Inoculum 56(4) - Mycological Society of America

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MSA ABSTRACTS of AM colonization in C. barbinervis decreased when C. barbinervis grown where cucumber was dominant. These suggest that plant species affect selection of AM fungi in neighbor plants. Influence of plant root exudates on AM colonization was discussed here. symposium presentation Kuga-Uetake, Yukari. Faculty of Agriculture, Shinshu University, Minami-Minowa, Nagano 399-4598, Japan. ykuga@shinshu-u.ac.jp. Diversity and similarity in the structure of mycorrhizas. Most plant species establish symbiotic organs called mycorrhizas, structures formed between plant roots and fungal hyphae. The organs are sites of exchange of nutrients between two organisms, and to date, seven types are recognized based on their structure. Except for ectomycorrhiza, arbuscular, ectendo-, monotropoid, arbutoid, ericoid, and orchid mycorrhizas establish intracellular hyphae. The intracellular hyphae are, however, always separated from host cells by host membrane continuous with the plasma membrane and cell wall materials derived from the host. Ecto, ectendo, arbutoid, and monotropoid mycorrhizas develop hyphal layers intercellularly, which is called a Hartig net, and around the root tip called a mantle. Host microtubule (MT) arrays change accompanying intracellular fungal colonization in arbuscular, orchid, ectendo- and monotropoid mycorrhizas. MTs were always closely associated with the membrane surrounding the fungal hyphae. Recently in ecto-, ectendo- and monotropoid mycorrhizas, modified cortical MTs were also observed for the first time in host cells adjacent to Hartig net hyphae. The seven mycorrhizal types are distinctive in the structures but share a common feature of the presence of host MTs on membranes closely associated with fungal structures. symposium presentation Kunishi, Ayako* and Hashimoto, Yasushi. Agro-environmental Science, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan. yhashi@obihiro.ac.jp. Mycorrhizal colonization and structure of Pyrola incarnata Fischer growing in a Japanese larch forest. Pyrola L. is known to form arbutoid mycorrhiza, but details of morphology and ecology of Pyrola mycorrhizas are not clear. To investigate the seasonal change of quality and quantity of Pyrola mycorrhizas, the roots of Pyrola incarnata Fischer were collected from a Japanese larch (Larix kaempferi Carr.) forest. These mycorrhizas were divided into morphological types by details of mycorrhizal surface. Each type was sectioned for observation of mycorrhizal fungal structures. Some dominant types of Pyrola mycorrhizas and ectomycorrhizas of L. kaempferi collected from a forest site were analyzed for restriction fragment length polymorphisms (RFLP) of the amplified internal transcribed spacer (ITS) region in nuclear rDNA. Pyrola mycorrhizal colonization rate and number of types were increased from spring to summer. A total of 11 types were present in Pyrola mycorrhizas. Observations of the sections of mycorrhizas showed that two types have only intracellular hyphe with no organized fungal sheath and one type have only the sheath. The other types were typical arbutoid mycorrhiza that have organized sheath, Hartig net and intracellular hyphe. ITS-RFLP pattern of the most dominant type of Pyrola mycorrhizas had the same pattern of ectomycorrhizas of L. kaempferi. Thus the fungi that colonized on these dominant types were identified as Thelephoraceae by ITS sequences. poster Kurihara, Yuko 1 *, Ogawa, Yoshio 2 , Degawa, Yousuke 3 and Tokumasu, Seiji 1 . 