ITS rDNA), small subunit nuclear rDNA (18S, SSU), large subunit nuclear rDNA (28S, LSU) and partial RNA polymerase second largest subunit (RPB2).

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P37: Aspergillus section Versicolores: nine new species and multilocus DNA sequence based phylogeny

Stephen W Peterson. 1815 North University Street, USDA, Peoria, IL 61604.

β-tubulin, calmodulin, internal transcribed spacer and partial lsu-rDNA, RNA polymerase, DNA replication licensing factor Mcm7, and pre-rRNA processing protein Tsr1 were amplified and sequenced from 62 A. versicolor clade isolates and analyzed phylogenetically using the concordance model to establish species boundaries. We used phylogeny to define species and phenotype from 10d CYA cultures to describe the species. Aspergillus austroafricanus, A. creber, A. cvjetkovicii, A. fructus, A. jensenii, A. puulaauensis, A. subversicolor, A. tennesseensis and A. venenatus are described as new species and A. amoenus, A. protuberus, A. tabacinus and A. versicolor are accepted as distinct species on the basis of molecular and phenotypic differences. PCR primer pairs used to detect A. versicolor in sick building syndrome studies have a positive reaction for all of the newly described species.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P38: A new species of Aliquandostipite from an urban lagoon of Tabasco, Mexico

Maria C Gonzalez1*, Anthony E Glenn2, Edmundo Rosique-Gil3, Kevin D Hyde4, and Richard T Hanlin5. 1Departamento de Botánica, Instituto de Biología, Universidad Nacional Autónoma de México, Mexico City, DF 04510, Mexico, 2Russell Research Center, Toxicology & Mycotoxin Research Unit, USDA, ARS, Athens, GA 30605, USA, 3División Académica de Ciencias Biológicas, Universidad Autónoma Juárez de Tabasco, Villahermosa Tab 86039, Mexico, 4School of Science, Mae Fah Luang University, Chiang Rai, Thailand, 5Museum of Natural History Annex, University of Georgia, Bogart, GA 30622, USA.

During a continued survey of Ascomycota diversity from different lentic freshwater habitats, we recorded several interesting fungi from a lagoon located in the urbanized area of the Villahermosa City, capital of the State of Tabasco. An uncommon specimen was obtained using a submerged wood-baiting technique. This Mexican microorganism shows some of the typical characteristics of Aliquandostipite type species, A. khaoyaiensis i.e. subglobose, ostiolate ascomata; 8-spored, bitunicate, asci; bi-celled, multiguttulate, sheated ascospores and pigmented hyphae up to 40 µm wide. However, in addition, it has other relevant features such as the presence of very long, subcylindrical, contorted, branched, ascomatal neck that when young is covered with hyaline, straight, attenuated, 20-50 µm × 2-4 µm hairs, a characteristic is not described in Aliquandostipite species. Other characteristics that the new fungus exhibits include different average size of mature ascomata, asci and ascospores, the distinctly bitunicate, thin-walled, deliquescent asci and ascospores deeply constricted at the septum that separate into two part spores. The analysis of morphological set of characters and ITS rDNA sequence data confirmed this Mexican lignicolous fungus belongs to Aliquandostipite, but it is genetically different from all the other described species. Therefore, this fungus is proposed as a new species and is described and illustrated. The high frequency of occurrence of this ascomycete at heavily polluted tropical urban lagoon possibly indicates a successful adaptation to that habitat.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P39: Species richness and distribution patterns of freshwater ascomycetes along an altitudinal gradient in the Peruvian Andes

Steven E Zelski1, Carol A Shearer1*, Huzefa A Raja3, and Andrew N Miller2. 1Department of Plant Biology, University of Illinois, 505 S. Goodwin Ave., Urbana, IL 61801, 2Illinois Natural History Survey, University of Illinois, 1816 South Oak St., Champaign, IL 61820, 3Department of Chemistry and Biochemistry, University of North Carolina Greensboro, 457 Sullivan Science Building, Greensboro, NC 27402-6170.

We sampled freshwater ascomycetes along an altitudinal gradient in the Peruvian Andes to determine their spatial distribution patterns along a climatic gradient. Previous, but as yet unpublished work has shown a near complete turnover of species along a latitudinal gradient reaching from the sub-Arctic to the tropics and one might expect a similar pattern to emerge with elevation, as has been shown for terrestrial plants and aquatic invertebrates. Submerged woody debris was collected from a variety of aquatic habitats including palm swamps, ponds, streams and rivers along with environmental data such as water temperature, pH, dissolved oxygen content and GPS location. For data analyses, sites were categorized as low, medium, or high elevation along an altitudinal gradient ranging from 218-3870 m. Approximately 30 pieces of decomposing plant debris were collected at each site and examined in the laboratory for the presence of fungal fruiting structures. Data for this abstract is based on two collecting trips, one in May and the other in September and October of 2010. Over 1250 individual fungal specimens have been documented, representing nearly 300 species. Preliminary analysis of the distribution data suggests that the three elevational levels differ in their community assemblages and that species turnover occurred along the altitudinal gradient. Species richness was highest at mid elevation (153 species), followed by low elevation (95 species) and high elevation (39 species). Low and mid elevation showed the highest degree of similarity (.481 Sørensen similarity index), while both low and mid elevations showed low similarity to high elevation (.285 and .263 Sørensen similarity index, respectively). Comparison of the Peru species with a summary list of species from several inventories from another tropical region, namely Thailand, reveals a Sørensen similarity index of .152 suggesting there are pantropical species, but differences in longitudinal assemblages do exist.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P40: Environmental sampling as a means of rapid identification of freshwater fungal assemblages on submerged woody debris

Steven E Zelski1*, Andrew N Miller2, and Carol A Shearer1. 1Department of Plant Biology, University of Illinois, 505 South Goodwin Ave., Urbana, IL 61801, 2Illinois Natural History Survey, University of Illinois, 1816 South Oak St., Champaign, IL 61820.

Quickly identifying the members of a fungal community colonizing submerged woody debris is a goal for those interested in studying aquatic fungi. Traditional methods are both time consuming and require expert knowledge of freshwater fungal taxonomy, thus a molecular identification method is desirable to address these issues. While environmental sampling has been done on various other substrates, no previous work has been performed on submerged woody debris. To fill this void a sampling protocol for submerged woody debris was developed to follow changes in the fungal community on submerged balsa wood baits over a seven month time period. Balsa wood baits were attached to bricks and set out in two transects in an Illinois stream. One sample from each transect was retrieved each month of the study and water temperature and pH were recorded. Water temperature ranged from 2.2-19.5 C and pH ranged from 8.0-10.0. Whole community fungal DNA was extracted from the wood samples, the Internal Transcribed Spacer (ITS) of the nuclear rDNA was amplified, clone libraries were constructed , and individual clones were sequenced. Sequences were then sorted into operational taxonomic units and BLAST searches performed in GenBank. In total, 25 taxa were recovered over the course of the study. Species diversity was found to be lower on balsa wood baits than on naturally occurring substrates, which may reflect differences in substrate quality. This study illustrates that not only is it possible to rapidly assess the species present on submerged woody debris, but that there is also the potential to recover a greater diversity of species using this method.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P41: Measuring oomycete biodiversity in aquatic, forest, and agricultural ecosystems: culture-based and metagenomic approaches

Jaime E Blair, Lauren Coffua*, Alison Greidinger, Amy Chabitnoy, and Lauren Cook. Department of Biology, Franklin & Marshall College, Lancaster, PA 17603.

The fungal-like Oomycota have long been studied by mycologists given their osmotrophic lifestyle and habit of hyphal growth. Many species of oomycetes are devastating pathogens, such as the notorious potato late blight agent, Phytophthora infestans, and the cause of rare human pythiosis, Pythium insidiosum; it is not surprising then that most research has focused on host-pathogen interactions, with relatively little study concerning the natural diversity of saprotrophic species. This knowledge gap has important ramifications on our estimates of oomycete biodiversity, as well as on our understanding of the evolutionary history of this important group. The goal of this study is to estimate and compare oomycete biodiversity from several distinct habitats, including aquatic environments, undisturbed forest soils, and highly managed agricultural settings. Our on-going culture-based surveys have relied on various baiting techniques and have revealed a high level of diversity in aquatic and agricultural environments. However, it has been shown that baiting methods can lead to biased estimates of biodiversity as they tend to favor fast-growing organisms and those producing motile zoospores. We have therefore developed a complementary metagenomic approach using massively parallel pyrosequencing to more thoroughly sample species diversity from the different environments. We are currently verifying two markers for analysis, the mitochondrial cytochrome c oxidase subunit 1 (cox1) locus and the nuclear ribosomal RNA internal transcribed spacers (ITS) and large subunit (LSU) region; previous studies have shown that these loci are able to discriminate among closely related species, and provide phylogenetic signal at a number of taxonomic levels. We expect that this combination of culture-based and sequence-based identification methods will enhance our understanding of oomycete ecology and evolution, and perhaps give us more insight into the roles of certain species in the outbreak of disease.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P43: Biological and volatile organic compound diversity in fungal endophytes from Malaysian and Ecuadorian rainforests

Jeffrey Shaw*, Daniel Spakowicz, Cambria Alpha, Brian Dunican, Kaury Kucera, and Scott Strobel. Department of Molecular Biophysics and Biochemistry, Yale University, 260 Whitney Ave., JW Gibbs Room 321, New Haven, CT 06520.

Fungal endophytes comprise a ubiquitous yet understudied source of biological diversity. We have targeted this endophytic diversity for study as part of an effort to identify new secondary metabolite chemistry in microorganisms. The concentration of endophyte diversity has been shown be greatest in tropical rainforests so, through collaborations with the governments of Ecuador and the Malaysian state of Sarawak, we have undertaken collection expeditions to investigate the biological and chemical diversity of endophytes in these regions. The Ecuador expedition is done in conjunction with the Yale "Rainforest Expedition and Laboratory" course which includes 15 undergraduate students. Based on sequencing of the internal transcribed spacer (ITS) region of the ribosomal RNA in our isolates and comparison to GenBank, the endophytes isolated from the rainforests of Ecuador and Malaysia are among the most divergent fungi currently known. Of the isolates collected, 23% of those from Sarawak (36 total) and 18% of those from Ecuador (121 total) have less than 95% identity to any cultured organism. This metric can be used to roughly estimate the number of new species in our collection, although ITS divergence among species of fungi can vary widely. Of the total sample, 72 isolates (approximately 9%) have less than 85% identity to the cultured organisms in GenBank. Isolates from these expeditions were assayed for volatile organic compound production (VOC) by GC/MS. We have identified several unique metabolites produced by these organisms that have not been previously observed. Among these are a series of molecules, produced by several different fungi, identified as nonatetraenes. Other hydrocarbons were produced with a range of carbon chain lengths (C8 - C20) and varying degrees of unsaturation, branching and cyclization. Several of the compounds identified have interesting bioactivity or properties that may make them attractive biofuels.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P44: Foliar endophytic community structure in wild and cultivated stands of Hawaiian 'ohi'a lehua (Metrosideros polymorpha)

Brian A Perry*, Erin Datlof, and Mali‘o Kodis. Biology Department, University of Hawai‘i at Hilo, 200 W. Kawili St., Hilo, HI 96720.

Although associated with all plants that have been investigated, the taxonomic, genetic and functional diversity of endophytic fungi remains undocumented for many regions, including the Hawaiian Islands. Fungal endophytes have been shown to confer such benefits as increased draught tolerance, resistance to pathogens, and anti-herbivory properties. Additionally, fungal endophyte composition has been shown to play a large role in altering vegetation dynamics and plant community composition. Given the potential roles of fungal endophytes, it is clear that a detailed understanding the taxonomic and genetic diversity of these symbionts should be addressed as conservation and management plans for host species are developed. To assess such diversity in a native Hawaiian taxon, we examined foliar fungal endophytic community structure in elevational phenotypes of wild and cultivated Metrosideros polymorpha ('ohi'a lehua) using environmental PCR methods. Preliminary results based on ITS sequence data indicate that low and high elevation populations on Hawai'i Island harbor disparate endophytic communities. Additionally, seeds collected from high and low elevation populations and reared together in a mid-elevation garden harbor endophytic communities similar to those of their parent populations, suggesting strong host selection and/or vertical symbiont transmission. Seeds isolated from cultivated M. polymorpha individuals are currently being examined as potential vectors of endophytic communities similar to those recovered from foliar samples.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P45: Lichen biodiversity in Big Cypress National Preserve: A pilot study

Klara Scharnagl1* and Frederick Seavey2. 1Agroecology Program, Florida International University, Miami, FL 33199, 2South Florida Natural Resources Center, Everglades National Park, Homestead, FL 33034.

This poster outlines a pilot study of a larger project, which intends to survey the overall lichen biodiversity of Big Cypress National Preserve in south Florida. Big Cypress is a unique habitat, containing a variety of niches including cypress domes and hardwood hammocks. A tremendous and diverse amount of flora and fauna depend upon this preserve, in addition to neighboring areas such as Everglades National Park. It is therefore critical to inventory and monitor the changes of biodiversity over time; measuring not only the impacts of global climate change, but also human impacts such as mining, vehicle traffic, dumping and runoff. Lichens have been used since the eighteenth century as indicators of air pollution - but what can their diversity and abundance tell us about the overall health of an ecosystem? How might lichen-lichen, as well as lichen-epiphyte and other interactions impact the ecosystem? What changes can be seen in lichens as Miami receives more and more errant and unseasonable weather [e.g., distribution, photobiont selection, die-offs]? One cannot even start to answer these questions until we know what lichens are there. This project is just the beginning of attempting to use lichens as indicators of ecosystem health, by creating a database of current lichen diversity, abundance, and distribution. Big Cypress Preserve covers over 700,000 acres. Thus for this pilot study, a small area, "RP," with relatively easy access, was selected. [The RP area also has high human impact; a study of lichens as air pollution indicators will be presented at the BICY Science Research Symposium in Fall 2012]. Overall, twelve cypress domes and two hardwood hammocks were surveyed for lichens. Over two hundred specimens were collected, and are being identified via morphology and TLC. The results of this initial biodiversity survey will be presented.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P46: Multilocus phylogenetic analysis of the Xylariaceae: what are the roles of previously unknown endophytic and endolichenic fungi?

Jana M U'Ren* and A Elizabeth Arnold. School of Plant Sciences, University of Arizona, Tucson, AZ, 85721.

The Xylariaceae (Sordariomycetes) comprise one of the largest and most diverse families of filamentous Ascomycota, with at least 73 accepted genera and ca. 800 species. Currently recognized species include many facultative saprotrophs of wood, litter, soil and dung, and a few plant pathogens that cause canker diseases, root rots, and needle blight in agricultural and natural systems. Despite a strong tradition of taxonomic work on Xylariaceae, recent studies illustrate that an enormous amount of undescribed diversity within the family occurs as previously unstudied endophytic and endolichenic fungi. Previous studies have examined the evolutionary relationships between described Xylariaceae species and unknown fungi for a limited number of genera using SSU rDNA and ITS rDNA, but ribosomal genes either lack resolution (i.e., SSU) or are unalignable across diverse taxa (i.e, ITS), precluding robust phylogenetic analyses at the family level. We isolated fungi from the interior of living leaves, senescent leaves, decaying leaves, and lichen thalli in five biotically and geographically diverse locations across North America, and sequenced ITS-partial LSU rDNA for 2062 xylariaceous isolates. Cluster analyses suggested the presence of 94 putative species (based on 95% sequence similarity) and 249 unique genotypes. For one representative of each putative species, we sequenced two protein-coding genes (RPB2 and beta-tubulin), which were aligned with sequence data from 300 described species of Xylariaceae and analyzed using maximum likelihood and Bayesian inference methods. Preliminary analyses indicate that endophytic and endolichenic fungi frequently form novel well-supported clades relative to described species. Some endophytic/endolichenic fungi were found in clades not previously known to contain symbiotrophic taxa. On-going work will pair phylogenetic analyses with ancestral state reconstructions to examine the evolution of major trophic modes and the role of symbiosis in diversification of this important group of fungi.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P47: Cophylogenetic study of the lichen-forming fungus Peltigera and its cyanobiont Nostoc at an intercontinental spatial scale

Eimy Rivas Plata*, François Lutzoni, Orvo Vitikainen, Trevor Goward, Emmanuel Sérusiaux, Nicolas Magain, and Jolanta Miadlikowska. Department of Biology, Duke University. 130 Science Drive, Durham, NC 27708-0338.

