Inoculum 56(4) - Mycological Society of America

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MSA ABSTRACTS trol and experimentally N-fertilized treatments at three forest sites found along a N deposition gradient in the Chicago region. Field surveys of sporocarps in treatment plots have identified over 90 ECM fungal species across the three sites since 2003. Over 200 of these collections were used to develop a reference database of terminal restriction fragment length polymorphisms (T-RFLP) for the identification of ECM collected belowground. In 2004, we conducted morphological and T-RFLP analyses of ECM collected from soil cores from each treatment plot. Early results indicate that the ECM communities in these sites are rich and potentially vulnerable to N increase. Over 130 likely species of ECM fungi have been identified from over 5000 oak root tips examined. Species richness estimates indicate that significantly fewer numbers of ECM species are found on oak roots in N fertilization treatments at two of three sites. We will continue to monitor ECM community responses to N fertilization over the next two years and also examine how the composition of these communities relates to their function within the context of N deposition. contributed presentation Badalyan, Suzanna M.*, Garibyan, Narine G. and Sakeyan, Carmen Z. Dept. of Botany, Yerevan State University, Aleg Manoogian St., 375025, Yerevan, Armenia. badalyan_s@yahoo.com. Culture collection of Basidiomycetes fungi at the Yerevan State University (Armenia). Establishment and maintenance of Culture Collection of macroscopic fungi (Basidiomycetes) at the Yerevan State University are the way of preserving biodiversity of mushrooms and extending fungal genetic and biotechnological research in Armenia. Study of mushroom cultures can also be valuable in obtaining novel bio-pharmaceuticals and functional food additives with health-enhancing effect. Presently, the Collection comprises around 210 living strains of 60 mushroom species. They were mostly isolated in Armenia and obtained from other institutions. Among them, 35 species and 197 strains possess known medicinal properties. The Flammulina velutipes, Pleurotus ostreatus and Coprinus spp. collections are represented by a wide eco-geographical diversity of strains. ITSrDNA nucleotide sequence analyses of collected 25 species and 105 strains were carried out together with international collaborators. The project of genetic identification of Armenian medicinal mushrooms is currently realizing. Further extension of taxonomic, ecological and geographical diversity of species/strains and their genetic identification, as well as digitalization and creation of information DataBase are in progress. The catalogue of Culture Collection will be available soon. This research is supported by NATO (#FEL.RIG.980764) and DAAD (#548.104401.174) grants. poster Badalyan, Suzanna M. 1 * and Kües, Ursula 2 . 1 Dept. of Botany, Yerevan State University, Aleg Manoogian St., 375025, Yerevan, Armenia, 2 Section Molecular Wood Biotechnology, Institute of Forest Botany, Georg-August University, Büsgenweg 2, D-37077 Göttingen, Germany. badalyan_s@yahoo.com. Mycelial morphology and growth characteristics of wood-related coprinoid mushrooms. Around 80 species of the traditional genus Coprinus (Coprinoid mushrooms) have been observed on wooden material. Mycelial micro-, macromorphology and growth characteristics of xylotrophic species Coprinus comatus, Coprinellus angulatus, C. bisporus, C. curtus, C. disseminatus, C. domesticus, C. ellisii, C. micaceus, C. xanthothrix, Coprinopsis atramentaria, C. cinerea, C. cothurnata, C. gonophylla, C. radians, C. romagnesiana, C. scobicola, C. strossmayeri and Parasola plicatilis have been studied. Cultures were grown on Malt- Extract Agar (MEA), Potato-Dextrose Agar (PDA) and Glucose-Peptone Agar (GPA) at 25 ∞C and pH 6. Growth rates and growth coefficients were highest on MEA (up to 85 mm and above 20, respectively), then PDA and GPA. Macromorphological characteristics were described after 10 days of growth. Oval and round shape clamps occur in most of the Coprini. Clamps were not found in some Coprinellus species. Material for micromorphological investigations was obtained by slide cultures. Hyphal loops were particularly formed in Coprinellus species. Arthroconidia were often observed, whereas blastic sporulation was rare. Chlamydospore formation is also typical for Coprini. Mycelial cysts, microfilaments and crystals were detected in some species.Thanks DAAD, NATO and Deutsche Bundesstiftung Umwelt for financial support. poster Bahl, Justin, Jeewon, Rajesh* and Hyde, Kevin. Dept. Ecology & Biodiversity, The University of Hong Kong, Hong