1 Sugadaira Montane Res. Cent., Univ. of Tsukuba, 1278-294, Osa, Sanada, Nagano 386-2201, Japan, 2 Coll. of Pharmacy, Nihon Univ., 7-7-1, Narashinodai, Funabashi, Chiba 274-8555, Japan, 3 Kanagawa Pref. Museum of Natural History, 499, Iryuda, Odawara, Kanagawa 250-0031, Japan. kurihara-yuko@nite.go.jp. A proposal for the division of the order Kickxellales based on the comparison with its related orders. The order Kickxellales Benjamin 1979 has been regarded as a close relative of Harpellales, and most of the species are saprobes and inhabit in soil or on dung of omnivorous or herbivorous mammals. Though the current classification system of the order lacks consistency due to the inclusion of newly added taxa after Benjamin (1959), no comprehensive taxonomic studies have been made after his work. To construct a more consistent taxonomic system of Kickxellales, the following strategy was used. First, all kickxellalean genera were classified into groups based on the optical microscopic morphology. Second, these groups were evaluated based on the septal ultrastructure and 18S and 28S rDNA sequence analysis, respectively. From these results, a taxonomical conclusion was drawn. Three groups; Coemansia group, Spiromyces group, and Ramicandelaber group were recognized in Kickxellales, and they were morphologically and phylogenetically distinct enough from each other and from any other fungal groups including Dimargaritales and Harpellales. Thus, each of the three groups would be regarded as an independent order, that is, Kickxellales sensu Benjamin 1979 would be divided into three orders. This treatment makes the range of Kickxellales recur to the original one that has been stretched out gradually. symposium presentation Kurihara, Yuko 12 *, Machida, Ryuichiro 3 and Fukui, Makiko 3 . 1 Dept. of Biotech., Nat. Inst. of Tech. and Eval., Kazusakamatari, Kisarazu, Chiba 292-0818, Japan, 2 Mycol. & Metabol. Divers. Res. Cent., Tamagawa Univ. Res. Inst., Machida, Tokyo 194-8610, Japan, 3 Sugadaira Mont. Res. Cent., Univ. of Tsukuba, Osa, Sanada Nagano 386-2201, Japan. kurihara-yuko@nite.go.jp. Fungi isolated from proturans under rearing. 34 Inoculum 56(4), August 2005 Protura is one of the most primitive hexapod orders, and all the species live in soil. Fungal parasites of soil arthropods including proturans were scarcely investigated while entomopathogenic fungi of Pterygota have been relatively fully studied and already applied for industry. To develop a new isolation source of industrially useful fungi, we studied fungi isolated from Baculentulus densus (Acerentomidae, Protura). Proturans were extracted from litter and soil collected at Shinko-ji, Sanada, Nagano, Japan with a Tullgren funnel, and kept in rearing containers with a small amount of litter in laboratory. Fungi were isolated from dead/intact bodies of proturans after rearing. As a result, Acremonium kiliense, Lecanicillium psalliotae and other Verticillium sensu lato species, and Conidiobolus coronatus dominantly appeared from them. A strain of A. kiliense showed an antifungal activity. Lecanicillium psalliotae has been shown to be phylogenetically close to Isaria spp. (Luangsa-ard et al. 2004), and C. coronatus is known as a biological-control agent. These results suggest that proturans and other soil arthropods would provide us industrially important fungi as Pterygota. poster Kwan, Hoi-Shan*, Chum, Wing Yan, Bian, Xue-Lin, Xie, Wei-Jun, Ng, Zhang, Leung, Grace, Sze Wan. Department of Biology and Food and Nutritional Sciences Programme, The Chinese University of Hong Kong, Hong Kong SAR, China. hoishankwan@cuhk.edu.hk. Gene expression profiles of Shiitake mushroom Lentinula edodes revealed by Differential Display, cDNA Microarray and Serial Analysis of Gene Expression (SAGE). Fruit body development is an important area in mushroom biology and has recently been studied at the molecular level. We aim to characterize gene expression profiles during fruit body development of Shiitake mushroom Lentinula edodes. First, we used the differentially display method RNA fingerprinting with arbitrarily primed polymerase chain reaction (RAP) to isolate genes differentially expressed during fruit body development. RAP is powerful in isolating gene fragments but requires tedious down-stream works to isolate full-length for further analysis. Over 100 genes were isolated and sequenced. Fifteen were studied further. Second, RAP products were used as probes to hybridize to cDNA macroarray membranes to identify and sequenced over 100 differentially expressed genes. Third, the cDNA clones from a cDNA library of primordium were randomly picked and sequenced to generate over 