The Peltigera aphthosa and P. leucophlebia species complexes are classified within two sections, Peltidea and Chloropeltigera, respectively. These sections include mostly tripartite lichens associated with a green alga (Coccomyxa) as the primary photobiont (in the thallus) and a cyanobacterium (Nostoc) as the secondary photobiont (in the external cephalodia). Only two species, P. malacea and P. frippii, both from section Peltidea, are bipartite with Nostoc as the only photobiont. Members of both sections are restricted to the coldest regions of the northern hemisphere with a peak of species richness in boreal forests. Although P. aphthosa and P. leucophlebia are morphologically similar, to the point of being difficult to distinguish phenotypically, genetically they have diverged extensively with both clades recovered as paraphyletic but never well supported by bootstrap analyses. The phenotypical variation in both clades has triggered the description of several species, such as P. britannica, P. chionophila, P. latiloba, and P. nigripunctata. Using sequence data of the internal transcribed spacer (ITS) and two protein-coding genes (beta-tubulin and RPB1), we inferred phylogenetic relationships for the fungal partner of lichen thalli sampled from North America, Europe, and Asia. Our phylogenies partially support existing species but also suggest a more complex taxonomic structure that requires a reassessment of current species delimitations in these sections. In parallel, we generated a phylogeny for Nostoc from the same thalli using the rbcLx locus, and found a pattern of associations between clades of lichen mycobionts and Nostoc photobionts. In particular, samples of P. malacea, regardless of their geographic origin, appear to be associated with a unique clade of Nostoc, absent in other species, indicating a mycobiont-photobiont cospeciation event during their coevolution. This rare case of cospeciation, could result from a secondary acquisition of the bipartite state, following a transition to a tri-partite state in that section.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P48: Identification of the anamorphs of morels and false morels in Pacific Northwestern U.S. forests

Lori Carris*, Kalyn Thomas, Sean McCotter, and Tobin Peever. Department of Plant Pathology, Washington State University, Pullman WA 99164-6430.

Morels (Morchella), false morels (Gyromitra) and allied genera are widely distributed spring macrofungi in the Pacific Northwestern (PNW) US, and recent studies have supported a western North American origin and diversification for morels. However, relatively little is known about the biology and ecology of this iconic group of fungi, and many parts of the inland PNW remain under sampled. As part of an ongoing study on the diversity and mating systems of black morels (M. elata group) and allied genera, specimens from 15 morel and false morel populations were sampled at three locations in northern Idaho and eastern Washington in May-June 2011. Putative anamorphs were collected at these same sites in Oct-Dec. Anamorph colonies and ascocarps were photographed, morphologically characterized, and established in culture. Brown and white anamorph morphotypes were distinguished based on mycelium color and presence or absence of setae, respectively. Portions of ITS rDNA, RPB1, RPB2, and EF1-alpha were sequenced, and used to confirm anamorph-teleomorph connections for brown anamorphs with black morel lineages, and white anamorphs with Disciotis and Gyromitra spp. This is the first report of anamorphs for Disciotis and Gyromitra, and the first report of the Morchella anamorph from nature. An anamorphic state corresponding to the genus Costantinella has been reported in cultured morels, but nothing is known about the role of the anamorph in the morel life cycle in nature. Identification of the widespread occurrence of the anamorphs of morels and false morels promises to fill major gaps in our understanding of the biology and ecology of these fungi.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P49: Ecological diversification of saddle fungi: Helvella indicated by phylogeny and secondary structure of ITS sequences from annotated collections and environmental samples

Jonathan Hwang1*, Qi Zhao2, Zhuliang Yang2, Zheng Wang1, and Jeffrey P Townsend1. 1Ecology and Evolutionary Biology Department, Yale University, 165 Prospect Street, New Haven, CT 06520, USA, 2Key Laboratory of Biodiversity and Biogeography, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan, China.

Saddle fungi Helvella (Helvellaceae, Pezizomycotina) species are morphologically well-defined and have been intensively studied for their taxonomy and classification. Ecological roles as being saprotrophic or mycorrhizal have been suggested with indirect evidence, including molecular data from environmental samples, for some species in the genus. Recent molecular phylogenies based on internal transcribed spacer of rDNA (ITS) did not support morphological groupings; however, a robust phylogeny of ITS was not achieved due the large divergence observed among Helvella ITS sequences, which makes a dependable alignment of representative ITS sequences in the genus out of reach of traditional alignment programs. In this study, we applied SATé, a software that combines progressive alignment and tree building to build so far the most inclusive ITS alignment for annotated Helvella species and their related environmental ITS sequences. Estimated secondary structures for selected ITS2 sequences verified the robustness of the phylogeny based on the best SATé alignment. Our results suggested a high homoplasious similarity in key morphologies in Helvella, and morphological groupings of the genus do not reflect the true evolutionary histories within the group. ITS phylogeny including environmental samples also provides strong evidence that ectomycorrhizal association is maintained in a clade of Helvella and diversified to adapt for different hosts and geographic conditions. However, saprotrophic life styles in other Helvella species requires expanded metagenomic investigation on plant materials and soil samples, as no single ITS sequence was recovered from studies examining various soil samples for those common fungi.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P50: Helvella vespertina and Helvella dryophila, two new Helvella lacunosa look-alikes from western North America

Nhu H Nguyen1*, Karen Hansen2, Rosanne Healy3, and Else C Vellinga1. 1111 Koshland Hall, Dept. of Plant and Microbial Biology, UC Berkeley, Berkeley CA 94720, USA, 2Swedish Museum of Natural History, Department of Cryptogamic Botany, P.O. Box 50007, SE-104 05 Stockholm, Sweden, 3Department of Plant Biology, University of Minnesota, St. Paul, MN 55108, USA.

Fungal taxonomists have long suspected that the macrofungi of western North America are distinct from eastern North America and Europe. Yet, we continue to apply European species name to western North American species. One such case is Helvella “lacunosa”, a very common ectomycorrhizal species. We collected and sequenced ITS and LSU rRNA genes of Helvella “lacunosa” from California and compared them to European and eastern North American H. lacunosa. Specimens from western North America are distinct from European specimens and from American specimens east of the Rocky Mountains. Furthermore, the western North American “lacunosa” resolved into two species, one occurring with conifers (pines and Douglas fir) and one occurring with hardwood, mostly oak. These two species will be described as Helvella vespertina and Helvella dryophila. Intensive sampling in eastern North America and Mexico is desperately needed to resolve the H. “lacunosa” complex occurring in North America. This effort is a contribution to the North American Mycoflora Project.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P51: Fire drives habitat specialization, speciation, and hybridization of Morel mushrooms in western North America

Todd W Osmundson* and Matteo Garbelotto. Forest Mycology and Pathology Laboratory, Department of Environmental Science, Policy and Management, University of California, Berkeley, CA 94720, USA.

Morels (Morchella spp.) are commercially valuable edible mushrooms and non-timber forest products. Several species reproduce prolifically the year after wildfires, suggesting a response to - and perhaps important role in - post-fire forest regeneration. Although post-fire seed regeneration may accelerate evolutionary diversification compared to adult persistence in plants, similar comparisons for fungi are lacking. Using multilocus phylogenies and diversification analyses, we demonstrate the evolutionary importance of fire-associated reproduction in morels through (i) high ecological specificity of fire association; (ii) phylogenetic conservatism of fire-association, with diversification concurrent with shifts in fire-association status; and (iii) gene introgression between phylogenetically distinct but ecologically similar fire-associated species. Rates of phylogenetic diversification and molecular evolution do not differ between fire- and non-fire-associated lineages, suggesting that rate-enhancing factors (e.g., population turnover, large population sizes) are counteracted by rate-limiting ones (e.g., long fire-return intervals). Genetic introgression was observed only between fire-associates, suggesting that fire can counteract diversification by synchronizing sexual reproduction between species, a process potentially relevant to other species that reproduce in response to disturbance.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P52: What is a cup fungus doing fruiting from a truffle tree? The case of the truffle-cup fungus lineage /pachyphloeus-scabropezia

Rosanne A Healy1*, Gregory M Bonito2, Matthew E Smith3, Gonzalo G Guevara4, Jonathon Frank5, Darlene Southworth5, James M Trappe6, and David J McLaughlin1. 1Department of Plant Biology, University of Minnesota, St. Paul, MN 55108, USA, 2Department of Biology, Duke University, Durham, NC 27708, USA,, 3Department of Plant Pathology, University of Florida, Gainesville FL, USA, 4Instituto Tecnológico de Ciudad Victoria, Biology, Ciudad Victoria, Mexico, 5Department of Biology, Southern Oregon University, Ashland, OR 97520, 6Department of Forest Ecosystems and Society, Oregon State University, Corvalis, OR, USA.

The ectomycorrhizal genera Pachyphloeus, a truffle, and Scabropezia, a cup fungus, are found primarily in broad-leaved forests of the Northern Hemisphere. Respectively they have twelve (plus two varieties) and three accepted described species. Molecular phylogenetic analyses have placed Scabropezia within Pachyphloeus. Previous inferences of truffle evolution in the Pezizales suggest that fruitbody evolution proceeds from cup fungus form to truffle form with no known reversals. Current data is unclear on whether Scabropezia branches from the base of the Pachyphloeus lineage, or is nested within Pachyphloeus, making it paraphyletic. The latter scenario may imply a reversal of form from truffle to cup fungus. Taxonomic confusion in Pachyphloeus from poorly understood type descriptions has resulted in expansion beyond original species limits, which in turn has led to confusion regarding the relationship of Scabropezia to Pachyphloeus. The goals of this study were to revise the /pachyphloeus-scabropezia lineage using multigene analyses from sporocarp and anamorph DNA sequences, and to infer the relationship of Scabropezia to Pachyphloeus. The ITS rDNA was sequenced from representative specimens from across the Northern Hemisphere, and then sorted into phylotypes based on 96% similarity. The LSU, RPB1 and RPB2 loci were sequenced from selected specimens to better resolve relationships within the lineage. Morphological data was mapped onto one of the best-supported maximum likelihood phylograms to identify phylogenetically informative features. Preliminary results suggest that 1) the Scabropezia clade branches from the base of the /pachyphloeus lineage, but includes P. austro-oregonensis, 2) there are at least 25 species of Pachyphloeus and four species of Scabropezia, 3) P. lateritius and P. macrosporus are not part of the lineage, and 4) anamorph color, sporocarp color, ascus shape, and spore wall ornamentation are congruent with molecular data.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P53: The Ecology of Morchella anatolica

Mustafa Isiloglu*, Hakan Alli, and Hayrunisa Bas Sermenli. Mugla University Dept. of Biology, Campus, Mugla, Turkey.

Morchella anatolica Isiloglu, Spooner, Allı & Solak was described in 2010 from very interesting area in Mugla, Turkey. In this paper , its remarkable and unique ecology is discussed.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P54: Phylogeography and diversification of Truffles in the Tuberaceae

Gregory Bonito*, Matthew Smith, Michael Nowak, Rosanne Healy, Gonzalo Guevara, Efren Cázares, Akihiko Kinoshita, Eduardo Nouhra, Laura Domínguez, Leho Tedersoo, Claude Murat, Yun Wang, Baldomero Arroyo Moreno, Donald Pfister, Kazuhide Nara, Alessandra Zambonelli, James Trappe, and Rytas Vilgalys. Deparment of Biology, 125 Science Drive, Duke University, Durham NC 27708, USA.

Truffles have evolved from epigeous (above-ground) ancestors in nearly every major lineage of fleshy fungi. Because accelerated rates of morphological evolution accompany the transition to the truffle form, closely related epigeous ancestors remain unknown for most truffle lineages. This is the case for the quintessential truffle genus Tuber, which includes species with socio-economic importance and esteemed culinary attributes. Ecologically, Tuber spp. form obligate mycorrhizal symbioses with a diverse species of plant hosts including pines, oaks, poplars, orchids, and commercially important trees such as hazelnut and pecan. Unfortunately, limited geographic sampling and inconclusive phylogenetic relationships have obscured our understanding of their origin, biogeography, and diversification. To address this problem, we present a global sampling of Tuberaceae based on DNA sequence data from four loci for phylogenetic inference and molecular dating. Our well-resolved Tuberaceae phylogeny shows high levels of regional and continental endemism. We also identify a previously unknown epigeous member of the Tuberaceae - the South American cup-fungus Nothojafnea thaxteri (Cash) Gamundí. Phylogenetic resolution was further improved through the inclusion of a previously unrecognized Southern Hemisphere sister group of the Tuberaceae. This morphologically diverse assemblage of fungi includes truffle forms endemic to Australia and South America (e.g. Gymnohydnotrya spp.) and non-truffle species. The Southern hemisphere taxa appear to have diverged more recently than the Northern Hemisphere lineages. Our analysis of the Tuberaceae suggests that Tuber evolved from an epigeous ancestor in close association with angiosperm hosts. Molecular dating estimates of ~134 million years ago for Tuberaceae are in accord with this hypothesis. Intra-continental diversification, limited long-distance dispersal, and ecological adaptations have driven evolution and Tuberaceae biodiversity.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P55: Membranomyces species are common ectomycorrhizal symbionts across the northern hemisphere

Jessie K Uehling1*, Terry W Henkel1, Rytas Vilgalys2, and Matthew E Smith2,3. 1Department of Biological Sciences, Humboldt State University, Arcata CA 95521, USA, 2Department of Biology, Duke University, Durham NC 27708, USA, 3Department of Plant Pathology, University of Florida, Gainesville FL 32611, USA.

Membranomyces Jülich is an ectomycorrhizal (ECM) resupinate basidiomycete genus that currently contains two species, M. delectabilis and M. spurius, known from Europe and Canada. Studies have suggested that Membranomyces falls within the Clavulinaceae (Cantharellales), which otherwise consists of the largely coralloid genus Clavulina. Some phylogenetic analyses have resolved Membranomyces as sister to Clavulina, while others suggested that Membranomyces is nested within Clavulina. With the recent discovery of two tiny, resupinate Clavulina species in Guyana, we hypothesized that some of the "missing" Clavulina species diversity indicated by belowground molecular studies in Guyana may be composed other similarly cryptic taxa. Given that a number of temperate belowground ECM studies have found Clavulinaceae root tip sequences that were unidentified at the species level, we suspected that a missing diversity situation might also apply to Membranomyces. We sequenced ITS rDNA from herbarium specimens of M. delectabilis and M. spurius to provide a database from which to determine if conspecific sequences were already present from root or soil samples deposited in GenBank. We found numerous Membranomyces environmental sequences on GenBank; subsequent phylogenetic analyses including these and specimen sequences indicated that 1) Membranomyces species are more widely distributed in the northern hemisphere than previously thought, forming ectomycorrhizas with a diverse array of gymnosperm and angiosperm host plants, and 2) unidentified Membranomyces species exist. Host plant genera with Membranomyces associates included Arctostaphylos, Betula, Carpinus, Fagus, Populus, Quercus, Pinus and Tsuga, with an apparent new distribution including Europe, the Mediterranean Arc, the Middle East, Japan, New Zealand, and United States. We discuss the taxonomic, ecological, and biogeographical implications of these findings.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P56: Host and tissue specificities in Galerina and Mycena species associated with boreal forest mosses revealed by data-mining of amplicon pyrosequencing fungal inventories

Rune Heimdal1, Marie L Davey1,2, and Håvard Kauserud1*. 1Molecular Evolution Research Group, Department of Biology, University of Oslo, N-0316 Oslo, Norway, 2Department of Ecology and Natural Resource Management, Norwegian University of Life Sciences, N-1432 Ås, Norway.

The agaricoid basidiomycete genera Galerina and Mycena are both common saprophytes in the boreal forest that are frequently reported in 'mossy habitats' or in the presence of mosses. ITS2 Sequences with BLAST matches to Galerina and Mycena were extracted from three large scale, amplicon pyrosequencing-based inventories of moss-associated fungi in southern Norway, and used for investigations into the genetic diversity of both groups and their ecological preferences. In both genera, genetic diversity was high relative to reported alpha diversity in those habitats. A relatively small number of OTUs occurred with high frequency and abundance, and in addition, a larger number of taxa were rare and occurred at very low frequencies and abundances. Among those common taxa, most OTUs showed a distinct preference for senescent moss tissues, supporting the idea that these fungi are decomposers of bryophytes. However, a number of OTUs also occurred with high frequency and abundance in photosynthetic tissues, suggesting some species have a facility for colonization and persistence in living moss tissues. In addition, some OTU's showed a distinct predilection for particular moss hosts, suggesting that identifying the moss associated with fruitbodies of these fungi may prove useful in the identification of the fungus.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P57: Systematics of Lactarius in the Rocky Mountain alpine zone

Edward G Barge* and Cathy L Cripps. Plant Sciences and Plant Pathology Department, Montana State University Bozeman MT 59715.

Lactarius is an important ectomycorrhizal genus in the Arctic-Alpine Biome where it associates primarily with Salix and Betula species. The Arctic-Alpine Biome covers roughly 8% of the earth's land and in the Rocky Mountains of North America the alpine is comprised of scattered "islands" above timberline in mountainous areas. Beginning in 1999 Cripps, Horak and others surveyed arctic-alpine macromycete distributions in the Rocky Mountains of Montana, Colorado and Wyoming. Studies of the ectomycorrhizal fungi present in arctic-alpine areas are of importance as climate change impacts this biome, which includes the range expansion of Salix species, a key alpine ectomycorrhizal phytobiont. This study focuses on the systematics of Lactarius in alpine areas of the Rocky Mountains. Macromorphological descriptions made at the time of collection, microscopic examination of dried material and phylogenetic analysis (rough) have contributed to identification. Drawings of spores, pleuromacrocystidia and cheilomacrocystidia were completed for each species using a Leica drawing tube and are complemented with SEM photographs for each species. DNA from upwards of 48 collections was successfully extracted and the ITS region amplified using primers ITS1-F and ITS4. Preliminary analysis shows morphologically identified species grouping together nicely. Thus far, the study has resulted in identification of six alpine Lactarius species: L. glyciosmus, L. lanceolatus, L. nanus, L. pseudouvidus, L. repraesentaneus and L. salicis-reticulatae. Other than a preliminary report, most are first documentations for these species in alpine habitats south of the Canadian border. Host patterns are possible among the species that occur with dwarf and shrub Salix and one species seems to occur only with Betula. These species are known from many arctic-alpine habitats throughout the world in places such as Greenland, Iceland, the Alps, Svalbard and Scandinavia and are now documented for the Rocky Mountains of Montana, Wyoming and Colorado.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P58: Evolutionary relationships of the gomphoid genus Clavariadelphus: One genus or two?