Kong, SAR, China. jbahl@hkusua.hku.hk. Intergeneric relationships of Linocarpon and Neolinocarpon: does phylogenetic analysis support the generic delineation? Species from the genera Linocarpon and Neolinocarpon are common saprobic fungi found in subtropical to tropical regions and mainly occurs on monocotyledonous hosts. Both genera are often found on the same host. Based on morphology these genera share many similarities and have resulted in difficulties in assigning taxa. The most significant delineating characters are ascomata position and morphology. Based on parsimony and likelihood analyses of multi-locus partial sequences derived from nuclear encoded ribosomal DNA, beta-tubulin and RNA polymerase regions from fresh and dried herbarium material, an attempt has been made to assess which morphological characters are phylogenetically significant for generic delineation or whether the genera should be circumscribed under the priority name, Linocarpon. Analysis confirmed that the two genera are not monophyletic and indicates parallel evolution of morphological and ecological 8 Inoculum 56(4), August 2005 characters. Results are discussed in relation to the significance of morphological characters currently used in the taxonomy of Linocarpon and Neolinocarpon. contributed presentation Barnes, Irene*, Wingfield, Michael J. and Wingfield, Brenda D. Department of Genetics, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria 0002, South Africa. irene.barnes@fabi.up.ac.za. Development of microsatellite markers for the red band needle blight pathogen Dothistroma septosporum using two different isolation methods. Very little is known regarding the population biology of Dothistroma septosporum, one of the most important pathogens of plantation grown pines in the southern hemisphere. Thus, twelve sets of microsatellite markers have been developed to study the population dynamics of this pathogen. Two techniques, ISSR-PCR and FIASCO enrichment were used to screen for microsatellite rich regions. ISSR-PCR was effective in locating many microsatellite sites. However, after the necessary genome walking, many of the microsatellites were found to be redundant artifacts of the initial primers used. With FIASCO, variable success was observed depending primarily on the primer combination used in the enrichment. In one screen, 57 % of the clones contained microsatellites, in others, none were found. From a total of 22 primer pairs, 11 were found to be polymorphic amongst isolates of D. septosporum. An additional primer was polymorphic between D. pini and D. septosporum and can be used for further diagnostic purposes within populations. Cross-species amplification was successful in D. pini, D. rhabdoclinis and Mycosphaerella dearnessi. Future studies using these primers will focus on gaining an improved understanding of the population structure, genetic diversity, gene flow and the genetic relatedness between different populations of these important tree pathogens. poster Barnes, Irene 1 *, Crous, Pedro W. 2 , Wingfield, Michael J. 1 and Wingfield, Brenda D. 11 Department of Genetics, Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria, South Africa, 0002, 2 Centraalbureau voor Schimmelcultures (CBS), Fungal Biodiversity Centre, P.O. Box 85167, 3508 AD Utrecht, The Netherlands. irene.barnes@fabi.up.ac.za. Multigene phylogenetic analyses reveal that Dothistroma septosporum and D. pini represent two distinct taxa and a serious threat to pine forestry. The sudden increase in severity of the red band needle blight disease in the U.K., Canada and parts of Europe where Dothistroma has been present for decades is a matter of great concern. Although the etiology of the disease is well known, phylogenetic and population level relationships amongst isolates of the fungus are poorly understood. We have thus constructed multigene phylogenies for isolates of Dothistroma from 13 different countries. These have illustrated that the isolates are separated into two distinct lineages representing two discrete species and supported by morphological differences. The one species, referred to as D. septosporum occurs worldwide and infects over 30 species of pines. It is the major cause of the serious blight disease plaguing Pinus radiata plantations in New Zealand, Chile and other Southern Hemisphere countries The second species, D. pini, is restricted in its distribution to the North Central United States where it causes a serious disease on exotic P. nigra. A simple ITS-PCR-RFLP is also presented that allows accurate and rapid distinction between the two species. contributed presentation Baroni, Timothy J. 1 *, Lindner Czederpiltz, Daniel L. 2 , Lodge, D. Jean 3 , Hofstetter, Valérie 4 and Franco-Molano, Ana Esperanza 5 . 