500 unique Expressed Sequence Tags (ESTs). Differential expressions of ESTs were analyzed by dot-blot hybridization and cDNA microarray analysis using total cDNA from mycelium and primordium as probes. From the above analysis, only relative levels of differentially expressed genes were obtained. To determine the proportion of each mRNA among total transcripts in mycelium and primordium, we used Serial Analysis of Gene Expression (SAGE) which reports the number of transcripts for each gene. About 20,000 transcripts were counted from five developmental stages and 110 genes could match to our ESTs. Examination of the kind of genes abundantly and differentially expressed in primordium indicated that at the initiation of fruit body, the following occurred: (1) specific sets of genes are expressed in primordium, (2) mycelium-specific genes are suppressed, (3) different sets of structural proteins appear in different stages, (4) protein turn-over increases, (5) protein synthesis increases, and (6) specific signal transductions occur. Gene expression profiles revealed by different approaches were compared and were generally consistent. contributed presentation Lam, Wing Hon 1 *, Taylor, Paul W.J. 2 , Jeewon, Rajesh 1 and Hyde, Kevin D. 1 1 Centre of Research in Fungal Diversity, Department of Ecology & Biodiversity, University of Hong Kong, Hong Kong SAR, China, 2 Bio Marka, Joint Centre for Crop Innovation, Institute of Land and Food Resources, University of Melbourne, Vic, 3010, Australia. winghon@hkusua.hku.hk. Molecular phylogeny of the pathogenic falcate-spored Colletotrichum species. Colletotrichum and its teleomorph Glomerella are important plant pathogens that cause a disease known as anthracnose. The economic impact due to the damage of crops by this disease is huge. Species concepts in Colletotrichum have been mainly based on morphological and cultural criteria, until recently molecular phylogenetics were utilized to study the relationship of some of the agricultural important species e.g. C. gloeosporioides, C. acutatum, C. musae. In our study, we focused on the pathogenic falcate-spored Colletotrichum species, including those infecting grasses and the non-graminicolous ones. The morphological characters in these species are often highly variable and have caused a lot of confusions in the differentiation of species. Using the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA and beta-tubulin (tub2) gene, we tried to elucidate the interspecific relationships between the selected isolates. We have also examined the type materials of C. capsici, C. caudatum, C. falcatum and C. trichellum. Fresh isolates were also obtained from the original collection locations and epitypes/lectotypes were designated in order to stabilize the application of species names. poster Landis, Frank C. 1 * and Gargas, Andrea 2 . 1 University of Akron, Akron OH, USA, 2 University of Wisconsin-Madison, Madison, WI, USA. flandis@uakron.edu. ITS2 secondary structure defines species-specific probe regions for fungi. We designed prototype DNA microarrays including 183 ITS2 rDNA sequences from 162 fungal species known to inhabit soil, with representative Continued on following page
species from each of the five Eumycota phyla plus Oomycota. Probes were based on 20 nt segments of each ITS2 sequence, with microarrays containing all possible 20 bp segments. Arrays were designed to detect multiple fungal species from soil samples. Initial results provided substantial insight into designing eukaryotic rDNA microarray detectors, as these microarrays were compromised by two design limitations: 1) 55% of the probes were shared among two or more species. This is understandable, as ITS2 regions contain substantial phylogenetic information, suggesting that a number of 20-nt oligonucleotide segments should be similar or identical among species. 