Andrew S Methven1* and Andrew N Miller2. 1Department of Biological Sciences, Eastern Illinois University, Charleston, IL 61920, 2Illinois Natural History Survey, University of Illinois, Champaign, IL 61820.

The genus Clavariadelphus includes a group of club-shaped basidiomes most commonly collected in late summer and fall in northern, boreal forests throughout North America. A monograph of Clavariadelphus in North America divided the genus into two subgenera: subgenus Clavariadelphus which includes ectomycorrhizal species that are associated with coniferous or deciduous trees, broadly ellipsoid basidiospores (length:width ratio < 2.5), and little or no hyphae at the base of the basidiomes; and, subgenus Ligulus with saprotrophic species that function ecologically as litter decomposers in coniferous forests, narrowly ellipsoid basidiospores (length:width ratio > 2.5), and copious amounts of hyphae which bind the substrate to the base of the basidiomes. While some agaricologists have argued that these two subgenera are distinct enough to be recognized as separate genera, questions about the range of variation in morphological characters, chemical spot tests and cultural characters have precluded recognition of the two groups as segregate genera. We hypothesize that the genus Clavariadelphus is polyphyletic and, in order to adhere to a natural system of classification, needs to be subdivided into two monophyletic groups or genera. Two nuclear ribosomal genes, ITS and LSU were amplified, sequenced and analyzed in a phylogenetic context to determine if Clavariadelphus should be segregated into two genera. Based on ITS and LSU sequences, Subgenus Ligulus is well-supported as a monophyletic group that is distinct from and basal to Subgenus Clavariadelphus. Since the type species of the genus, C. pistillaris, belongs to Subgenus Clavariadelphus, a new genus will be proposed for the taxa included in Subgenus Ligulus.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P59: The Boletineae of Queensland, Australia

Mitchell Nuhn*, Roy E Halling, Manfred Binder, David S Hibbett, and Todd W Osmundson. Department of Biology, Clark University, 950 Main St, Worcester, MA 01610.

The Boletineae is a group of mushrooms with many species highly sought after by both animals and humans as food. The majority of Boletineae species are ectomycorrhizal with woody plants, making them important members of forest ecosystems. This study will provide the first biodiversity inventory of the Boletineae in Queensland, Australia, including three of Australia's Biodiversity Hotspots and World Heritage listed sites. To accomplish the inventory, modern molecular and phylogenetic techniques will be used, in combination with traditional taxonomic methods. A sequence dataset of nuclear ribosomal large subunit (nuclsu) and translation elongation factor 1-alpha (tef1) sequences has been generated for the systematic overview of the Boletineae of Queensland. The study has already resulted in one publication, designating a new cosmopolitan genus and at least two species for a previously described species, Sutorius eximius and S. australianses (previously Tylopilus eximius). Further study is ongoing, providing an overview of amorphologically distinct, cosmopolitan taxon, Tylopilus chromapes, that appears to be polyphyletic based on preliminary results. An overview of additional Boletineae species occurring in Queensland, Australia and their relationships in the context of a worldwide phylogeny of Boletineae is in progress.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P60: A molecular analysis of European and North American taxa within the Xerocomus chrysenteron complex and the description of X. redeuilhii Taylor, Eberhardt & Simonini sp. nov.

Andy FS Taylor1,2, Greg W Douhan3*, Allan E Hills4, Giampaolo Simonini5, Manfred Binder6, and Ursula Eberhardt7. 1The James Hutton Institute, Craigiebuckler, Aberdeen, AB15 8QH, Scotland, UK, 2Institute of Biological and Environmental Sciences, University of Aberdeen, Cruickshank Building, St Machar Drive, Aberdeen, AB24 3UU, UK, 3Department of Plant Pathology and Microbiology, University of California, Riverside, CA 92521, USA, 4‘Megera’, Acremead Road, Wheatley, Oxon, OX33 1NZ, England, UK, 5Via Bellaria 8, 42100 Reggio Emilia, Italy, 6Clark University, Biology Department, Lasry Biosciences Center, 950 Main Street Worcester, Massachusetts 01610-1477, U.S.A., 7CBS-KNAW Fungal Biodiversity Centre, Centraalbureau voor Schimmelcultures, P. O. Box 85167, NL-3508 AD UTRECHT, The Netherlands.

The taxonomy within the genus Xerocomus has not been fully resolved and remains controversial. For example, in North America, Smith and Thiers collapsed all species within the genus Xerocomus into the genus Boletus in the 1970's. Nevertheless, Xerocomus is widely recognized as a distinct genus in Europe and phylogenetic analysis based on LSU-rDNA supports that Xerocomus is a distinct genus. Phylogenetic analyses have also revealed that the type species for the genus, Xerocomus subtomentosus, is distinct from other Xerocomus species that have been studied at the molecular level, which suggests that additional genera could be split off. In this paper we examine the taxa in the Xerocomus chrysenteron complex in Europe and North America using rDNA-ITS sequence data. We describe Xerocomus redeuilhii A.F.S. Taylor, Eberhardt & Simonini sp. nov., a widespread species from southern Europe that was formerly considered to be the North American species X. dryophilus (Theirs) Singer. Xerocomus fennicus (Harmaja) Ladurner & Simonini described from Finland is shown to be synonymous with the North American taxon Xerocomus intermedius (Smith & Thiers) Heinemann, Rammeloo &. Rullier and X. marekii Šutara & Skála is shown to be a red form of X. porosporus Imler. Phylogenetic analysis of the complex demonstrated that few taxa are shared between North America and Europe, with a number of lineages within the complex requiring taxonomic re-evaluation in North America.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P61: Vernal fruiting Cortinarii from the mountains of California and the Pacific Northwest

Dimitar Bojantchev. 345 Shipwatch Ln, Hercules, CA 94547.

The mountains of California and Western North America feature a diverse set of Cortinarius species that fruit in the spring, during, or shortly after snowmelt. This vernal-fruiting phenomenon is much more prevalent in the American West than in Europe. Based on several years of intense collecting in California and adjacent states, utilizing morphological and molecular methods, we have become familiar with 26 Cortinarius species that fruit in the spring and early summer. Their subgeneric breakdown is 16 species in subg. Telamonia, 8 in subg. Phlegmacium (including 2 hypogeous forms), 2 in subg. Dermocybe, 1 in sect. Leprocybe and 1 in sect. Anomali. About one third of these species are very rare and so far represented by single collections only. Every season we encounter species that we had not collected before. Almost all of these are new to science and we are in the process of describing them. Some of these vernal fruiting Cortinarii have a broad distribution throughout the West, while others seem to exhibit a degree of endemism as so far they are known only from California. Most of these vernal fruiting species span the clades within genus Cortinarius, except for one group of Telamonia featuring UV active universal veil that form a well-defined clade (/colymbadinus). Only two of the vernal species are known to fruit in the fall. In this session will review the current state of the studies and present the challenges and directions of future research.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P62: Phylogenetic diversity of the ectomycorrhizal genus Cortinarius (Agaricales, Basidiomycota) in the Arctic

Luis N Morgado1*, Manon Neilen1, Machiel E Noordeloos1, D Lee Taylor2, Ina Timling2, and József Geml1. 1Netherlands Centre for Biodiversity Naturalis, Leiden University, P.O. Box 9514, Einsteinweg 2, 2300 RA Leiden, The Netherlands, 2Institute of Arctic Biology, University of Alaska Fairbanks, Fairbanks, AK 99775-7000.

Mycorrhizal associations are abundant and widespread in almost all ecosystems, and c. 80% of land plant species form associations with mycorrhizal fungi. They play a particularly important role in the functioning of terrestrial arctic ecosystems, where arctic plants are highly dependent on mutualistic relationships with mycorrhizal fungi for survival in these nutrient-poor environments. Ectomycorrhiza (ECM) is the predominant mycorrhiza type in arctic and alpine environments, and ECM fungi are crucial for the survival of arctic shrubs (e.g. Betula, Dryas, Salix). Although recent molecular studies have revealed high diversity in arctic ECM communities, the systematic treatment of several arctic ECM genera is still not adequate. Cortinarius is one of the most abundant genera in the Arctic. Here we present preliminary results on the phylogenetic diversity of the genus Cortinarius in the North American and European Arctic. We analyzed internal transcribed spacer (ITS) rDNA sequences from basidiomata and soil samples collected in northern Alaska and Svalbard using likelihood-based phylogenetic methods. We detected at least 28 phylogroups, from which 18 matched sequences from sporocarps deposited in public databases. These included C. umbilicatus, C. biformis, C. delibutus, C. favrei, C. cinnamomeus, C. aureomarginatus, C. urbicus, C. fulvescens, among others. The other 8 represent previously unsequenced taxa that may or may not be newly discovered species. Future investigations will include multi-gene phylogenetics and morphological analyses.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P63: Willows - the arctic connection: biodiversity of arctic-alpine ectomycorrhizal fungi in Salix repens communities in Dutch North Sea sand dunes

Manon Neilen, Machiel E Noordeloos, Luis N Morgado, and Jozsef Geml*. Nationaal Herbarium Nederland, Nederlands Centrum voor Biodiversiteit Naturalis, Universiteit Leiden, P.O. Box 9514, 2300RA Leiden, The Netherlands.

Dwarf willow (Salix) species are ubiquitous in the Arctic and are the primary hosts for the majority of arctic ectomycorrhizal (ECM) fungi. In Western Europe, the sand dune communities of creeping willow (S. repens) are known to harbour a rich mycoflora and are highly important for nature conservation, water resource management, and recreational purposes. In this project, we assess the biodiversity of ECM fungi in the Arctic (Alaska and Svalbard) and in Dutch coastal dune communities. We generated DNA sequence data from both soil and fruitbody samples and conducted phylogenetic analyses for the major Salix-symbiont ECM genera, such as Cortinarius, Russula, Hebeloma, Inocybe and Tomentella. Our results show that the majority of the ECM basidiomycete species found in soil samples taken from S. repens beds in the Dutch maritime sand dunes also occur in various arctic tundra communities. When comparing our results with long-term records of above-ground diversity (i.e., sporocarp mapping data generated by the Werkgroep Paddenstoelenkartering Nederland, WPN), we found that many of these 'arctic' species are considered rare and/or threatened, and therefore, red-listed in the Netherlands. In addition, we could confidently identify several OTUs to arctic species that had never been reported from the Netherlands before, e.g., Cortinarius favrei, Cortinarius laetissimus, Inocybe straminipes, Russula nana. Based on the many similarities in the ECM communities in arctic tundra and maritime sand dunes, it appears that, in the temperate zone, S. repens may serve as an ecological analog of arctic dwarf willows. In coastal dunes, S. repens is among the very few ECM hosts and its presence strongly enhances fungal biodiversity in these areas. Knowledge on ECM biodiversity can provide insight into the functioning and sustainability of these ecosystems.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P64: Mycorrhizal community composition of Quercus oleoides as a function of stand maturity in a regenerating dry tropical forest

Nikhilesh S Desai. Chicago Botanic Garden, Plant Science Center, 1000 Lake Cook Rd, Glencoe, IL 60022.

Species rich dry tropical forests (DTFs) are the most endangered ecosystems in the tropical biome, threatened by many anthropogenic activities. In Costa Rica, deforestation of the DTF has occurred for centuries; however, conservation efforts over the past 30 years have restored these forests to 47.9% of their original extent. To better understand the role of mycorrhizal fungi in the regeneration of DTFs, this project studies the mycorrhizal communities associated with a younger (10 years old) and older (25 years old) stand of Quercus oleoides (tropical live oak). The study site is Sector Santa Rosa of the Área de Conservación Guanacaste (ACG) in northwestern Costa Rica, where Q. oleoides were once regionally dominant. This project is intended to evaluate: 1) the ectomycorrhizal (EM) diversity in older and younger stands of oak and the overlap in community composition between stands; 2) whether older stands contain more rare EM species than younger stands; and 3) how the balance between arbuscular mycorrhizal (AM) and EM abundance reflects stand age. Data collection occurred over two weeks in July 2011. Using two 20-m x 50-m plots from young and old stands, soil cores were collected and the associated EM root tips were pooled for analysis of species composition using molecular methods. Sporocarps were collected for DNA analysis of fresh fungal tissue. DNA sequences were used to identify EM fungi using BLAST searches against GenBank and UNITE databases. Oak roots without mantles were analyzed for AM presence by staining with Trypan blue. More EM root tip colonization was observed in soil cores from older stands. Preliminary RFLP results suggest that this reflects a greater EM richness in these communities. Further results and discussion will be presented in the poster.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P65: The use of native ectomycorrhizal fungi in the restoration of whitebark pine

Erin R Lonergan* and Cathy L Cripps. Plant Sciences and Plant Pathology Department, Montana State University Bozeman MT 59715.

Whitebark pine is a keystone species in high elevation ecosystems of Western North America; it is rapidly declining throughout much of its range due to blister rust, a mountain pine beetle epidemic and fire suppression. The U.S. Fish and Wildlife Service found it warranted but currently precluded for protection under the Endangered Species Act. Huge restoration efforts are on going with over 200,000 nursery-grown seedlings planted in the Western U.S.; however, seedling survival rates are low. Ectomycorrhizal fungi can enhance conifer survival after out-planting, but previous to this research, whitebark pine seedlings were not inoculated before out-planting. Rhizopogon is often used for inoculation, but we found Suillus species specific to five needle pines to be important ectomycorrhizal partners for young and mature whitebark pine trees. Suillus species are also used for inoculation of cembran pines in Europe. Here the effects of inoculation of whitebark pine seedlings with native strains of Suillus sibiricus were investigated in greenhouse and field studies. In the greenhouse, lower levels of fertilization resulted in a greater abundance of ectomycorrhizae, although all treatments with low N fertilizer allowed for colonization. Long containers promoted colonization, while short containers appeared to promote root disease. The field study in Waterton Lakes National Park, Alberta, Canada, is examining the survival of out-planted whitebark seedlings inoculated with native ectomycorrhizal fungi (Suillus sibiricus). A thousand seedlings were planted in "clusters" to mimic the planting strategy of Clark's nutcracker. Each cluster included one, two, or three inoculated seedlings (in addition to non-inoculated controls). Clusters were planted in burned and un-burned areas, with and without beargrass. First year survival of seedlings was high with 95% of seedlings surviving regardless of treatment or habitat type. Monitoring in 2012 will examine survival rates in the second year when higher mortality rates are likely to occur.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P66: From trees to grasses: Arbuscular mycorrhizal fungi community is shaped by host plant preference

Ko-Hsuan Chen1*, Gregory Bonito1, Hannah Reynolds1, Chris Schadt2, and Rytas Vilgalys1. 1Dept. of Biology, Duke University Box 90338, Durham, 27708, 2Oak Ridge National Laboratory, Department of Energy, Oak Ridge TN 37831.

Arbuscular Mycorrhizal fungi (AMF) are important to terrestrial ecosystems; however, most of AMF community studies focused on AMF associate with grasses rather than trees. We examined the AMF community on a forest tree, Populus deltoides, which is associated with ectomycorrhizal fungi as well as AMF. Soil samples from different localities (North Carolina and Tennessee) were used to inoculate P. deltoides cuttings. Mycorrhization of P. deltoides roots was characterized using 454 pyrosequencing and cloning of the AMF SSU rRNA genes. Subsequently, fungal communities from P. deltoides roots were transferred to Sorghum bicolor, a common plant host used for retaining AMF in pot cultures. To detect AMF community associated with S. bicolor, spores were extracted and identified based on PCR amplification of the SSU rRNA gene using several different primer sets. We compared data generated by these three approaches (1) P. deltoides roots/ 454 pyrosequencing; (2) P. deltoides roots/cloning and Sanger sequencing; (3) S. bicolor roots and rhizosphere /PCR. Each method recovered 35, 10 and 10 OTUs, respectively, but only 2 common OTUs were detected by all three methods. Combined results reveal the dominance of Glomerales and Paraglomerales in the P. deltoides AMF community, with few (less than 0.01%) of the fungal OTUs detected from the Diversisporales and none from Archaeosporales. P. deltoides appears to be preferably mycorrhized by Glomerales and Paraglomerales. Several Rhizophagus OTUs were only detected by S. bicolor/PCR method, suggesting Rhizophagus fungi might exist in Populus roots in very low numbers, but are favored by S. bicolor once this host is available. These results suggest that Sorghum may not always be an optimal alternate host for culturing AMF fungi, particularly those that preferentially associate with Populus. Currently we are investigating the transferability of AMF communities from S. sorghum back to P. deltoides.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P67: Improvement in colonization and seedling survival of Wyoming big sagebrush following inoculation with native arbuscular mycorrhizal fungi

Bill E Davidson* and Marcelo D Serpe. 1910 University Drive, Department of Biological Sciences, Boise State University, Boise, ID 83725-1515.