1 Department of Biological Sciences, State University of New York, College at Cortland, Cortland, NY 13045, USA, Center for Forest Mycology Research, USDA Forest Service, Forest Products Laboratory, One Gifford Pinchot Dr., Madison, WI 53726-2398, USA, Center for Forest Mycology Research, USDA Forest Service, P.O. Box 1377, Luquillo, PR 00773-1377, USA, 4 Botany Department, Duke University, Durham, NC 27708-0338, USA, 5 Laboratorio de Taxonomía de Hongos, Instituto de Biología, Universidad de Antioquia, A.A.1226, Medellín, Colombia. baronitj@cortland.edu. Arthrosporella, a recently rediscovered neotropical genus, is phylogenetically related to Termitomyces in the Lyophylleae. In 1996, Sharon Cantrell and TJB collected an odd nail-shaped agaric covered with dark brown conidia in Puerto Rico; that collection could not be named at that time. Over the next 9 years of intense collecting, this odd arthrospore-producing species was collected only three more times in Puerto Rico, but also twice in the Dominican Republic. TJB realized this was a new species of Arthrosporella, originally described by Rolf Singer as a monotypic genus from Argentina. A second, distinctly different and new conidia producing agaricoid species was found just recently in Colombia by AEFM, and we now know of two other collections of this taxon from the Dominican Republic by Egon Horak and TJB. Very recently (August 2004) a third new, and completely different arthrosporeproducing agaricoid species was found by TJB, DJL and CDL, fruiting abundantly in the cloud forest on the highest peak in Belize (Doyle’s Delight). All of these new taxa possess siderophilous granules in the basidia. Phylogenetic analysis of nLSU indicates, with significant support, the monophyly of Arthrosporella with Termitomyces (Lyophylleae) and suggests a sister relationship between these two genera. All collections of Arthrosporella appear to be saprotrophic and not termitophilous, thus perhaps indicating a closer relationship with Podabrella, which is also in this branch of the Lyphylleae clade. poster Continued on following page
Barrett, Luke G., Thrall, Peter H., Burdon, Jeremy J. and van der Merwe, Marlien M.* Centre for Plant Biodiversity Research, CSIRO – Plant Industry, GPO Box 1600, Canberra ACT 2601, Australia. marlien.vandermerwe@csiro.au Pancontinental patterns of genetic variation in the rust fungus Melampsora lini. The Linum marginale-Melampsora lini plant-pathogen interaction is endemic to Australia and has been a focus of epidemiological and coevolutionary studies for more than a decade. Considerable variation for both host resistance and pathogen virulence has been shown at a range of spatial scales from the local to the continental. Here we report on a study using AFLP and SSR markers to examine pancontinental patterns of genetic variation in 102 clonal lines of M. lini representing 35 populations. Molecular marker genotypes partition all of the isolates into two major lineages, and in combination with subsequent sequencing of beta-tubulin and elongation factor genes suggest a possible hybrid origin for one of the lineages. Subsequent comparison with data on phenotypic variation for virulence in a subset of these isolates also demonstrates striking differences between the two lineages in terms of pathogenicity on the host L. marginale. Molecular genetic variation within the lineages was very limited, and within populations both AFLP and SSR markers regularly failed to distinguish among several lines with different pathotypes. These results are important for developing an understanding of how pathogen virulence might evolve within natural populations. poster Barrow, Jerry R.*, Lucero, Mary L., Osuna-Avila, Pedro, Reyes-Vera, Isaac and Aaltonen, Ronald E. USDA-ARS Jornada Experimental Range, Las Cruces, NM 88003, USA. jbarrow@nmsu.edu. Hybrid vigor, structural modification and enhanced plant performance induced by symbiotic fungi-plant interactions. Hybrid vigor, structural modification and enhanced plant performance induced by symbiotic fungi-plant interactions. Communities of symbiotic fungi are intrinsically integrated with single cells, tissues and organs of desert plants. They are obligately associated with living tissues and generally cannot be isolated and cultured separately. Symbiotic fungi indigenous to native desert plants were transferred, using cell culture methods, to non-host plants and reciprocally transferred between native grasses. Crop and native plants, with recipient fungi have astounding and increased levels of vigor. Up to five-fold increases in root and shoot biomass and substantial morphological changes and reproductive potential were obtained. Enhanced chlorophyll and phosphorous content were common responses. Once