2) Most seriously, prototype design also proved susceptible to spoofing, responding to fungi not represented on the array. For example, one test species hybridized with single-copy probes from seven different species, although supposedly it was not represented in the array. Since most soil fungal species are unidentified, probes designed to detect known taxa must also have safeguards against responding to unknown taxa. Fortunately, one significant result potentially overcomes both limitations. Analysis of probe duplications against 119 of the array sequences showed that unique sequences tended to occur most often around nt position 40 of ITS 2, where 67% of probes were unique, and duplicates were confined to near relatives. Position 40 corresponds precisely with loop 2 of the ITS2 folding structure, a folding structure repeated in all 119 sequences. Our findings suggest that probes based on loop 2 sequences would a priori be close to taxon-specific and therefore resistant to spoofing. More generally, analysis of secondary structure folding patterns in rapidly evolving sequences holds promise for the design of taxon-specific oligonucleotide probes. poster Landolt, John C. 1 *, Slay, Michael E. 2 and Stephenson, Steven L. 3 1 Dept. of Biology, Shepherd University, Shepherdstown WV 25443, USA, 2 Ozark Highlands Office, The Nature Conservancy, Fayetteville AR 72701, USA, 3 Dept. of Biological Sciences, University of Arkansas, Fayetteville AR 72701, USA. jlandolt@shepherd.edu. Dictyostelium rosarium and other cellular slime molds from Ozark caves. Samples of “soil” material were collected from 33 caves in Arkansas, Missouri and Oklahoma. These samples were processed in the laboratory using standard isolation procedures for dictyostelid cellular slime molds. These organisms were recorded from 18 of the 33 (55%) caves. In addition to the fairly cosmopolitan species Dictyostelium mucoroides, Polysphondylium pallidum and P. violaceum, five other species were recovered, including numerous isolates of D. rosarium from 12 different caves. Based upon these data and an earlier study of West Virginia caves, D. rosarium appears to have a preference, or at least a particular tolerance, for cave environments. In general, the pH values of soil samples from Ozark caves were more acidic than those from the West Virginia caves sampled previously. This project was supported in part by the National Science Foundation, the University of Arkansas, Shepherd University, and The Nature Conservancy. poster Landolt, John C. Dept. of Biology, Shepherd University, Shepherdstown WV 25443, USA. jlandolt@shepherd.edu. Studies of Alaskan cellular slime molds. In the 1990’s, the results of several studies of cellular slime molds (CSM) of high-latitude regions of Alaska were published in the journal Arctic and Alpine Research. Additionally, a number of other studies were carried out and one project is still ongoing. This presentation summarizes the results of this work on Alaskan CSM, published and unpublished. Although occurring at low levels of species richness in high latitudes regions of western and central Alaska, measured densities of CSM sometimes rival those of lower latitudes. One probable new species has been recovered, and some interesting patterns of ecological succession in CSM communities are suggested by the data obtained from the various study sites. This work has benefited from the efforts of Dr. S. L. Stephenson, logistical support and funding from Dr. G. A. Laursen (UAF/National Park Service research grants Nos. PX9830-93-062, PX9830-92-385, PX9830-0-0451, PX9830-0-0472, and PX9830-0-0512) and from contributions provided by a number of students and technicians, particularly Woody Wingate and Bess Morrison. Dr. Glen Juday was instrumental in setting up the study that allowed data to be collected from the Columbia Glacier region. Thanks also to personnel of the U.S. National Park Service, funding provided by the National Geographic Society (NGS grant #3974- 88), and logistical support from Shepherd University. symposium presentation Laursen, Gary A. 1 *, Horak, Egon 2 and Taylor, D. Lee 3 . 