Inoculation of seedlings with arbuscular mycorrhizal fungi (AMF) is a common practice aimed at improving seedling establishment. The success of this practice largely depends on the ability of the inoculum to colonize the growing root system after transplanting. These events were investigated in Wyoming big sagebrush (Artemisia tridentata ssp. wyomingensis) seedlings inoculated with native AMF. Seedlings were first grown in a greenhouse in 50 mL containers containing sterilized pot cultures (control seedlings) or pot cultures having a mixture of seven mycorrhizal phylotypes (inoculated seedlings). In early spring, three- month old seedlings were transplanted to 24 L pots filled with soil collected from a sagebrush habitat and grown under natural climatic conditions. At the time of transplanting the percent colonization was 0.3 and 57% for control and inoculated seedlings, respectively. Two and a half and five months after transplanting, the percent colonization remained higher in inoculated than in control seedlings with values of about 48 and 20%, respectively. These differences in colonization were associated with differences in seedling survival, which was 24% higher in inoculated than control seedlings. In contrast, biomass per plant and the shoot over root ratio were similar for both treatments. Overall, our results indicate that the inoculum contributed to the colonization of the roots that developed after transplanting resulting in higher levels of colonization than those naturally occurring in the soil. The increased colonization by mycorrhizal fungi may have led to an increase in seedling survival.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P68: Mycorrhizal associations in restored and invaded grasslands

Mia R Maltz1*, Kathleen Treseder1, and Jutta Burger2. 1Department of Ecology and Evolutionary Biology, University of California, Irvine, CA, 2Irvine Ranch Conservancy.

Mycorrhizal fungi perform a variety of ecosystem services, including facilitating plant establishment and improving nutrient uptake by plants. However, little is known about which Southern Californian mycorrhizal fungi facilitate native plant biodiversity in grasslands invaded by Brassica nigra. To compare among restoration sites, soil samples were taken from intact native grasslands (>50% native), manually managed B. nigra plots (30-50% native) highly invaded B. nigra cover (<10-15% native), and introduced B. nigra experimental plots. We predicted that B. nigra will inhibit arbuscular mycorrhizal (AMF) colonization of both native forb and grass roots and that introduced B. nigra plots will have the highest AMF community turnover between the dry and wet seasons. These soils were used to assay the AMF community by "baiting" the fungi with native grasses and forbs. After 45 days, the bioassays from each species and soil type were harvested. We extracted and morphotyped AMF spores from each host plant and soil type. Aboveground and belowground plant biomass and shoot height measurements were recorded. Plant roots were stained to assess AMF percent colonization in grass and forb roots. Comparisons among the extant AMF community across these experimental restoration sites were conducted by molecular analyses. Native grass roots were significantly more colonized by AMF than native forb roots. Across all treatments, Stipa pulchra roots were colonized by significantly more AMF than the forb Plantago erecta and produced significantly more above ground biomass than was produced by P. erecta. Soil hyphae were extracted, analyzed, quantified, and compared among each soil type. In addition, wet and dry seasonal monitoring and fungal trait cataloging may inform future research into the role of these native AMF taxa in providing ecosystem services such as assisting plant establishment in inhospitable environments, drought tolerance, and grassland soil stabilization.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P69: Analyzing the role of white-rot fungi in bioremediation using transcriptome and degradation assays following growth on Number 6 fuel oil

Darcy Young1*, Rachael Martin1, James Rice2, Dimitrios Floudas1, Igor V Grigoriev3, and David S Hibbett1. 1Biology Department, Clark University, Worcester, MA, 01610, 2School of Engineering, Brown University, Providence, RI, 02912, 3US Department of Energy Joint Genome Institute, Walnut Creek, CA, 94598.

The extracellular enzymes that white-rot fungi secrete during lignin decay have been proposed as promising agents for oxidizing persistent pollutants like polycyclic aromatic hydrocarbons, synthetic dyes, explosives, and pesticides. We investigated the abilities of the white-rot fungi Punctularia strigoso-zonata, Irpex lacteus, Trichaptum biforme, Phlebia brevispora, Trametes versicolor, and Pleurotus ostreatus to degrade Number 6 fuel oil in wood sawdust cultures. Our goals are to advise the bioremediation efforts underway at a brownfield redevelopment site on the Blackstone River in Grafton, Massachusetts and to contribute to a broader understanding of the mechanisms of decay in white-rot fungi. We have preliminary results from gas chromatography-mass spectrometry analyses suggesting that Irpex lacteus degrades a significant portion of the compounds in No. 6 oil. Punctularia strigoso-zonata is also the subject of transcriptome profiling to determine if there are genes that are differentially regulated during growth in the presence of No. 6 oil on two different wood substrates. We plan to characterize the genes in the four sequenced transcriptomes using the unpublished annotated genome of P. strigoso-zonata. A clearer understanding of the decay mechanisms of P. strigoso-zonata could streamline the use of other white-rot fungi in bioremediation.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P70: Does ECM induce a clean sweep of decay related genes in Agaricomycetes?

Laszlo G Nagy1*, Dimitrios Floudas1, Manfred Binder1, Igor Grigoriev2, Francis Martin3, and David S Hibbett1. 1Biology Department, Clark University, 2Fungal Genomics Program Lead, DOE Joint Genome Institute, 3Plant Biology Department of Nancy University.

Ectomycorrhizae (ECM) are symbiotic interactions between fungi and green plants with an enormous impact on many ecosystems. The ECM association can be found in several lineages of both the Agaricomycetes and the Viridiplantae, with most of the species found in the Agaricales and Boletales in mushrooms and the Pinales the Rosidae in plants. Macroevolutionary aspects of the origin of ECM formation have been studied extensively by phylogenetic comparative methods, resulting in highly discordant estimates regarding the number of gains and losses throughout the Agaricomycetes, however, its genetic basis is very poorly known. Recent and preliminary results suggest that the evolution of decay-related gene families show spectacular patterns in switches between different nutritional modes of fungi. Therefore, in this study, we set out to record gene family expansions and contractions in selected ECM and saprotrophic taxa and correlate these changes with switches in nutritional mode. Gene phylogenies and comparisons of gene copy numbers reinforce predictions based on single genomes. Of the 9 gene families studied, GH6, GH7, GH61, GH12 and fPOX show considerable loss of gene copies in ECM lineages, whereas gene copy numbers of the Multicopper peroxidase, Dye decolorizing peroxidase, GH5 and GH28 gene families are comparable to those in white-rot and brown rot lineages. This suggests that while some of the decay-related gene families tend to be released from selection in mycorrhizal lineages, eventually leading to extensive gene loss over longer timescales, others retain a function, presumably because these gene families confer more general functions and/or ECM lineages maintain some capability to attack wood.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P71: Transcriptional activity of "GH61"-encoding genes from various fungal species grown on straws

Ingo Morgenstern1,2*, Justin Powlowski1,3, and Adrian Tsang1,2. 1Centre for Structural and Functional Genomics, Concordia University, 7141 Sherbrooke Street West, Montreal, Quebec, Canada, 2Department of Biology, Concordia University, 3Department of Chemistry and Biochemistry, Concordia University.

The efficient degradation of lignocellulosic biomass using microbial enzymes is considered a key step for the production of second generation biofuel and value-added by-products. Efforts to improve commercial cellulase mixtures include the spiking with auxiliary enzymes. Recently, members of the so-called glycosyl hydrolase family 61 (GH61) have attracted much interest due to their cellulase-enhancing properties in enzyme mixtures. Copies for GH61 encoding genes are present in the majority of fungal genomes; however, the copy number of GH61s differs tremendously among the investigated genomes, exceeding thirty copies in some species. Currently, it is not known whether this is merely an example of high redundancy or how far functional differences are present among GH61 paralogs. We are examining the transcriptional profile of "GH61s" both from ascomycete and basidiomycete species grown on alfalfa and/or barley straw as substrate and are interpreting the results in a phylogenetic context.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P72: Degradative enzyme production: a function of phylogenetic or niche relatedness?

Jenny Talbot, Lotus Lofgren*, and Kabir Peay. University of Minnesota Plant Pathology 1991 Upper Buford Circle 495 Borlaug Hall St. Paul, MN 55108-6030A.

Fungi produce an array of extracellular enzymes that assist in the degradation of organic materials. Fungi occupying different ecological niches are expected to have different enzymatic capabilities but the extent to which these capabilities are restricted by genetic lineage is unclear. We are testing the hypothesis that fungi occupying similar ecological niches have similar enzyme profiles but that this capability is restricted by genetic lineage. To test this hypothesis, we are using 18 sets of closely related fungi across the Basidiomycota in a laboratory-based microcosm study to directly quantify the production of degradative enzymes among functionally and phylogenetically distinct taxa. Each set consists of 'three pairs' of fungi occupying distinct ecological niches: a mycorrhizal fungus, a white rot fungus, and a brown rot fungus, for a total of 54 taxa. We are measuring the production of cellulases, hemicellulases, proteases, and oxidases by each taxa as they colonize litter and soil in axenic culture. We will then correlate genetic distance to functional distance among taxa and test the effect of ecological niche on enzyme production using ANOVA. ECM fungi form associations with host plants under which fungal-sequestered nutrients are exchanged for plant-fixed carbon. By contrast, white rot fungi and brown rot fungi acquire carbon from detritus, with white rot fungi having stronger lignin degrading capabilities than brown rot. Viewing enzyme production as an adaptive functional trait, we expect that the enzymatic profiles of different fungal species will vary most strongly with ecological niche, with a lesser effect of phylogenetic distance between lineages. However, we expect that closely related species will produce a similar suite of enzymes.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P73: Insights into the potential diversity of the brown rot decay mechanism provided by substrate compositional analysis

Justin T Kaffenberger* and Jonathan S Schilling. Department of Bioproducts and Biosystems Engineering, University of Minnesota, 2004 Folwell Avenue, St. Paul, MN 55108.

Wood-degrading fungi are typically designated by the type of decay that they produce: the two main types being brown rot and white rot. While much effort has been put into understanding the biochemical mechanism by which brown-rot occurs, the process remains poorly understood. Evidence suggests that brown-rot fungi incorporate a Fenton reaction-based oxidation to rapidly de-polymerize substrate components and several mechanisms incorporating Fenton chemistry have been proposed. These proposed mechanisms have been based largely on studies performed on only a few brown-rot species. The polyphyletic nature of brown-rot, however, suggests that there may be several routes to brown rot decay. While these fungi all ultimately produce brown decay residues, details regarding the chemical composition of these residues over the course of degradation have not been explored. Details regarding this chemistry may elucidate similarities and differences in the nature of underlying decay mechanisms. To this end, one representative from each of the known seven clades of brown rot fungi (Gloeophyllum trabeum, Fomitopsis pinicola, Ossicaulis lignatilis, Serpula lacrymans, Fistulina hepatica, Wolfiporia cocos, and Dacryopinax sp.) were used to decay representative hardwood (Populus tremuloides), softwood (Pinus radiata), and grass (Zea mays) species. Compositional analyses of the residues were performed at six time points over the course of 4 months of exposure. Details of these compositions and their implications are discussed.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P74: Spatial mapping of gene expression in the brown rot fungus Postia placenta during wood decay

Gerry N Presley* and Jonathan Schilling. Department of Bioproducts and Biosystems Engineering, University of Minnesota, St. Paul, MN 55108.

Wood is an abundant, renewable feedstock for the production biofuels and biomaterials, however it is very recalcitrant and obtaining fermentable sugars from it and other lignocellulosic material is energetically costly. Brown rot fungi are able to obtain carbohydrates from wood at low temperatures by using a unique mechanism that is thought to involve the production of hydroxyl radicals that modify lignin and allow endoglucanases to access to cellulose in wood cell walls to aid in the release of glucose. Analysis of spruce wafers degraded by Postia placenta suggest that the fungus pretreats its substrate ahead of the advancing hyphal front with oxidative, lignin-modifying chemical reactions followed by saccharification of the substrate, indicated by an increase in endoglucanase activity behind the hyphal front. This project will focus on determining spatial gene expression patterns along a spatial gradient in wood degraded by Postia placenta. Wood sections taken along the length of degrading spruce wafers will be analyzed using Fluorescence in situ hybridization to determine where mRNA from genes thought to be involved in oxidative pretreatment such as quinone reductases and saccharification related genes such as endoglucanases are localized in the degrading wafer. Further analysis of gene expression will be done using qRT-PCR to determine the levels of gene expression for pretreatment and saccharification related genes along the length of degrading spruce wafers. This work will spatially map the expression of genes involved brown rot pretreatment and saccharification and help determine which genes are important in mediating those processes.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P75: Changes in hyphal growth and ultrastructure in two chitin synthase mutants of Neurospora crassa

Karen E Fisher*, Ricardo Reyes, David Lowry, and Robert W Roberson. School of Life Sciences, Arizona State University, Tempe, AZ 85287.

The primary fibrillar component of the fungal cell wall is chitin; a linear polymer of ß-1,4-N-acetylglucosamine. Chitin synthesis occurs at the cell surface, primarily at sites of growth, and is catalyzed by a trans-membrane enzyme known as chitin synthase (CHS). Seven CHSs have been identified in Neurospora crassa. Previous research utilizing GFP fusions of CHS-1, CHS-3 and CHS-6 has documented their accumulation in the core of the Spitzenkörper (Spk) in growing hyphae of N. crassa. The translocation of these enzymes, presumably by vesicle motility, towards the Spk from subapical locations has also been reported. Given the importance of chitin synthesis in fungal cell growth and morphogenesis, we have conducted a study of two chitin synthase deletion mutants in N. crassa: ΔCHS-1 and ΔCHS-6. In this presentation, hyphal growth behavior will be described in these mutants relative to wild-type hyphae. In addition, ultrastructural analysis of cytoplasmic organization will be documented.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P76: Actin dynamics in constricting ring traps formed by nematophagous fungi

Yunluan Cui* and Nicholas P Money. Miami University 316 Pearson Hall Oxford Ohio 45056.

The microfilament network of F-actin plays an essential role in fungal morphogenesis by coordinating cell wall synthesis, cytoplasmic migration, and organelle positioning. We are investigating the connection between the behavior of the actin cytoskeleton and trap development in nematophagous fungi that form constricting ring traps. We have constructed a fluorescence-tagged actin-binding protein (Lifeact) driven by the ToxA promoter through recombinant PCR. The vector pBChygro harboring an ampicillin resistant gene and hygromycin B phosphotransferase gene was used as a backbone to accept the chimeric gene into multiple clone sites through enzyme digestion and linkage. Restriction enzyme mediated integration (REMI) and the CaCl2-polyethylene glycol methods were used to transform fungal protoplasts with the Lifeact-expressing plasmid. This efficient transformation system facilitated the random insertion of the HindIII linearized plasmid into the fungal genome. Transformed protoplasts were imaged using fluorescence microscopy. Treatment with restriction enzymes had no effect on protoplast survival and hyphae generated from protoplasts expressed the fluorescent protein tag. This indicates that heterogeneous Lifeact protein is expressed under the ToxA promoter, and that integration of the plasmid into the fungal genome was successful. This is the first report of the successful transformation of a fungus that produces ring traps using heterogeneous Lifeact by REMI. Data obtained from these experiments will be helpful in the future investigation of functional genes contributing to trap development.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P77: Colorimetric method for discriminating abnormal strains of Flammulina velutipes and SSR markers linked to the defective symptoms

Hee Jeong Je*, Chak Han Im, Na Hee Kwon, Mi Jin Kim, Min Keun Kim, Ki Kwan Park, and Jae San Ryu. Eco-friendliness Research Department, Gyeongsangnam-do Agricultural Research and Extension Services, Jinju 660-360, Republic of Korea.

Winter mushroom, F. velutipes is one of the most popular mushrooms in Asian countries, including Korea, Japan, and China. The strain is preserved on slants or under mineral oil in mushroom farms, but occasionally it degenerates by unknown reasons, resulting in loss in yield and malformation. Therefore, a method is needed that is easy enough to be managed by farmers to detect abnormal strains using a pH indicator and carbon source. Media including MCM, PDA, YP supplemented with 0.5% of carbon sources (sawdust, lactose Avicel, CMC, etc) and pH indicators (bromothymol blue, congo red, methyl orange, bromocresol green, methyl red, bromocresol purple, phenol red, and bromophenol blue) were tested for addressing difference between normality and abnormality of strains. YP supplemented popular sawdust and BTB was useful to detect 2 highly abnormal and 6 slightly abnormal strains which were undetectable by the previous method. 275 SSRs were identified to design primers and to develop markers linked to the defective symptoms. 2 SSR markers showing a polymorphism between normal and abnormal strains were screened. The difference in length of bands was just 4 - 8 bases, so an automatic electrophoresis system will be required to detect such a small difference.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P78: Antifungal effects of Aloe striata on early hyphal development in Fusarium oxysporum and Fusarium solani

Gloria A Wada1,2*, Richard L Bretz2,3, Richard E Edelmann3, and Marcia R Lee1,2. 1Department of Microbiology, Miami University, Oxford, OH 45056, 2Department of Chemistry and Biochemistry, Miami University, Oxford, OH 45056, 3Center for Advanced Microscopy & Imaging, Miami University, Oxford, OH 45056.