integrated, fungi were intimately interfaced with the plasmalema of the new host plant and were ecto-cytoplasmically inherited. This technology accesses a previously unexplored source of genetic variability within native ecosystems with the potential of immediate transfer to improve tolerance of native and crop plants to stress, disease and pests. Symbiotic fungal transfers provides a powerful alternative for genetic improvement of plants. contributed presentation Bartnicki-García, Salomón. Microbiology Dept., CICESE, Ensenada, Baja California, 22860, México. bartnick@cicese.mx. Introductory remarks: Current perspectives in hyphal morphogenesis. Hyphal morphogenesis is clearly the most basic developmental process in mycology. The origin of the fungal kingdom could be traced to the “invention” of hyphal morphogenesis, i.e. the ability of a cell to form long tubular walls by tip growth. Not surprisingly, over the years, fungal biologists have become increasingly interested in elucidating the cellular and molecular basis of hyphal morphogenesis. The problem revolves basically about understanding the mechanism(s) responsible for polarized growth of the cell wall. A number of structural and molecular players have been identified but the mechanism cannot be attributed to a single gene or protein. Polarized secretion probably requires a specific concerted action between cytoskeleton and secretory vesicles to produce a pattern of vesicle discharge that would generate cells with a profile described by the hyphoid equation: y = x cot (xV/N). This symposium on Advances in Fungal Morphogenesis will cover developments in different fronts: a new way of probing the surface properties of the hyphal wall (S. Kaminskyj), assessing the importance of protein glycosylation in the maintenance of hyphal growth (B. Shaw), and analyzing the role of the Spitzenkörper in apical growth (M. Riquelme). symposium presentation Baucom, Deana L.*, Bruhn, Johann N. and Mihail, Jeanne D. Division of Plant Sciences, University of Missouri, Columbia MO 65211, USA. dlba3c@mizzou.edu. Armillaria species involved in Missouri Ozark forest decline. In the Missouri Ozark Mountains, Armillaria spp. contribute to oak decline. To investigate the distributions and roles of Armillaria spp., 142 isolates collected in 2002 from 40 plots were identified. Amplification of IGS1 followed by restriction with AluI identified 121 isolates as A. mellea (52 %), A. gallica (38 %), and A. tabescens (10 %). Two new RFLP patterns were found to represent 41 % of A. mellea and 17 % of A. gallica isolates. Isolates yielding these patterns are currently being characterized by sequence analysis of IGS1. Major hosts were dogwood (44 %), red oaks (18 %), and white oaks (7 %). Armillaria mellea predominated on both white and red oaks; A. mellea and A. gallica occurred equally on dogwood. Recently killed trees provided 28 % of study isolates. Armillaria was almost universally present on recently killed trees. Red oaks, dogwood, and white oaks provided 38 %, 32 %, and 10 % of recent mortality isolates, respectively. Nearly all recent oak mortality yielded A. mellea. Recent dogwood mor- MSA ABSTRACTS tality yielded A. gallica (54 %) more frequently than A. mellea (38 %). Ozark oak decline affects red oaks and dogwoods most heavily. The Armillaria sp. which most commonly contributed to oak mortality on our plots in 2002 was A. mellea, though A. gallica was more commonly recovered from recent dogwood mortality. We found little evidence of mortality caused by A. tabescens. poster Beeson, Esther 1 , Beltz, Shannon 2 , Klich, Maren 2 and Bennett, Joan W. 1 * 1 Tulane University, New Orleans, LA, USA, 2 Southern Regional Resource Center, New Orleans, LA, USA. ebeeson1@cox.net. Sclerotial production in Aspergillus flavus varies with temperature and nitrogen source. Twenty strains of Aspergillus flavus from the culture collection of the Southern Regional Research Laboratory, New Orleans, LA, were grown on defined and complex media at 25 o C and 37 o C for one week. Colonies were screened for sclerotial production as well as other colony characters including production of conidia, floccose hyphae, and mycelial pigment. When grown on complex media with yeast extract as the nitrogen source, 9 of the strains produced sclerotia at 25 o C and 11 produced sclerotia at 37 o C. When grown on a defined medium with nitrate as nitrogen source, 8 of the strains produced sclerotia at 37 o C and 5 produced sclerotia at 25 o C. When ammonium was used as the nitrogen source, only one strain produced sclerotia at 37 o C and there was no sclerotial production at 25 o C. Sporulation was sparse or absent for all strains grown on ammonium at 37 o C. When microarrays of the A. flavus genome become available, these data will be useful in designing conditions for RNA isolation in order to probe the genes involved in sclerotial formation. poster Bennett, Chandalin 1 *, Newcombe, George 1 and Aime, M. Catherine 2 . 