1 UAF, Inst. of Arctic Biology, P.O. Box 756100, 305A Bunnell Bldg., Fairbanks, AK 99775, USA, 2 ETH Zentrum, University of Zurich, Zurich, SZ, Switzerland, 3 UAF, Inst. of Arctic Biology, 311 Irving I, Fairbanks, AK, 99775, USA. ffgal@.uaf.edu. Galerina patagonica Singer from Gondwanian Mainland AU and NZ, their Subantarctic Islands, and Patagonia. Twenty-eight collections (Galerina patagonica Singer) were examined from the Subantarctic Islands (SAIs) of Macquarie (540 S., AU), Campbell (520 S, NZ) and Auckland (500 S., NZ), but not yet recorded from other SIAs, and from mainland NZ and AU. SAI substrates included peaty soil, vascular plant litter of Poa foliosa, Stilbocarpa polaris, Pleurophyllum hookeri, Dracophyllum longifolium, D. scoparium, Metrosideros umbellata and mosses. The biodiversity of island agaric floras show affinities with Patagonia (S.Am.) 2700 km NE. G. patagonica Gondwanian distribution strongly supports long-distance wind and/or bird dispersal mechanisms. To investigate the systematic and phylogeography of MSA ABSTRACTS G. patagonica, the internal transcribed spacer (ITS) was sequenced in addition to part of the RPB1 gene in a subset of 13 specimens. Data analyses revealed two clades within G. patagonica that were congruent across the two genes, robust to methods of phylogenetic inference and strongly supported. We suggest the presence of two cryptic species within the currently recognized species. Clade 1 was found in material from both mainlands as well as Auckland and Macquarie Islands. Clade 2 was found on all three Subantarctic islands, but not on the two mainlands. Identical sequences were often found in multiple localities indicating recent long-distance dispersal of both cryptic species. Minor sequence variation within clade 2 was partitioned between the islands however, and suggests genetic isolation between clade 2 populations. symposium presentation Lee, Hyang B. 1 *, Kim, Youngjun 2 , Jin, Hui Z. 3 , Lee, Jung J. 3 , Kim, Chang-Jin 3 , Park, Jae Y. 1 , Park, Chae H. 1 and Jung, Hack S. 1 1 Department of Biological Sciences, Seoul National University, Seoul 151-747, Korea, 2 Division of Biotechnology, The Catholic University of Korea, Puchon 420-743, Korea, 3 Korea Research Institute of Bioscience and Biotechnology (KRIBB), Post office Box 115 Yusung, Taejon 305-600, Korea. minervas@snu.ac.kr. A new Hypocrea strain producing harzianum A cytotoxic to tumor cell lines. A new fungal strain producing a trichothecene metabolite, harzianum A, was isolated and its cytotoxicity to tumor cell lines was evaluated. The strain was identified as a new Hypocrea strain based on morphological characteristics and ITS rDNA sequence data. Harzianum A was isolated from wheat bran culture by 50% acetone extraction, silica gel chromatography, Sephadex LH-20 chromatography and HPLC. The chemical structures were determined by ESI- or HRFAB- MS and 1 H and 13 C-NMR analyses. Harzianum A showed cytotoxicity to HT1080 and HeLa cell lines with IC 50 values of 0.65 and 5.07 ug ml -1 , respectively. Harzianum A with a chemical formula of C 23 H 28 O 6 showed moderate to strong cytotoxicity to human cancer cell lines. This is the first report on the production of cytotoxic harzianum A by a new Hypocrea strain. poster Lee, Jin S. 1 *, Sung, Ha Y. 1 , Lim, Young W. 2 and Jung, Hack S. 1 1 Department of Biological Sciences, College of Natural Sciences, Seoul National University, Seoul 151-747, Korea, 2 Department of Wood Science, Faculty of Forestry, University of British Columbia, Vancouver, BC V6T 1Z4, Canada. minervas@snu.ac.kr. Phylogenetic analyses of Perenniporia and Ganoderma based on molecular sequences. Perenniporia s. l., characterized by the ellipsoid to distinctly truncated spores usually with thick walls of variable dextrinoid reaction, is a large heterogeneous group that overlaps with several other generic concepts and makes the classification difficult at present. Phylogenetic relationships of 48 taxa of Perenniporia and related genera were studied by comparing differences among phylogenetic trees inferred from ITS1 rDNA, partial 28S rDNA, and 6-7 regions of RPB2 DNA sequences. It showed that the species of Perenniporia s. l. did not form a monophyletic group and were divided into six subgroups; Abundisporus (A. fuscopurpureus, A. sclerosetosus, Loweporus pubertatis, L. violaceus), Loweporus (L. lividus, L. roseoalbus, L. tephroporus), Perenniporia