Members of the Fusarium solani and Fusarium oxysporum species complexes are the most implicated etiologic agents in opportunistic fusarial infections in mammals. Prevention and treatment of these mycoses are problematic, because the available antimycotics are limited and often have toxic side effects. Popular folk medicines, such as leaf gel from Aloe spp., offer potential sources for novel antimycotic compounds. To screen for antifungal properties of a non-domesticated Aloe spp., specifically Aloe striata, germination assays with homogenized, 0.2 µm filtered A. striata inner leaf gel were performed against conidia of five strains each of F. solani and F. oxysporum. Although exposure to A. striata inner leaf gel caused only minimal inhibition of conidial germination of both F. solani and F. oxysporum strains, it caused notable hyphal aberrations characterized by increased hyphal diameters as well as increased parent cell diameters. To isolate and identify the compound(s) responsible for these aberrations, a combination of chromatographic techniques in addition to IR, LC/MS, 1H and 13C NMR was used. Only one HPLC fraction, specifically fraction A, demonstrated significant induction of hyphal aberrations. Work is in progress to identify the compound(s) within fraction A responsible for the antimycotic activity of A. striata.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P79: Microscopic analysis of Cordyceps bassiana (anamorph stage: Beauveria bassiana) stromata during artificial cultivation for commercial Use

Seong Hwan Kim1, Hyuk Woo Kwon1*, Min Woo Hyun1, Gi-Ho Sung2, and Hyung-Kyoon Choi3. 1Department of Microbiology, Dankook University, Cheonan, Chungnam 330-714, Korea, 22Mushroom Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, Suwon 441-707, Korea, 3College of Pharmacy, Chung-Ang University, Seoul 156-756, Korea.

Cordyceps bassianais is an insect-borne medicinal fungus that has recently been cultivated artificially. To determine the proper time for harvesting fruit bodies of C. bassiana with commercial value, stereo and scanning electron microscopic images of perithecium formation on stomata were analyzed during 13 weeks of artificial cultivation. Perithecia formation initiated on the stromata after 5 weeks in culture. After 6 weeks they were nearly mature on the stromata but ascospores had not been released. Fully matured perithecia producing ascospores were found on the stromata after 7 weeks. The best looking morphological features of fruit bodies with good commercial value were found after 6 to 7 weeks culture. The highest level of total phenolics content was obtained from fruit bodies after 6 weeks culture. Our work suggests the formation and degree of perithecia maturity could be determined according to fruit body development and applied as a criterion for harvest time determination in the production of secondary metabolites from artificially grown C. bassiana stromata.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P80: Characteristics of a new mid-high temperature adaptable oyster mushroom variety 'Gonji-5ho' for bag culture

Jong-In Choi1*, Jeon Dae-Hoon1, Tai-Moon Ha1, and Young-Cheul Ju2. 1430-8, Sam-ri Gonjiam-Eup Gwangju-Si, Gyeonggi-Do 464-873 Repblic of Korea, 2Korea National College of Agriculture and Fisheries, 11-1 Donghwa-Ri Bongdam-Eup Hwaseong-Si Kyeonggi-Do 445-760, Republic of Korea..

'Gonji-5ho,' a new variety of oyster mushroom which is suitable for production in bag culture, was bred by mating between monokaryons isolated from chiak-3ho and Suhan-1ho. The major characteristics of the fruit body included a thick and gray pileus and thick, long and soft stipes. Tissue elasticity and cohesivness was superior to Suhan-1ho. The optimum temperature for mycelial growth was around 26 to 29C and that for pinheading and growth of fruitbodies was around 18 to 20C. In the bag culture, it 18 days of incubation was required and 3 days for the formation of primordia. The fruit bodies grew to maturity and were uniform in size, yielding 221.4g/1kg bag.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P81: Fungal profiles in various milk thistle botanicals from the US market

V H Tournas1*, J Rivera Calo2, and C Sapp3. 1Center for Food Safety and Applied Nutrition/FDA, 5100 Paint Branch Parkway, College Park, MD, USA, 2University of Arkansas, Fayetteville, AR, USA, 3Johns Hopkins School of Medicine, Baltimore, MD, USA.

Milk thistle (MT) dietary supplements are widely consumed due to their possible beneficial effect on liver health. As botanicals, they can be contaminated with a variety of fungi and their secondary metabolites, mycotoxins. This study was conducted in an effort to determine the mycological levels and profiles in various MT botanical supplements (seeds, herb, tea bags, liquid seed extracts, capsules and soft gels) obtained from U.S. retail. Conventional plating methods were used for the isolation and enumeration of fungi, while conventional microscopy as well as molecular methods were employed for species-level identification of the isolated fungal strains. Results showed that a high percentage (62%) of the MT samples tested were contaminated with fungi. Total yeast and mould (Y&M) counts ranged between <100 and 4.0 x 105 colony forming units per gram (cfu/g). MT seeds carried the highest fungal levels followed by MT herb. No live fungi were recovered from MT liquid seed extracts, capsules, soft gels or tea bags. Potentially toxigenic moulds from the Aspergillus flavus and A. niger groups as well as Eurotium, Penicillium, Fusarium and Alternaria species were isolated from MT supplements. The predominant moulds were A. flavus, Eurotium spp. (E. repens, E. amstelodami and E. rubrum), A. tubingensis, A. niger and A. candidus. To our knowledge, this is the first study reporting on fungal contamination profiles of MT botanicals.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P82: Utility of Albasia sawdust for cultivating Pleurotus ostreatus in bottles

Young-Cheol Ju*, Yun-Hae Lee, and Myoung-Jun Jang. Mushroom Research Institute, Gyeonggido Agricultural Research & Extension Services 430-8 Sam-Ri Silchon-Eup Gwangju-Si Gyeonggi-Do, Postal code : 464-873.

In this study, we attempted to find substitute materials for cottonwood sawdust used in the cultivation of Pleurotus ostreatus in bottles. Chemical characteristics of mixed substrates substituting albasia sawdust for cottonwood sawdust were not different significantly. Tests for 184 pesticides (Acrinathrin, etc.) showed that none were detected in the sawdust. The incubation period was shorter using 50% albasia sawdust compared to 100% albasia sawdust. The yield of fruit-bodies was similar to the control and those obtained with 50% albasia sawdust. However, the 50% albasia sawdust treatment was higher in bio-efficiency than the control. Therefore, it is suggested that the 50% albasia sawdust treatment be substituted for cottonwood sawdust for cultivation of P. ostreatus.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P83: Join the dark side: Temptations of melanins

Eric D Walberg* and Thomas J Volk. Department of Biology, 2036 Cowley Hall, University of Wisconsin-La Crosse, La Crosse, WI 54601.

Melanins are ubiquitous pigments whose unique structure, as random hydrophobic biopolymers with long-range delocalization, imparts an array of benefits. These include UV protection, cell wall strength and recalcitrance, desiccation tolerance, penetration of hosts' more potent physical defenses, and others. From the halophilic to the pathogenic, from the arctic to hot deserts, from bare rock to soaked soils, melanins are useful in many habitats and situations. Fungi reaping benefits from melanin granules or layers in their cell walls exist in abundance in most if not all major fungal groups. The zygospores of zygomycetes like Rhizopus, the appressoria of plant pathogenic ascomycetes like Magnaporthe, the spores of many fruiting basidiomycetes such as Coprinus, the macrospores of endomycorrhizal Glomeromycota such as Glomus, the resting structures of the rumen-inhabiting Neocallimastix, and, of course, the entirety of the form-famly Dematiaceae are all examples of major groups and genera making use of melanins. Melanins can be produced with a variety of substrates and methods, from common DHN (1,8-dihydroxynaphthalene) produced in many ascomycetes via pentaketide synthesis, to many suitable exogenous substrates used by Cryptocuccus neoformans with its promiscuous laccase. With such a diversity of fungi making use of them in a variety of ways, melanins deserve more attention in the mycological community and classroom. In this educational poster, we will present and examine some fungal melanins, some of their many uses, and some organisms that use them.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P84: Viability and efficacy of probiotic yeast supplements from U.S. retail

V H Tournas1*, Naomi Santillan2, and Nicholas S Niazi3. 1CFSAN/FDA, 5100 Paint Branch Parkway, College Park, MD, USA, 2HACU/University of Texas, Rio Grande Valley, TX, USA, 3JIFSAN/University of Maryland, College Park, MD, USA.

Yeast (Saccharomyces boulardii) probiotic preparations from three U.S. companies (A, B and C) were tested for viability and efficacy. A total of 33 samples were analyzed for levels of live organisms per gram and per capsule using the dilution plating method. All samples were tested before their expiration dates. Results showed that overall S. boulardii levels ranged between 2.6 x 109 to 4.0 x 1010 colony forming units per gram (cfu/g) or 1.4 x 109-8.0 x 109 cfu/capsule. Probiotic supplements from company A had S. boulardii levels similar to/or higher than those on the label, while all samples from company B had significantly higher counts than the ones claimed on the label. Most samples from company C contained significantly lower yeast counts than the ones claimed on their respective labels. Variation in viable yeast levels existed among companies, among lots from the same company as well as within lots.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P85: Mycological Research in Panama: A high-impact field experience for students with variable levels of training

Alma E Rodriguez Estrada1*, Michael L Draney1, and Vicki Medland2. 1University of Wisconsin Green-Bay. Department of Natural and Applied Sciences ES-317. 2420 Nicolet Dr. Green Bay, WI 54311, 2University of Wisconsin Green-Bay. Cofrin Center for Biodiversity. Mary Ann Cofrin Hall 218E. 2420 Nicolet Dr. Green Bay, WI 54311.

Research experiences for undergraduate students have been recently recognized as a high-impact educational practice to accomplish excellence in any academic field. The University of Wisconsin-Green Bay encourages research experiences and international courses where undergraduate students have the chance to fully participate in formal scientific work while experiencing life in a different culture. The upper level course "Research Experience in Panama" is offered every year in January. During this course, faculty members and students (biology and environmental science majors) travel to Panama to perform research for two weeks. For four years, research themes related to mammals, birds, fish, spiders, and coral reef and seagrass benthic invertebrates. In 2012, mycological research was implemented for the first time. This work utilizes a protocol that allows students with different levels of mycological training (undergraduate and master-level students, inexperienced and students who took an upper level mycology course) to help with ecological macromyetes data collection. This protocol also allows to rapidly assess the diversity of ground-accessible macromycetes using standardized, repeatable and randomized samples. Data were collected to study the ecology of ectomycorrhizal and saprotrophic macrofungi at a variety of sites near Bocas del Toro and Gamboa field stations (Smithsonian Tropical Research Institute). Specifically, this research seeks to study the effects of altitude and vegetation on macromycetes richness and abundance across a range of Panamanian rainforest (lowland, mid and high elevation). Parallel transects of 250 m2 (50 x 5 m) were delimited within 0.25 hectare sites where macromycetes were collected, described and counted. The sites sampled in 2012 will be sampled in subsequent years, and different students will participate. Ecological research on macromycetes is a high-impact practice that strengths students' understanding of science and elevates students' awareness of the importance of fungi in the ecosystems.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P86: Trametes versicolor: A tremendously versatile fungus

Andrew M Hart* and Thomas J Volk. Department of Biology, University of Wisconsin, La Crosse, WI 54601, USA.

Since microbes are generally seen in a negative public light, it is refreshing to take a look at one that is not only beneficial, but has the potential to serve multiple human interests. Trametes versicolor is a white rot polypore commonly referred to as the "turkey tail" mushroom. It is named for and characterized by the many concentric zones of color on the fruiting body. It is important to note that due to its prevalence in the northern forests, the turkey tail is an important saprophyte, but this might be the least useful quality of T. versicolor. The same versatile enzyme (laccase) that degrades lignin is key to several of T. versicolor's applications. Laccases are used in biopulping and the bioremediation of a variety of environmental contaminants. T. versicolor produces this valuable enzyme, and also gives us polysaccharide-K (PSK). Polysaccharide-K is anecdotally proven to function as an adjuvant (immunostimulatory). Polysaccharide-K is thought to enhance immune response to cancer cells. In addition to medical and environmental uses, T. versicolor is also used for its pigments. The pigments increase the wood's monetary value and in turn are prized by woodworkers and artists. The inoculated wood forms streaks of color, called spalting. T. versicolor pigments are also used to dye fabrics such as wool. The multi-colored velvety fruiting body is used to make art of all kinds. These are the uses we've found for the turkey tail so far. As more research is done we will see the common fungus even more. This poster is applicable for education in courses related to Mycology, Medicine, Bioremediation and Biodegradation, Environmental Toxicology, Ecology, Biochemistry, and Biology.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P87: A preliminary checklist of Agaricales from Reserva Biológica do Alto da Serra de Paranapiacaba, Santo André, SP, Brazil

Marina Capelari1*, Fernanda Karstedt1, Jadson JS Oliveira1, and Nelson Menolli, Jr.1,2 1Instituto de Botânica, Núcleo de Pesquisa em Micologia, Caixa Postal 68041, 04045-972 São Paulo, SP, Brazil, 2Instituto Federal de Educação, Ciência e Tecnologia de São Paulo, Campus São Paulo, CCT / Biologia, Rua Pedro Vicente, 625, 01109-010 São Paulo, SP, Brazil.

The Atlantic Forest is a long but discontinuous strip of tropical rainforest more than one hundred kilometers wide along the coast, from north to south Brazil. The "Reserva Biológica do Alto da Serra de Paranapiacaba" located at 23°46´00´´-23°47´10´´S and 46°18´20´´-46°20´40´´W, comprises 336 ha of this forest, with an annual rainfall and temperature of 3,300 mm and 14-15°C in the winter to 21-22°C in the summer, respectively. Altitude varies from 750 m to-891 m and the vegetation comprises high forests, low forests, scrub, dark scrub and grassland in different successional stages. The fungi were collected randomly across several trails in the forest, during the period of April/2006 to April/2010, with ca. of 367 collections. 140 collections represent 52 species belonging to 18 genera (Calliderma, Collybia, Crepidotus, Cyptotrama, Cystoderma, Gymnopus, Hypholoma, Inocephalus, Leptonia, Marasmius, Melanotus, Mycena, Nolanea, Pholiota, Pluteus, Pyrrhoglossum, Ripartitella and Stropharia). The remainder belong to, at least, 32 genera (Agaricus, Amanita, Camarophyllus, Collybia s.l., Conocybe, Coprinus, Crepidotus, Dictyopanus, Favolaschia, Filoboletus, Gerronema, Hygrocybe, Gloiocephala, Gymnopilus, Gymnopus s.l., Hypholoma, Lepiota, Leptonia, Leucocoprinus, Marasmiellus, Marasmius, Melanotus, Mycena, Pholiota, cf. Physocystidium, Pluteus, Psathyrella, Psilocybe, Rimbachia, Trogia, Tetrapyrgos and Tubaria). New taxa were recorded, some of them, already described (Calliderma fibulatum Karstedt & Capelari, Marasmius cystidioccultus J. S. Oliveira & Capelari, Marasmius plenicystidiosus J. S. Oliveira & Capelari), and some are in process of publication, especially of the genus Marasmius.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P88: Habitat diversity of macrofungi in the Indian part of Sundarban

Krishnendu Acharya1*, Arun K Dutta1, Prakash Pradhan1, and Anirban Roy2. 1Molecular and Applied Mycology and Plant Pathology Laboratory, Department of Botany, University of Calcutta, Kolkata –700 019, INDIA, 2West Bengal Biodiversity Board, Paribesh Bhawan, Salt Lake City, Kolkata – 700 098, INDIA.

The Sundarban Biosphere Reserve covers an area of approximate 25,500 sq km. However, only 9,630 sq km is in India, with the rest located in Bangladesh. The area in India is demarcated by the Hoogly River in the west, the Bay-of-Bengal in the south, and the Harinbhanga and Raimangal Rivers in the east and the Dampier-Hodges line in the north. The wide range of phyto-topographical features, presence of spatial and temporal variability in hydrological regimes and the diverse substrata has contributed to the high biodiversity of this region. Mangrove forest dominates in the core area and some margins of the deltic island, where inhabited lands show typical coastal vegetation. Several anthropogenic activities, recurring natural calamities and recent incidences of cyclones (e.g., Aila) have added to the vulnerability of this biologically rich world heritage site. The climate is characterized by relatively high temperature and humidity (>80%) throughout the year, and well distributed rainfall during the monsoon season. Temperatures rise from a daily minima of 10ºC in the winter to a maximum of about 43ºC in March and may exceed 32ºC during the monsoon season. The average annual rainfall in the Indian part of the Sundarban region is 1,662 mm. During our survey (2010-2011), 77 quadrates were studied each measuring 20 × 20 meters. Macrofungi were collected, photographed, identified and were preserved with accession numbers. A total of 71 species of macrofungi belonging to 29 families, 53 genera were identified and their ecology was evaluated.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P89: Cystolepiota and Echinoderma: tropical spp. in México

Joaquín B Cifuentes* and Rosalva A Vázquez-Estup. Biología, Facultad de Ciencias, UNAM, México DF.

Cystolepiota as currently defined (Vellinga, 2007) contains between 10 and 55 taxa (Kirk et al., 2008; Vellinga, 2010). Echinoderma is considered either a synomyn of Lepiota (Kirk et al., op. cit., Vellinga, 2010) or a separate genus with up to 14 taxa (Bon, 1992; Knudsen et Versterholt, 2008). Phylogenetic analysis suggested a close relationship between these genera (Vellinga, 2003). Only the following species have been reported from Mexico (under different genera): C. acutesquamosa, C. cygnea and L. hemisclera (Herrera and Guzmán, 1972; Bandala et al., 1988; Cifuentes, 2008). Our ongoing revision of lepiotaceous fungi from Mexico has revealed some interesting tropical taxa in these genera. Here we describe and illustrate two taxa in Cystolepiota (one C. aff. petasiformis and a putative new taxon) and two putative new taxa in Echinoderma.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P90: One hundred and twelve years of Pluteus knowledge in Brazil: revision of the first collections studied by J. Rick and P. Hennings

Nelson Menolli, Jr.1,2* and Marina Capelari1. 1Instituto de Botânica, Núcleo de Pesquisa em Micologia, Caixa Postal 68041, 04045-972 São Paulo, SP, Brazil, 2Instituto Federal de Educação, Ciência e Tecnologia de São Paulo, Campus São Paulo, CCT / Biologia, Rua Pedro Vicente 625, 01109-010 São Paulo, SP, Brazil.