1 Department of Forest Resources, University of Idaho, Moscow, Idaho 83844, USA, 2 USDA ARS Systematic Botany and Mycology Lab, Beltsville, Maryland 20705, USA. benn4449@uidaho.edu. Regional studies of Melampsora on Salix in the Pacific Northwest. Melampsora epitea Thuem. is a species complex that represents all willow rusts in N. America. To better understand how much host-specialization and genetic diversity exists in the Pacific Northwest (PNW), a large-scale host-range inoculation study was performed along with ITS sequencing, morphological analysis, and a two-year field survey. Distinct host-specificity was shown for three different Melampsora isolates inoculated on nearly equal sets of 440 willows. Less than 20 percent of the plants in each experiment were susceptible to the inoculum and greater than 15 percent of those showed some signs of resistance. The genetic sequencing resulted in four distinct clades, the largest of which likely represents a complex in this region. The other three clades were strongly divergent from the complex. There was up to three genetically distinct rusts present in a given geographic location and four or more genetically distinct rusts on a given Salix sp. There was found at most two distinct rusts on the same species from the same population. It’s evident that willow rusts are incredibly diverse in the PNW. Every experimental inoculum was specialized in its host range and the genetic diversity was spread across four unique and highly divergent clades. symposium presentation Bennett, Joan W. Tulane University, New Orleans, LA 70118, USA. jbennett@tulane.edu. Industrial mycology: from Takamine’s diastase to TIGR’s database. Industrial mycology has its roots in numerous food fermentations that were developed early in human history. Of these, the Japanese koji process, utilizing Aspergillus oryzae , is of particular interest because the first microbial enzyme ever patented was a secreted diastase (amylase) isolated from A. oryzae by Dr. Jokichi Takamine, a Japanese chemist working in the U.S.A during the late nineteenth century. In the first half of the twentieth century, several filamentous fungi were harnessed for the production of additional commercial enzymes and for organic acids. Although the discovery of penicillin and the subsequent golden age of antibiotics transformed fermentation technology, the molecular biology of industrial molds lagged behind that of model species. Now, in the twenty-first century, genomic research has ushered in a new era of understanding. Genome projects for several Aspergillus species are completed or underway, revealing new insights into secondary metabolism, carbon and nitrogen regulation, and other genetic control mechanisms. aymposium presentation Berbee, Mary L. 1 *, James, Tim Y. 2 , Longcore, Joyce E. 3 , Stajich, Jason 4 and Vilgalys, Rytas J. 2 . 1 Dept. of Botany, University of British Columbia, V6T 1Z4 Canada, 2 Dept. of Biology, Duke University, Durham, NC 27708 USA, 3 Biological Sciences, University of Maine, Orono, ME 04469-5722 USA, 4 Dept. of Molecular Genetics and Microbiology, Duke University, Durham, NC 27708, USA. berbee@interchange.ubc.ca. What makes a fungus? Fungal-specific genes from EST libraries of the basal fungi Batrachochytrium dendrobatidis and Mortierella verticillata. The Chytridiomycota and Zygomycota include ancient fungal lineages that may have originated hundreds of millions of years before plants invaded land. We looked for shared genes that distinguish the fungi from other kingdoms through comparison of expressed sequence tag (EST) libraries, from a chytrid, Batrachochytrium dendrobatidis (1588 ESTs, average length, 572 nucleotides) and Continued on following page Inoculum 56(4), August 2005 9
Page 1 and 2: Newsletter of the Mycological Socie
Page 3 and 4: Cordyceps Diversity in Korea Cordyc
Page 5 and 6: From the President’s Corner … D
Page 7: of fungal glucan synthesis. This in
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Upadhyay, Srijana, Long, Melissa an
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MYCOLOGIST’S BOOKSHELF This issue
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MYCOLOGIST’S BOOKSHELF Previously
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Ascomycota of Sweden www.umu.se/myc
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inoculum The Newsletter of the Myco
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An Invitation to Join MSA THE MYCOL
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