s. s. (Perenniporia medulla-panis, P. narymica, P. subacida), Perenniporiella (Perenniporiella micropora, P. neofulva), Truncospora (Perenniporia aurantiaca, P. ochroleuca, P. ohiensis), and Vanderbylia (Perenniporia delavayi, P. fraxinea, P. latissima). Besides, another subgroup Ganoderma (G. applanatum, G. meredithiae, G. lucidum, G. resinaceum, Perenniporia robiniophila) with truncate thick-walled spores as a common character was included in Perenniporia s.l. together. poster Lee, Soo Chan* and Shaw, Brian D. Program for the Biology of Filamentous Fungi, Department of Plant Pathology and Microbiology, Texas A&M University, College Station, Texas, 77803, USA. sclee@tamu.edu. The role of protein myristoylation in cell morphogenesis in Aspergillus nidulans. N-myristoylation increases hydrophobicity to allow cytoplasmic proteins to associate with membranes. This modification is mediated by N-myristoyl trasnferase (NMT). In Aspergillus nidulans, the mutation in NMT encoding gene (swoF1) results in abnormal morphogenesis during spore germination and establishment of hyphal growth at restrictive temperature. Six suppressors of swoF1 (ssf) mutants have been identified through UV mutagenesis. Genetic analysis has shown that all six mutations are extragenic to swoF1 and all mutated proteins are downstream of SwoF1. These secondary mutations enable the swoF1 mutant to recover from the loss of cell polarity axis. All ssf mutants have been separated form swoF1 strain by backcross with wild type. Interestingly ssfB, ssfC, and ssfD produced a red pigment, which could be ascoqunoine, which is produced during ascosporogenesis, or norsolorinic acid, a precursor of sterigmatocystin. The distinguishable colonial phenotype of ssf mutants at 42C enables us to clone each gene by complementation. Through the step, ssfD has been found to encode one subunit of the 26S proteasome, which is likely to interact with another proteasome subunit protein, which is predicted to be myristoylated. Subsequent analysis of ssfD is ongoing. The analysis of these mutants is in progress and will be discussed. contributed presentation Continued on following page Inoculum 56(4), November 2005 35
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 and 8: of fungal glucan synthesis. This in
Page 9 and 10: Barrett, Luke G., Thrall, Peter H.,
Page 11 and 12: one new IGS1 RFLP banding pattern f
Page 13 and 14: first contribution on the knowledge
Page 15 and 16: of diversity in the warm climes of
Page 17 and 18: nizing fungi on bay laurel, both ep
Page 19 and 20: munity of several arctic plants in
Page 21 and 22: and subantarctic regions and conduc
Page 23 and 24: the corresponding growth stages in
Page 25 and 26: Maryland 20705, USA, 3 1089 A Stree
Page 27 and 28: growth at 10 o C on potato-dextrose
Page 29 and 30: Johnston, Peter R. Landcare Researc
Page 31 and 32: esults have shown evidence of repro
Page 33: term survival of Aspergillus flavus
Page 37 and 38: attack by spruce bark beetle (Dendr
Page 39 and 40: Matheny, P. Brandon Dept. of Biolog
Page 41 and 42: examine fungal communities associat
Page 43 and 44: data from four protein-coding genes
Page 45 and 46: Novick, Rachel S. Yale University,
Page 47 and 48: substrates in leaf litter prior to
Page 49 and 50: document remediation after flooding
Page 51 and 52: Sakai, Shunsuke 1a , Nishide, Tatsu
Page 53 and 54: strain was also discussed. The addi
Page 55 and 56: Stadler, Marc 1 *, Hellwig, Veronic
Page 57 and 58: on the telial hosts followed by rad
Page 59 and 60: glucosidase activity was higher com
Page 61 and 62: Upadhyay, Srijana, Long, Melissa an
Page 63 and 64: gon State University and downloaded
Page 65 and 66: transmission electron, and epifluor
Page 67 and 68: sequence of Monascus sp. BCRC 38072
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Page 71 and 72: MYCOLOGIST’S BOOKSHELF This issue
Page 73 and 74: MYCOLOGIST’S BOOKSHELF Previously
Page 75 and 76: Ascomycota of Sweden www.umu.se/myc
Page 77 and 78: inoculum The Newsletter of the Myco
Page 80: An Invitation to Join MSA THE MYCOL

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