The genus Pluteus comprises ca. 300 species worldwide of which 70 have been mentioned from Brazil; however, it is believed that about 23 are certainly known. To solve some of the taxonomic problems, techniques of molecular biology can be used, but there is some limitations e.g. when studying old collections, and an accurate morphological study may be an alternative. Complementing the Brazilin knowledge of Pluteus, the first collections studied in the early 20th century were re-examined. At first, Hennings described P. scruposus, P. cervinus var. griseoviridis and P. termitum. Later, between 1907 and 1961, Rick recorded 21 more Pluteus taxa. The holotypes of P. scruposus and P. termitum deposited in Berlin were probably destroyed. Collections of nine taxa recorded by Rick (P. cervinus var. patricius, P. cristatulus Rick, P. eximius, P. fibrillosus Rick, P. melanodon, P. nanus var. podospileus, P. umbrosus) also were not found. All other collections are deposited at PACA and SP and despite the bad conservation, some micromorphological structures were recovered and the following considerations could be made correcting the European names attributed in the past by Rick and Hennings. P. brunneopictus: is probably P. tucumanus Singer; P. cervinus and P. cervinus var. griseoviridis: represent P. xylophillus (Speg.) Singer; P. exiguus: is probably P. jamaicensis Murrill; P. granulatus: is possibly P. glaucotinctus Horak; P. hispidulus: is probably P. yungensis Singer; P. leptonia Rick: is a species of Entoloma s.s.; P. nanus: is possibly P. pulverulentus Murrill; P. pellitus: is probably P. petasatus; P. phlebophorus: represents two collections which probably are P. pulverulentus and P. tucumanus; P. sensitivus Rick: is probably P. albostipitatus; P. straminellus Rick: is a nomen dubium; P. velatus Rick: is a nomen dubium; P. wehlianus: is a species of Bolbitiaceae or Strophariaceae.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P91: New species of agarics from the Páramo region in Colombia

Timothy J Baroni1*, Ana Esperanza Franco-Molano2, Marcelo Betancur2, and Tatiana Sanjuan2. 1Department of Biological Sciences, P. O. Box 2000, State University of New York - College at Cortland, Cortland, NY 13045, USA, 2Laboratorio de Taxonomía y Ecología de Hongos, Instituto de Biología, Universidad de Antioquia, A.A. 1226, Medellín, Colombia.

In May of 2011 a one-day field excursion was made to the Páramo ecosystem (Luteyn, 1999) in the department of Tolima, Colombia, as part of a longer research excursion to the cloud forest mountainous regions of south central Colombia. Fifteen collections of mostly agarics were made in the Páramo, several growing directly on decaying leaves of standing Espeletia. Two of these collections are clearly new species, one a Melanotus, the other a Hypsizygus. These new taxa will be illustrated and the other collections found on that excursion will also be displayed and discussed. This isolated area with its distinctive vegetation clearly deserves further exploration over longer time periods as it appears to be under explored.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P92: Lepiotaceous fungi (Agaricaceae, Basidiomycota) in Northeastern North America

Alfredo Justo. Clark University. Biology Department. 950 Main St, 01610, Worcester, MA.

Collections of Lepiotaceous fungi (Lepiota and allied genera in the Agaricaceae) made in the Northeastern U.S.A between 2009-2011 were studied using morphological and molecular (nrITS) data. Two taxa are currently under revision and probably represent undescribed species: (i) Lepiota novae-angliae, nom. prov., recorded from Massachusetts and New Hampshire closely resembles L. clypeolaria in morphology but it's ITS sequences do not match those of L. clypeolaria or any of the species of sect. Lepiota currently available in GenBank; (ii) Leucoagaricus allochthonus, nom. prov., was recorded in an indoors (greenhouse-like) garden in central Massachusetts and is probably a tropical alien species. Microscopically, it is characterized by the presence of pleurocystidia, a feature only shared by 3-4 other species in the very diverse Leucoagaricus/Leucocoprinus lineage (> 100 described species). Molecular data confirmed the presence of widespread cosmopolitan species in our area (e.g. Lepiota cristata, Leucoagaricus leucothites) but also revealed molecular differences between isolates of northeastern lepiotaceous fungi (e.g. Leucoagaricus rubrotinctus, Lepiota felina) and GenBank sequences deposited under the same name but with a different geographic origin.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P93: Resolving the generic boundaries within the Rhodocybe/Clitopilus clade (Agaricomycetes, Basidiomycota)

Kerri L Kluting1*, Sarah E Bergemann1, and Timothy J Baroni2. 1Middle Tennessee State University, Biology Department, PO Box 60, Murfreesboro TN 37132, 2State University of New York, College at Cortland, Department of Biological Sciences, PO Box 2000, Cortland NY 13045.

The Entolomataceae (Agaricomycetes, Basidiomycota) is a highly diverse assemblage of euagarics with well over 1000 taxa worldwide. These macrofungi are recognized as a cohesive unit by their pink or flesh brown basidiospores that are angular in polar view, have some form of a ridged or pustulate ornamentation and the walls of the spores are evenly cyanophilic. Recent published phylogenetic analyses support the separation of these taxa into two clades, typically referred to as the 'Entoloma' and 'Rhodocybe/Clitopilus' clades. The Rhodocybe/Clitopilus clade is the focus of our investigation, as a recent publication suggests that the genus Rhodocybe (pustulate spores) should be subsumed under the genus Clitopilus (longitudinally ridged spores) in order to form a monophyletic clade. Our goal is to conduct a systematic revision of the Rhodocybe/Clitopilus clade, focusing on delimitation of generic boundaries using a phylogenetic framework. Initial efforts have focused on obtaining sequences of mitochondrial and low-copy nuclear loci [mitochondrial small subunit ribosomal RNA (mtSSU) and ATPase subunit 6 (ATP6), the nuclear RNA polymerase second largest subunit (RPB2) and the translation elongation factor alpha gene (TEF1-α)] to define the generic boundaries. To date, we have sequenced and performed a phylogenetic analysis utilizing 86 collections of Clitopilus and Rhodocybe. The results of our combined analysis of the four genes provide significant support for the delineation of the following clades within Rhodocybe: i) a clade containing Clitopilopsis hirneola (formerly as Rhodocybe hirneola); ii) a fallax/mundula/popinalis clade nested between Clitopilus and Clitopilopsis; iii) a Rhodocybe s.s. clade with several Rhodocybe taxa, including the type for the genus, R. caelata and; iv) a clade containing R. nitellina. A discussion of possible scenarios to explain evolution in the Rhodocybe/Clitopilus clade will be presented.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P94: Lentinus sensu stricto in Southeast Asia

Jaya Seelan Sathiya Seelan1,2*, Alfredo Justo Justo1, and David Hibbett Hibbett1. 1Biology Department, Clark University, Worcester, MA 01610, 2Institute for Tropical Biology and Conservation, Universiti Malaysia Sabah, Jalan UMS 88400, Kota Kinabalu, Sabah, Malaysia.

Lentinus Fr. (Basidiomycota, Polyporales) is a genus of saprotrophic, normally wood-decaying, mushroom-forming fungi characterized by the decurrent lamellae, dimitic tissues in the basidiome, and hyaline, ellipsoid to cylindric spores. The genus seems to be particularly diverse in tropical areas. The generic limits of Lentinus and the boundaries between the species are currently under discussion. We will present the results from our study on the biodiversity and systematics of Lentinus in Southeast Asia (Malaysia, Indonesia, Papua New Guinea and Thailand) using morphological and molecular (nLSU, nrITS) characters and discuss the implications for the species-level and generic taxonomy of Lentinus.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P95: Geographic distribution of Fomes fasciatus and F. fomentarius in the United States

Meghan A McCormick*, Marc A Cubeta, and Larry F Grand. North Carolina State University, Department of Plant Pathology, Box 7567, Raleigh, NC 27695.

Fomes fasciatus and F. fomentarius can cause a white heart rot in multiple species of trees and play important roles in forest ecology and management. F. fasciatus is distributed across the Southeastern U.S., throughout Central America and into South America., while F. fomentarius is distributed throughout the boreal forests of the Northern Hemisphere. North Carolina is a unique geographic transition area for these species as it represents the northern limit for F. fasciatus and the southern limit for F. fomentarius. In this study, U.S. distribution maps (by county) of F. fasciatus and F. fomentarius were developed based on records from 26 mycological herbaria, publications, and collections made for this study. The geographic distribution for both species was expanded to include six counties in four states not included in previous publications. Both species are associated with a diverse range of trees, which includes hardwood species from multiple genera. In the US, F. fasciatus has been reported predominantly on species of oak (Quercus) and hickory (Carya), while F. fomentarius typically decays species of birch (Betula), beech (Fagus), and maple (Acer). The distribution of host species in many cases extends beyond the known limits of the distribution of F. fomentarius and F. fasciatus indicating that other delimiting factors contribute to the distribution and occurrence of these fungi.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P96: Species delimitation in Trametes (Polyporales, Basidiomycota): a comparison of ITS, TEF1, RPB1 and RPB2 phylogenies

Alexis L Carlson*, Alfredo Justo, and David S Hibbett. Clark University. Biology Department. 950 Main St, 01610, Worcester (MA).

Trametes is a cosmopolitan genus of white rot polypores, including the "turkey tail" fungus, T. versicolor. Although Trametes is one of the most familiar genera of polypores, its species-level taxonomy is unsettled. The ITS region is the most commonly used molecular marker for species delimitation in fungi and, despite some problems and limitations, it generally offers a good level of resolution. During our phylogenetic study of the genus Trametes we observed a low level of molecular variation in the ITS that resulted in poorly resolved phylogenies and unclear species boundaries, especially in the Trametes versicolor species group (T. versicolor, T. ochracea, T. pubescens, T. ectypa). In this study we will evaluate the performance of three protein-coding genes (TEF1, RPB1, RPB2) for species delimitation and phylogenetic reconstruction in Trametes. Preliminary results suggest that all three protein-coding genes outperform ITS for separating species in the T. versicolor complex

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P97: Cryptomarasmius gen. nov. is proposed in the Physalacriaceae to accommodate members of Marasmius section Hygrometrici

Thomas S Jenkinson1,2*, Brian A Perry3, Rainier E Schaefer1, and Dennis E Desjardin1. 1Department of Biology, San Francisco State University, 1600 Holloway Ave., San Francisco, CA 94132, 2Department of Ecology and Evolutionary Biology, University of Michigan, 830 North Universty Ave., Ann Arbor, MI 48109, 3Department of Biology, University of Hawaii, Hilo, 200 West Kawili St., Hilo, HI 96720.

Phylogenetic placement of the infrageneric section Hygrometrici (genus Marasmius sensu stricto) in prior molecular phylogenetic studies have been unresolved and problematical. Molecular analyses based on newly generated ribosomal nuc-LSU and 5.8S sequences resolve members of section Hygrometrici to the family Physalacriaceae, within the Agaricales. The new genus Cryptomarasmius is proposed in the Physalacriaceae to accommodate members of Marasmius section Hygrometrici. Taxonomic transfers of fourteen species within section Hygrometrici are recommended to reflect phylogenetic affinity.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P98: Stuff in Duff: a metagenomic study of fungi in kauri forests

Mahajabeen Padamsee1*, Stanley E Bellgard2, Ian Dickie3, Scott Fraser4, Duckchul Park1, Bryan Stevenson4, and Bevan Weir1. 1Biosystematics Team, Landcare Research, Auckland, New Zealand, 2Biodiversity and Conservation Team, Landcare Research, Auckland, New Zealand, 3Ecosystem Processes, Landcare Research, Lincoln, New Zealand, 4Soils and Landscape Responses, Landcare Research, Hamilton, New Zealand.

Kauri (Agathis australis, Araucariaceae) is restricted in distribution to the northern tip of the North Island of New Zealand. Living to 2,500 years or more and having trunks to 7 m diam., kauri is a keystone species. Kauri was extensively logged and now less than 1% remains of undisturbed old-growth forests. Since the 1970s these trees have also been under threat from the exotic invasive, Phytophthora taxon Agathis (PTA) that causes kauri dieback. We are interested in understanding how the fungal community associated with the large amount of kauri duff (i.e., leaf litter) will be affected after infection of the tree by this exotic root pathogen. However, there has been no thorough study of the fungi associated with kauri and as a result we have no baseline data of the composition of a "healthy" fungal community in kauri leaf litter. We used pyrosequencing to study fungal biodiversity and assess changes in fungal community composition after invasion by PTA. The sampling scheme was optimized and several methods of analyzing the data were tested. A sequence database of known fungi was created with which to compare the metagenomic data.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P99: Resolving the phylogenetic relationship between Armillaria and the newly discovered genus Guyanagaster

Rachel A Koch1*, Terry W Henkel2, and M Catherine Aime1. 1Department of Plant Pathology and Crop Physiology, Louisiana State University Agricultural Center, Baton Rouge, Louisiana 70803 USA, 2Department of Biological Sciences, Humboldt State University, Arcata, California 95521 USA.

Recent mycological work in the remote Pakaraima Mountains of Guyana uncovered Guyanagaster necrorhiza, a new gasteromycete genus and species in the Physalacriaceae (Agaricales). The original study showed that Guyanagaster is most closely related to the agaricoid genus Armillaria and has a similar root parasitism/wood decay habit. While Armillaria species exhibit typical lamellate mushroom morphologies Guyanagaster basidiomata are entirely sequestrate with a thick, hard peridium, tough, decay-resistant gleba with highly modified hymenium and heavily ornamented basidiospores, and a short stipe attached directly to root substrata. Molecular phylogenetic analyses using different gene regions and varying numbers of taxa were, however, not in complete agreement as to the exact relationship between Guyanagaster and Armillaria; some analyses suggested that Guyanagaster is sister to Armillaria, whereas others indicated that Guyanagaster is nested within Armillaria. To better understand the phylogenetic relationships of Guyanagaster and Armillaria within the Physalacriaceae as well as implications for evolution of basidioma forms, we built an expanded three-gene supermatrix (ITS, LSU and EF1α) from an increased number of Armillaria species, Guyanagaster, and other exemplar Physalacriaceae taxa. A maximum likelihood analysis of these data suggested that Guyanagaster may be derived from within Armillaria, a result consistent with demonstrated origins of sequestrate taxa within other lamellate lineages. In light of these results we feel that retaining generic status for Guyanagaster is warranted due to its extreme morphological divergence from Armillaria; such an approach will, however, render Armillaria sensu lato paraphyletic. We also present a new species of Armillaria that occurs sympatrically with Guyanagaster and is the first record of an Armillaria species in Guyana.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P100: Fungal diversity associated with oaks in Korea

Young Woon Lim1*, Hyun Lee1, Changmu Kim2, and Jin Sung Lee2. 1School of Biological Science, Seoul National University, Seoul, 151-747, Korea, 2National Institute of Biological Resources (NIBR), Incheon, 404-708, Korea.

Eleven species of oak trees are common in Korean forests in association with evergreen pines. Although they have great economic value, oak trees have received little attention. With the goal to manage oak forests effectively, we investigated the diversity of decay-causing fungi and ectomycorrhizal (EM) fungi from the eight species of oak trees. Through examination of the specimens deposited in the Seoul National University Fungus Collection (SFC) and the National Institute of Biological Resources (NIBR) of the Ministry of Environment, a total of 523 decay fungi (104 species) associated with oak trees were identified and listed. No host specificity of decay fungi was evident. However, many decay fungi preferentially attacked specific sites within oak trees. To investigate the diversity of the ectomycorrhizal fungi, cloning and pyrosequencing were carried out. Through the sequence analysis of 243 clones, 67 sequences were identified as the plant and 176 sequences (62 phylotypes) were the fungi (Basidiomycota-142 (43), Ascomycota-34 (19)). Major phylotypes were ecotomycorrhizal fungi such as Russula and Lactarius, and the others were litter/decay rotters and dark septate endophytes. A total of 17,694 sequence reads were obtained from pyrosequencing, where 9,951 were fungi and 5,597 were plant. Compared with RFLP and cloning, the pyrosequencing approach yielded more diverse results.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P101: Overview of research activities of the FunWood and FuPerS project consortia of the German Biodiversity Exploratories: fungal and bacterial diversity and wood and litter decay

Dirk Krüger1*, Björn Hoppe1, Witoon Purahong1, Danuta Kapturska1, Kristin Baber2, Marek Pecyna3, Tobias Arnstadt3, Renate Rudloff1, Harald Kellner3, Peter Otto2, Tiemo Kahl4, François Buscot1, Jürgen Bauhus4, and Martin Hofrichter3. 1UFZ - Helmholtz Centre for Environmental Research, Department of Soil Ecology, Th.-Lieser-Str. 4, D-06120 Halle/Saale, Germany, 2University of Leipzig, Institute of Biology I, Herbarium Universitatis Lipsiensis (LZ), Johannisallee 21-23, D-04103 Leipzig, Germany, 3International Graduate School Zittau – IHI, Markt 23, D-02763 Zittau, Germany, 4University of Freiburg, Institute of Silviculture, Tennenbacherstr. 4, D-79085 Freiburg i. Brsg., Germany.

In forests ecosystems, coarse woody debris (CWD, a.k.a. dead wood) and litter are dominant habitats rich in lignin and cellulose. Their degradation, mainly by Fungi, is an important part of nutrient cycling. Intensive use of forests for timber production increases loss of CWD, thus may affect the diversity of organisms sharing this habitat. The FunWood project of the DFG (German Science Foundation) Biodiversity Exploratories investigates the postulated effect of forest management intensity on the diversity of wood-decaying fungi and dead wood decomposition. In the first project phase the fungal diversity was approached both by molecular biological methods (fingerprinting and sequencing of clone libraries) and surveying fruit bodies. In addition, preliminary investigations on bacterial diversity, including nitrogen fixation genes (nifH), were performed. Potentially new bacteria were identified by nifH. The follow-up, second phase of this project (started 2011) scales up the study on wood decay fungi to encompass 150 plots in all three exploratory locations (Schwäbische Alb, Hainich-Dün, Schorfheide-Chorin). Thirteen species of trees whose logs were placed there as the BeLongDead experiment are now included. A two-year experiment of adding nitrogen to dead wood was also initiated. We discuss some results of phase 1 and some planned activities in phase 2. In addition, some current aspects of a related project, FuPerS, studying leaf litter degradation, are discussed.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P102: Fungal communities of Populus - comparisons between culture-based and culture independent approaches

Khalid Hameed1*, Gregory Bonito1, Michael Gajdeczka1, Christopher Schadt2, and Rytas Vilgalys1. 1Biology Department, Duke University, Durham NC 27708, 2Oak Ridge National Laboratory, Department of Energy, Oak Ridge TN 37831.

Populus is an economically important genus of woodland and riparian trees that forms both arbuscular and ectomycorrhizas. Populus also harbors a wide-diversity of root endophytes but this diversity is still largely uncharacterized. To address this field and greenhouse studies were conducted on different Populus species and genotypes. Fungal community diversity in roots was determined with culture-independent 454 pyrosequencing of phylogenetically informative fragments of genomic rDNA (ITS, LSU, SSU). Companion culture-based studies of Populus root associates were carried out to: 1) describe the culturable diversity of Populus root associates; 2) determine discrepancies between culture-based and culture-independent methods; 3) obtain a culture library of fungal root associates of Populus for future experiments and genome sequencing. Using sterile technique and selective isolation medias, we isolated >300 cultures from individual surface sterilized root tips of Populus deltoides collected from our field site and greenhouse experiments. We also isolated fungi from roots of Pinus taeda (loblolly pine) and Quercus alba (white oak) for comparison. Pure cultures were sequenced at the ITS and LSU. DNA sequences were identified by comparing them to accessions in the NCBI nucleotide database and to sequences assembled from pyrosequencing data. We estimate our cultures represent ~65 OTUs based on 97% similarity of the ITS. Despite using benomyl in our media to select for basidiomycetes, the majority of OTUs from sequenced isolates were ascomycetes. We obtained cultures from a number of the fungi that were among the most abundant sequence type in our 454 datasets (e.g., Atractiellales spp., Ilionectria spp., Mortieriella spp, and Thelephoraceae sp.). Further, fruitbodies of Laccaria were found under a number of trap-plants and this species has been obtained in pure culture. These isolates will be valuable for further studies on Populus-microbial relations.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P103: Complete genome sequence of Pleurotus eryngii using the next generation sequencing

Jae San Ryu1*, Song Hee Lee2, Kyung Hee Kim1, Hee Jeong Je1, Min Keun Kim1, Hyun sook Lee2, and Su Myung Chae1. 1Eco-friendliness Research Department, Gyeongsangnam-do Agricultural Research and Extension Services, Jinju 660-360, Republic of Korea, 2Departments of Microbiology, Gyeongsang National University, Jinju, 660-900, Republic of Korea.

Pleurotus eryngii (King oyster mushroom) is an edible mushroom with potential health benefits that thrives on a parasitic host in the steppe area. P. eryngii is a basidiomycete with a tetrapolar incompatibility system comprising two mating type loci, A and B. Two monokaryons P5 and P6 from P. eryngii KNR 2312 strain by a de-dikaryotization were analyzed by CHEF, revealing 10 chromosomal bands with a size range from 3.0 to 7.1 Mbp and an estimated total genome size of 43.8 Mbp. GS-FLX and HiSeqTM Sequencing analysis of P5 as a Next Generation Sequencing (NGS) strategy was performed to clarify its genetic composition. Shotgun and mate paired libraries were constructed for NGS. The read count was 650,812 and 433,267,606 total bases for the shotgun library. Average length of read was over 665 bases. For the mate paired library, read count was 122,368, total bases was 47,889,870, and average of length was 391.36. Scaffolding was performed to assemble contigs and mate paired data. The number of scaffolds and bases were 1,506 and 43,991,976, respectively. The scaffold accounted for 100% of the genome size estimated by karyotyping. To estimate function, sequences were compared with those in the NCBI blastx database. Over 1,000 genes were aligned with the sequences within the contigs. Functional genomics will be required to elucidate unknown genes. Results of this genome project will soon allow sequence based breeding like MAS (marker assisted selection).

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P104: Mobyle SNAP Workbench: a phylogenetic and genomic analysis web portal

James T Monacell and Ignazio Carbone*. Department of Plant Pathology, North Carolina State University, Raleigh, NC 27606.

The SNAP Workbench toolkit is a stand-alone software application that integrates a wide array of bioinformatics tools for phylogenetic and population genetic analyses. We have developed a web-portal frontend, using the Mobyle portal framework, which runs all of the programs available in the stand-alone SNAP Workbench on a high-performance Linux cluster. Additionally, we have expanded the selection of programs to over fifty tools, including 1) a suite of genome assembly and analysis tools such as Fastx-Toolkit, Quake, Burrows-Wheeler Alignment Tool, Velvet, SOAPdenovo, GeneMark-ES, and Augustus, 2) large-scale phylogenetic methods as implemented in RAxML, and 3) metagenomic analysis tools such as Mothur and Esprit. The Mobyle SNAP Workbench web-portal allows researchers to seamlessly execute and manage otherwise complex command-line programs with multiple input files and parameters, as exemplified in the coalescent approaches implemented in Genetree, MDIV, IM, IMa, MIGRATE, and LAMARC; these tools are parameter-rich and therefore computationally intensive, often requiring several days or weeks to run. Our optimization includes 1) selecting appropriate number of compute nodes and machine architecture for MPI programs and 2) parallelization of computational tasks to execute on multiple machines when possible (e.g. Genetree). A unique feature of the portal is the implementation of workflows that link together several programs sequentially. This greatly facilitates exploratory analyses for the beginner but also allows for efficient use of cluster resources. The possibility of making the portal accessible to the public, as a web service on Amazon EC2 or at NC State, will be discussed.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P105: Open Tree of Life: Community driven synthesis of the Tree of Life

J Gordon Burleigh1, Keith Crandall2, Karen Cranston3, Karl Gude4, David S Hibbett5*, Mark Holder6, Laura A Katz7, Richard H Ree7, Stephen A Smith7, Douglas E Soltis7, and Tiffani Williams7. 1Department of Biology, University of Florida, Gainesville, FL 32611, 2Department of Biology, Brigham Young University, Provo, UT 84602, 3NESCENT, Durham, NC 27705, 4College of Communication, Arts and Sciences, Michigan State University, E. Lansing MI 45622, 5Biology Department, Clark University, Worcester MA 01610, 6Department of Ecology and Evolutionary Biology, University of Kansas, Lawrence KS 66045, 7and elsewhere..

Reconstructing the phylogeny of all species has been a grand challenge ever since Darwin. The scope of the problem is immense: current estimates of extant biodiversity range from 1.8 million to 8.7 million species, a large fraction of which are Fungi. Much progress has recently been made in resolving the tree, and systematists continue to generate new phylogenetic knowledge at all depths of ancestry. Nevertheless, despite 150 years of effort, 55 AToL projects, and numerous other funded projects, we lack a comprehensive tree of life. Synthesis is currently inhibited by limits of available data, analytical power, and informatics infrastructure. Perhaps more importantly, it is also limited by a lack of compelling means and incentives for community participation. A comprehensive synthesis would yield great benefits across the life sciences, especially if it were self-sustaining, community-driven, and continually updated. We describe a recently funded project named "Open Tree of Life" that aims to establish a community-driven, continually updated estimate of the entire tree, and develop new software tools and new methods for merging and sharing data. Open Tree of Life will: 1) within one year, compile the first comprehensive draft tree of life by synthesizing existing phylogenetic and taxonomic knowledge; 2) enable the community to improve, annotate, and expand this initial tree; 3) initiate a cultural transformation in systematics towards pervasive and ingrained practices of data sharing; and 4) develop novel methods for synthetic tree reconstruction. By engaging the systematics community, including mycologists, our overarching goal is to cultivate ongoing synthesis on a large scale, in a manner that will transform current cultural norms in the field.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P106: Contributions of ectomycorrhizal fungi to organic matter formation and degradation in response to chronic nitrogen deposition

Jesse J Sadowsky*, Linda TA van Diepen, and Serita D Frey. University of New Hampshire, 114 James Hall, Durham, NH 03824.

Nitrogen deposition promotes carbon storage in temperate forests by altering organic matter decomposition processes and decomposer communities. Ectomycorrhizal (ECM) fungi participate in soil organic matter dynamics by producing abundant mycelium and actively degrading labile and recalcitrant organic matter, but have not been considered explicitly for their role in organic matter contribution and decomposition in simulated long-term nitrogen enrichment studies. Plots within a temperate mixed-hardwood stand at the Harvard Forest Long-Term Ecological Research site in Petersham, MA, USA have been amended with 50 or 150 kg N ha-1 yr-1 since 1988 to simulate projected future N deposition levels or N saturation, respectively. We determined how chronic N deposition affects annual organic matter input by ECM fungi by collecting hyphae in root-excluded sand bags. We assessed potential organic matter decomposition by ECM fungal communities by measuring activity of five C, N, or P hydrolases and two oxidases on the mantle of ECM root tips and of foraging extraradical mycelium collected in sand bags. Fungal symbionts were identified by DNA sequencing. ECM hyphal production was reduced at the highest nitrogen deposition rate. Nitrogen deposition increased cellulolytic and chitinolytic activity of ECM root tips in the organic soil horizon and decreased aminopeptidase activity in the mineral soil horizon. This generally agrees with earlier observations in bulk soil at this and other sites receiving multiple years of nitrogen enrichment. In root-excluded sand, chitinase activity increased by three-fold at both rates of added nitrogen, while laccase activity declined by one-third and three-fourths at low and high nitrogen rates. ECM fungal species richness was least at the highest nitrogen deposition rate. Further efforts towards determining the fate of organic matter pools contributed and decomposed by ECM fungi communities will help to clarify their role in soil organic matter dynamics under elevated nitrogen deposition and other anthropogenic disturbances.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P107: Estimating fungal contributions to nitrous oxide (N2O) emissions from nitrogen fertilized grasslands using tunable diode laser absorption spectroscopy.

Sara R Lopez1*, Robert L Sinsabaugh1, David T Hanson1, and Andrea Porras-Alfaro2. 1Department of Biology, University of New Mexico, Albuquerque, NM, 87131, 2Department of Biological Sciences, Western Illinois University, Macomb, IL, 61455.

The use of nitrogen (N) fertilizers has created international concern as denitrification of fertilizer-derived nitrate could result in higher rates of N2O release to the atmosphere. The discovery that many eukaryotic fungi can transform N through nitrification and denitrification pathways is significant from both biochemical and ecological perspectives, because fungi dominate microbial metabolism in many terrestrial ecosystems. It is known that roots from semi-arid grasslands are colonized by a diverse fungal community, and suppression of such fungal biomass can cause a substantial reduction of N2O production. N2O production by six soil fungal species was estimated by analyzing gas samples from pure cultures isolated from blue-grama rhizosphere soils at the Sevilleta Long-Term Ecological Research site in Central New Mexico. Cultures were maintained in liquid medium containing nitrate as N source and incubated at 25°C for 10 days prior to sealing the flasks to limit aeration. Gas samples obtained after 24, 48 and 72 hours of incubation were injected into a Tunable Diode Laser (Campbell Scientific). CO2 production was recorded to normalize N2O generation by fungal respiration. Preliminary results indicate that there is a high variability in N2O production among species but, overall, N2O production tends to increase with time of incubation. After 72 hours of incubation, the highest levels of N2O and CO2 were produced by Gibberella avenacea (4.63x10-6 μmol 100 ml-1 mg dry mass-1 and 0.3 μmol 100 ml-1 mg dry mass-1 respectively) and an unidentified group (Genbank AJ875391; 4.07x10-4 μmol 100 ml-1 mg dry mass-1 and 4.95 μmol 100 ml-1 mg dry mass-1 respectively.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P108: Lead isotopic composition of macrofungi: possible applications in fungal ecology

Jan Borovička1,2*, Martin Mihaljevič3, and Milan Gryndler4. 1Academy of Sciences of the Czech Republic, Nuclear Physics Institute, v.v.i., Department of Nuclear Spectroscopy, Řež 130, CZ-250 68 Řež near Prague, Czech Republic, 2Academy of Sciences of the Czech Republic, Institute of Geology, v.v.i., Laboratory of Environmental Geology and Geochemistry, Rozvojová 269, CZ-165 00 Prague 6, Czech Republic, 3Charles University, Faculty of Science, Institute of Geochemistry, Mineralogy and Mineral Resources, Albertov 6, CZ-12843 Prague 2, Czech Republic, 4Academy of Sciences of the Czech Republic, Institute of Microbiology, v.v.i., Laboratory of Fungal Biology, Vídeňská 1083, CZ-142 20 Prague 4, Czech Republic.

Macrofungi are able to effectively take up various trace elements, including toxic heavy metals, translocate them through mycelia and deposit them in fruiting bodies. This ability is species-specific; certain macrofungal species are effective accumulators of such metals as cadmium, zinc, mercury, silver, gold and vanadium. Despite growing at sites with an arsenic/silver soil concentration at "background" levels, several macrofungal species have been reported to hyperaccumulate arsenic or silver. The mechanisms of uptake and transport of the accumulated elements are unclear. However, a lead isotopic analysis might represent an efficient tool for tracing the origin of elements in soil profile. Lead is a non-essential toxic metal whose biogeochemical cycle has been affected to a great degree by man and lead isotopes have been thus introduced as "fingerprints" of environmental pollution. Each source of lead can have distinct or sometimes overlapping isotopic ratio ranges and the isotopic composition of lead in soils reflects a mixing of these sources. At sites with contrasting 206Pb/207Pb and 208Pb/206Pb ratios in particular soil horizons, the isotopic composition of macrofungal fruiting bodies might be a useful tool for tracing lead origin. Our preliminary data from a case study in Agaricus bernardii have supported this hypothesis. Furthermore, the isotopic composition of lead in saprotrophic macrofungi (Agaricus campestris, A. bernardii, A. xanthodermus and Leucoagaricus leucothites) of various origins has reflected more or less the expected pollution sources.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P109: Diversity and heavy metal content of macrofungi found on serpentine andgranite outcrops, Deer Isles, Maine, USA

Katherine E Jumper1*, Nishanta Rajakaruna2, and David Porter3. 1122 Cottage Street Bar Harbor, ME 04609, 2College of the Atlantic 105 Eden Street Bar Harbor, ME 04609, 3Department of Plant Biology University of Georgia Athens, GA 30602.

Fungal diversity in relation to edaphic conditions has rarely been studied. We are conducting a year-long comparative survey of macrofungi on serpentine and granite outcrops to determine whether there are differences in fungal biodiversity based on edaphic factors, and whether fungi tissue differ in heavy metal content based on the substrate upon which they are found. Previous studies we have conducted document differences in diversity of lichens, bryophytes, and vascular plants at these two sites, including significant differences in heavy metal accumulation in vascular plant species found at both sites. Serpentine soils contain high amounts of heavy metals and low amounts of essential nutrients and are a challenging environment for plant growth. Serpentine outcrops are known to harbor many rare and endemic plants, but very little is known about the ecology of macrofungi in serpentine habitats. We predict that we will find differences in species diversity between the two outcrops, reflecting differing abilities in the fungal species to adapt to the contrasting soil conditions. The poster presents our experimental design and findings from preliminary collections made during fall of 2011 and spring of 2012.

Poster Session 1
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2000-2100

P110: Airborne fungal spore dispersal and trait diversity in coastal mixed-evergreen forests in California

Sharifa G Crandall* and Gregory S Gilbert. University of California, Santa Cruz, 1156 High Street, Santa Cruz, CA 95064.

Although some studies characterize fungal spore dispersal dynamics in tropical forests, spore dynamics in temperate forests is still largely unknown. Airborne spore abundance in tropical forests is stratified: there is a higher density of spores in the still, dark, and moist understory compared to the windy, light, and dry conditions of the forest canopy. This spatial stratification suggests that spore dispersal is influenced by abiotic factors and that species may be dispersal limited. Moreover, once spores are discharged, specific spore traits such as size or ornamentation may restrict or facilitate spore movement. This study examines the spatial dynamics of airborne spores in a temperate coastal mixed-evergreen forest in California. We ask: 1) is there is a difference in spore density under different site conditions (open vs. closed canopy)? 2) do fungal spore traits differ among sites and vertically from the forest floor into the canopy? 3) which spore traits are phylogenetically conserved across taxa? Spores from different fungal species were trapped in closed versus open canopy forest sites at the University of California, Santa Cruz Upper Campus Reserve during the wet season in 2012. Spore density and traits are characterized by imaging of spores under a light microscope and analyzed using the software analysis tool imageJ®. Traits measured include spore diameter, size, outer ornamentation, cell wall thickness, and presence/absence of melanin. Spore samples will be identified using next generation sequencing (NGS) methods. We report on the spatial distribution of airborne spores and suggest phylogenetically conserved traits across fungal communities.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P111: The insect and yeast communities of carnivorous pitcher plants

Leonora S Bittleston and Anne Pringle*. 16 Divinity Avenue Cambridge, MA 02138.

Carnivorous pitcher plants are models for food web dynamics. The Northern pitcher plant, Sarracenia purpurea, has modified leaves, or pitchers, which are sterile until they open. Soon after opening the pitchers are filled with rainwater, and accumulate a diverse community of organisms. The microbes in pitchers are still relatively unknown, although recent studies have shown that a keystone predator, the mosquito Wyeomyia smithii, controls bacterial diversity. Numerous yeasts have been found within the pitchers, and they are different from those present in the surrounding bog water. We examined how insects affected the diversity and abundance of yeasts in S. purpurea pitcher plants. Insect exclusion with gauze coverings successfully excluded W. smithii, and the abundance of yeasts was positively correlated with insect counts. The gauze treatment did not reduce the abundance or diversity of yeasts in the pitchers, suggesting that insects introduce, attract, or promote the growth of yeasts. Additionally, we found that one commonly associated yeast, Candida globosa, is present internally in surface-sterilized adult W. smithii, indicating that the pitcher plant mosquito acts as a vector and may have a more complex association with this yeast. Convergently evolved pitcher plants in the genus Nepenthes exist in Southeast Asia, and our preliminary research show that there is also convergent community assembly of insect and arachnid associates between Nepenthes and Sarracenia. These results may extend to the communities of yeasts found within similar pitcher habitats on opposite sides of the planet.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P112: A modern, high-throughput workflow for biodiversity research integrating floristics, taxonomy, phylogeny, ecology and conservation

Brendan P Hodkinson1* and James C Lendemer2. 1International Plant Science Center, The New York Botanical Garden, 2900 Southern Blvd., Bronx, NY 10458-5126, 2Institute of Systematic Botany, The New York Botanical Garden, 2900 Southern Blvd., Bronx, NY 10458-5126.

The field of Systematic Biology has increasingly become fragmented in recent years, with many scientists focusing in on a small number of methods or approaches for studying biodiversity and its origins. Here we present an integrated workflow for studying the many facets of biodiversity and systematics. This workflow is currently being used to conduct a large-scale inventory of lichens in the Mid-Atlantic Coastal Plain of the United States that includes the collection and analysis of ~35,000 new specimens. Using floristic habitat sampling (FHS), specimens representing the organismal group of interest are collected from individual sites throughout the study region, and are subsequently analyzed anatomically and chemically. Specimens that cannot be identified taxonomically with anatomical/chemical analyses alone are sequenced using a multi-locus 454-based approach. Sequences from the genes of interest (in our case, mtSSU, nucLSU and nucITS) are compared with publicly-available reference sequences to determine the phylogenetic affinities of species that remain either unnamed or 'incertae sedis.' All collections are databased rapidly using modular databasing software (KE EMu). Large-scale ecological analyses are performed on database-generated taxon lists by hijacking data management software originally designed for organizing and analyzing large molecular sequence data sets. Overall results are compiled and made available on the web in the form of, e.g., checklists (for regions, sub-regions and sites), detailed taxonomic treatments, molecular sequence alignments, phylogenetic trees, and specimen-based ecological data sets. Both through these online resources and workshops/forays for the public, information is disseminated for integration into conservation and management plans to preserve biodiversity for the future.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P113: Root-associated fungi of Geum rossii and Deschampsia cespitosa at the Niwot LTER site

Zachary T Gossage1*, Sagar Yeraballi1, Katherine Suding2, Robert Sinsabaugh3, and Andrea Porras-Alfaro1. 1Biological Sciences, Western Illinois University, Macomb, IL, 2Department of Environmental Science, Policy & Management, University of California Berkeley, CA, 3Department of Biology, University of New Mexico, Albuquerque, NM.

Plant-associated fungi are known to play an important role in plant productivity and ecosystem sustainability. Endophytic fungal diversity in the alpine tundra is unknown for the majority of plant species. The main goal of this study was to identify and describe fungal symbionts associated with two alpine tundra co-dominant plants: Geum rossii and Deschampsia cespitosa. Plants were collected at Niwot Ridge long-term nitrogen fertilization experiments in Colorado. Forty-two plants were collected from four different treatments: control, nitrogen fertilized, D. cespitosa removal with nitrogen fertilization, and D. cespitosa removal with no nitrogen addition in 2008 and additional plants were collected from control plots in 2010. Roots were harvested from each treatment and stained to determine fungal colonization. Roots were surface sterilized and plated on malt extract agar with antibiotics. One hundred and ten pure cultures of endophytic fungi were isolated and identified by sequencing the internal transcribed spacer rDNA. Analysis of the sequences at 97% similarity yielded 25 contigs and 24 unique sequences. The majority of the isolates, 91%, belong to the phylum Ascomycota and the remaining to Basidiomycota and other undetermined phyla. Dominant orders included Helotiales (23%), Hypocreales (23%) and Eurotiales (21%). Within Helotiales, dominant taxa were closely related to Phialocephala fortinii and Cryptosporiopsis ericae. Within Hypocreales some common taxa included Tolypocladium and Beauveria. Dominant taxa within Eurotiales included isolates related to Penicillium sp. and Aspergillus sp. Endophytic communities showed very little plant specificity suggesting that most taxa are adapted to colonize both of these dominant species.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P114: Monitoring Fusarium complex mycelia replacement by mycopathogenic Sphaerodes using ATPtest, qPCR and HPLC

Vladimir Vujanovic. University of Saskatchewan, Saskatoon, SK, S7N 5A8 Canada.

The biotrophic mycoparasite-host interface and replacement remain poorly described, yet their understanding is important for effective biocontrol of Fusarium plant pathogens. Alcohol percentage test (APT) and real-time PCR quantification of DNA were employed to evaluate the efficiency of mycopathogenic Sphaerodes mycoparasitica mycelia to replace an array of pathogenic/toxigenic Fusaria in co-cultures. Shifts in Fusarium mycelial growth and hydrophobicity measured (APT) at the interaction zone with S. mycoparasitica indicated the degree of Fusarium mycelia replacement by the mycoparasite (P<0.05); while qPCR accurately detected DNA quantity changes in mycotoxigenic F. graminearum 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON) chemotypes challenged with S. mycoparasitica. Results from the HPLC experiments also demonstrated a considerable decrease in mycotoxin (DON, 3-ADON, 15-ADON, and ZEA) concentrations at the contact zone. Estimated shifts in the DNA quantity and mycotoxin production, assessed using qPCR and HPLC respectively, correlated with an alteration in APT values, demonstrating the effectiveness of the mycoparasite in vivo.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P115: Effect of Pleurotus tuber-regium Singer and microorganisms on degradation of unsterilized soil contaminated with cutting fluids

Clementina O Adenipekun1*, Omasan E Ejoh2, and Adeniyi A Ogunjobi2. 1Department of Botany, University of Ibadan,Ibadan,Nigeria, 2Department of Microbiology,University of Ibadan,Ibadan,Nigeria.

The study of Pleurotus tuber-regium Singer and indigenous microorganisms isolated from unsterilized soil polluted with cutting fluids were investigated over an incubation period of 0 and 2 months. The ability of these organisms to degrade Total Petroleum hydrocarbon (TPH) present in the cutting fluids, lignin content of rice straw, their enzyme activity as well as their ability to accumulate heavy metals present in the polluted soils were monitored. In all the soil samples (unsterilized soil polluted with cutting fluids (US) and unsterilized soil polluted with cutting fluids incubated with P. tuber-regium (USP), a significant increase in the nutrient content was observed with USP recording the highest. The heavy metal content of the three soil samples decreased with increase in incubation period showing that bioaccumulation of the heavy metals had occurred. The indigenous microbes alone in US accumulated heavy metals better than the indigenous microbes in USP. TPH loss (36.23%) was recorded at 10% cutting fluids concentration in USP compared to US with 19.6%. The unsterilized contaminated soil inoculated with P. tuber-regium recorded the highest lignin degradation as the polyphenol oxidase and peroxidase activity of the organisms in all the samples showed a gradual increase. The microorganisms isolated from unsterilized soil samples included Bacillus licheniformis, B. cereus, Bacillus sp., Pseudomonas aeruginosa, Pseudomonas sp., Corynebacterium sp., Neisseria sp., Trichoderma harzanium, Aspergillus niger, A. flavus, A. glaucus, Mucor sp. and Rhizopus sp.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P116: Manure gas eating microbes: Effect of fungal and bacterial biomass on biofiltration of livestock production emissions

Jason P Oliver* and Jonathan S Schilling. 320 Kaufert Laboratory, 2004 Folwell Avenue, St. Paul, MN 55108-6130.

Though specific data is only just being analyzed, scientists and regulators are beginning to recognize the significant emission of particulate matter, odor, hazardous and greenhouse gases from livestock production and manure management. Biofiltration - use of a biologically active porous media to capture and degrade gaseous pollutants - is a low-cost and adaptable mitigation technology suitable for manure storage emissions. Improving the design and management of biofilters (and ultimately adoption by producers) rests on our ability to understand the microbial ecology of the filter media; the underpinnings of biofilter effectiveness and efficiency. Studying biofilters at multiple scales, we are beginning to resolve the effect of fungal and bacterial biomass on reductions of gases generated by long-term storage of livestock manure.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P117: Morphological and molecular characterization of some Allomyces E. J. Butler strains (Blastocladiomycota) from Argentina and Brazil: preliminary results

Agostina V Marano1, Carmen LA Pires-Zottarelli2, Mónica M Steciow1, José I de Souza2, Frank H Gleason3*, Timothy Y James4, and Felipe S Chambergo5. 1Instituto de Botánica Spegazzini, calle 53 N 477, La Plata, 1900, Buenos Aires, Argentina, 2Instituto de Botânica, CP 3005, 01061-970 São Paulo, SP, Brazil, 3School of Biological Sciences A12, University of Sydney, Sydney, NSW, 2006, Australia, 4Department of Ecology and Evolutionary Biology, University of Michigan, Ann Arbor, Michigan 48109, US, 5Edificio I-1 Sala 204G, Av. Arlindo Bettio, 1000-Ermelino Matarazzo, Escola de Artes, Ciências e Humanidades, Universidade de São Paulo (USP), CEP 03828-000, São Paulo, SP, Brazil.

Allomyces is a genus of zoosporic true fungi that contains saprobic species found in freshwater and soil ecosystems worldwide. This genus possesses an isomorphic alternation of diploid sporophyte and haploid gametophyte, producing sporangia of two types (mito and meiosporangia) and male and female gametangia, respectively. There are three subgenera according to the type of life cycle exhibited: Brachiallomyces (with only sporophytic phase), Cystogenes (with sporophytic phase dominant, the gametophyte represented by a cyst) and Euallomyces (with alternation of isomorphic sporophytic and gametophytic phases). The taxonomy of this genus has been traditionally based on the presence or absence of a conspicuous gametophytic phase, the morphometric characterization of mitosporangia, meiosporangia and gametangia and the relative disposition of male and female gametangia, which are usually arranged in pairs. These characteristics are, however, extremely variable within different strains of the same species. This fact, together with the formation of natural hybrids (e.g. Allomyces x javanicus), makes the delimitation and putative identification of species challenging. In addition, recent studies have shown two clades within Allomyces with strains identified as Allomyces arbusculus in both clades, suggesting that species concepts in this genus are in need of revision. The present study aims at re-evaluating some isolates in the genus Allomyces by using morphological and molecular characters (rDNA sequences from the ITS and 28S regions). Thus far, we have characterized five isolates of A. arbusculus, A. macrogynous, A. moniliformis and A. neomoniliformis, from terrestrial environments of Argentina and Brazil. We analyzed a limited number of strains in order to provide a framework that can be use to advancing in the systematic characterization of species within this genus. This study represents the first step of a major project that attempts at involving a widespread sampling from several strains of the currently valid species.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P118: Characterizing the paracrystalline inclusion in zoospores of members of the Chytridiales (Chytridiomycota)

Peter M Letcher, Satoshi Sekimoto*, and Martha J Powell. Department of Biological Sciences, The University of Alabama, Tuscaloosa, AL 35487.

Orders within Chytridiomycota (=chytrids) have now been circumscribed as monophyletic lineages, each with zoospores exhibiting unique suites of character states. Molecular phylogenetic analyses allow us to hypothesize zoospore character evolution through tracking modifications, losses, or appearances of characters along these lineages. The presence of a paracrystalline inclusion is one zoosporic character within Chytridiomycota that has been reported only among members of the Chytridiales. We are investigating the structure and function of the paracrystalline inclusion in zoospores of the Chytridiales. Although the Chytridiales includes a variety of zoospore sub-types defining the families Chytridiaceae and Chytriomycetaceae, zoospores of all members studied with electron microscopy contain a paracrystalline inclusion. Consequently, the presence of a paracrystalline inclusion unites the Chytridiales and serves as a diagnostic character for this order within Chytridiomycota. Because paracrystalline inclusions found in organisms outside the Chytridiomycota are typically associated with the accumulation of proteins, our hypothesis is that the paracrystalline inclusion in zoospores of Chytridiales is composed of protein. We have found that number, size, and complexity of paracrystalline inclusions vary with species. The long-term aim of this study is to isolate the paracrystalline inclusion and characterize it biochemically.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P119: Exploring sporangial surface features to characterize members of the Chytriomycetaceae (Chytridiales)

William J Davis*, Martha J Powell, Peter M Letcher, and Jonathan Antonetti. Department of Biological Sciences, The University of Alabama, Tuscaloosa, AL, 35487.

Understanding species boundaries is critical to understanding biodiversity. In the past plasticity of the chytrid thallus led to the consolidation of morphologically variable isolates into a single species. Recent molecular phylogenetic analyses have often revealed great genetic diversity among isolates and validated the separation of these isolates into distinct species. It has also been revealed that morphologically similar isolates can show great genetic diversity. This is true of Chytriomyces hyalinus, one of the most commonly collected chytrids. Isolates identified as C. hyalinus based on thallus morphology and habitat form four well-supported clades in molecular analyses. These results suggest the presence of cryptic species. Characters that distinguish these potential cryptic species need to be identified in order to delineate and describe them as new species. Examination of sporangial cell surface features with scanning electron microscopy (SEM) has revealed that many of these isolates of C. hyalinus have ridge-like surfaces, which are not readily detected with light microcopy and were not observed in the original description of C. hyalinus. The purpose of this study is to explore whether sporangial surface features can be used to distinguish members in different sub-clades of the C. hyalinus complex. Thus, representatives of each of the sub-clades of the C. hyalinus complex, as well as sister clades, were examined with SEM.

Poster Session 2
Day: Monday, 16 July
Location: Omni Hotel Ballroom C
Time: 2100-2200

P120: A timescale for Oomycete evolution estimated from conserved regulators of gene expression

Jaime E Blair* and Nahill H Matari. Department of Biology, Franklin & Marshall College, Lancaster, PA 17603.

The fungal-like oomycetes are ubiquitous in nature, occupying niches in marine, freshwater, and terrestrial ecosystems. While the diversity of saprophytic oomycetes is most certainly underestimated, this group is primarily known for the important plant and animal pathogens it contains. The oldest accepted fossil evidence of biotrophic oomycetes associated with vascular plants comes from the Early Devonian Rhynie chert (approx 400 Ma); however, the affinity of fossil oomycetes to modern lineages remains unclear. Currently, genomic resources exist for a number of oomycete species, as well as close relatives within the Stramenopiles (diatoms, brown algae). The goal of this project was to use a comparative genomics approach to identify conserved regulators of gene expression within each of these genomes, and use Bayesian techniques to co-estimate phylogeny and divergence times. We focused primarily on genes involved in key processes such as RNA silencing and chromatin modification, as well as core eukaryotic transcription factors. Oomycete genomes do contain the necessary complement of genes for RNA silencing, including orthologs of Dicer, Argonaut, dsRNA-binding proteins, and an RNA-dependent RNA polymerase; other researchers have shown that these genes are expressed during various life stages in Phytophthora infestans. Oomycetes also possess a full array of genes for histone modification, including deacetylases, acetyltransferases, methyltransferases, and chromodomain-containing proteins. Surprisingly, no homologs to canonical eukaryotic DNA methyltransferases could be identified in the genomes of oomycetes or in the outgroups. Bayesian divergence times were estimated from a dataset of 53 proteins using conservative calibrations from the diatoms and the Rhynie chert oomycetes. The sensitivity of estimated divergence times to model specification on the prior parameters will also be discussed.