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IMC7 Main Congress Theme III: PATHOGENS AND NUISANCES, FOOD AND MEDICINE Posters including number of septa and size of conidia, among Indonesian isolates of C. heterostrophus existed, an indication that Indonesian C. heterostrophus might have originated from different ancestors. In contrast, pathogenicity of each isolate measured by disease incidence on several corn varieties was not significantly different. Disease severity of the corn varieties inoculated with each isolate was also relatively the same. The results imply that the corn varieties tested possess potential risks of southern corn leaf blight outbreaks. 824 - Diagnosis of cryptic microbes based on ITS2 rRNA secondary structure information A. Gargas 1* & P.T. DePriest 2 1 Dept. of Botany, University of Wisconsin, Madison, 132 Birge Hall, 430 Lincoln Dr, Madison, WI 53706, U.S.A. - 2 Dept. of Botany, National Museum of Natural History, Smithsonian Institution, Washington, DC 20560, U.S.A. - E-mail: agargas@facstaff.wisc.edu Conventional techniques of isolating and identifying fungi or other cryptic microbes are time-consuming and expensive, limiting their utility for rapid and widespread surveys. We invented a novel method for detection and identification of microbes including those never before isolated or identified. This approach relies on analyses of the internal transcribed spacer 2 (ITS2) rDNA sequences - highly variable regions flanked by the 5.8S and 28S ribosomal subunits within nuclear DNA. After determining secondary structures of ITS2 rRNAs we identify nucleotide motifs between 20 and 50 nt in length within each primary sequence. These signature sequences, readily defined and present in high copy number, allow taxonomic diagnoses of known and unknown microbes from various substrates including agricultural, medical and ecological samples. Such sequences are readily adapted for use with probing techniques including PCR, microarray assays or other molecular detection methods. 825 - First report of Coniothyrium stem canker on E. camaldulensis in Ethiopia A. Gezahgne * , J. Roux & M.J. Wingfield Department of Plant Pathology and Microbiology, Tree Pathology Co-operative Programme (TPCP), Forestry and Agricultural Biotechnology Institute (FABI), University of Pretoria, Pretoria 0002, South Africa. - E-mail: alemu@fabi.up.ac.za During a survey of Eucalyptus diseases in Ethiopia, a serious stem canker disease was discovered on E. camaldulensis trees at several localities in the South and South Western parts of the country. The disease is characterised by the presence of discrete necrotic lesions, stem cankers, cracking of the stems, production of kino pockets in the wood, as well as malformation of stems. 248 Book of Abstracts These symptoms are similar to those caused by Coniothyrium zuluense in South Africa. The aim of this study was to positively identify the causal agent of the disease in Ethiopia. This was achieved by sequencing the ITS region of the rRNA operon for a representative set of isolates. Sequences for the Ethiopian isolates were compared with those from authenticated isolates collected in South Africa, Thailand and Mexico. Based on these data, the Ethiopian isolates were shown to group more closely with those from South Africa, than with those from other areas. This study represents the first report of C. zuluense and its associated disease in Ethiopia. Currently the disease is causing considerable losses in yield and quality of timber and it also impacts negatively on the lives of subsistence farmers. We are currently studying additional isolates of the fungus and utilising sequences of different genes to study the phylogenetic relationships between C. zuluense, from a wide range of countries. 826 - Enzyme mediated infection of host Pinus short lateral roots by Tricholoma matsutake W. Gill 1 , K. Suzuki 2 , M. Brown 3 & C. Mohammed 4* 1 Tasmanian Institute of Agricultural Research, 13 St Johns Ave, New Town, Tasmania 7008, Australia. - 2 Laboratory of Forest Botany, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan. - 3 Huon Valley Mushrooms, Main Road, Glen Huon, Tasmania 7109, Australia. - 4 School of Agricultural Science, University of Tasmania, GPO Box 252-54, Tasmania 7001, Australia. - E-mail: caro.mohammed@utas.edu.au Tricholoma matsutake (Ito et Imai) Sing. (Matsutake) is an edible mycorrhizal basidiomycete highly prized in Japan for its culinary and medicinal properties and historical social significance. However, the natural supply of Matsutake in Japan is in decline and domestic demand exceeds supply. Consequently, the development of a viable commercial artificial cultivation system for this mushroom is of prime concern. In both natural conditions and artificial culture, Matsutake forms morphologically atypical ectomycorrhizas on host Pinus short lateral roots. Furthermore, aspects of the mushroom's nutrition are unusual and it is this atypical interaction which must be understood in order to devise an artificial cultivation system. Here, we present evidence of host infection and the parallel ability of T. matsutake to form a range of plant cell wall degrading hydrolytic enzymes in artificial culture. The hydrolytic enzymes tested were detected over a range of activities but the majority peaked after 2 or 3 weeks of incubation. Conversely, xylosidase and glucosidase enzymes were detected in comparatively high activities which did not peak but rather increased in activity over the duration of the assay. The significance of the hydrolytic enzymes produced in terms of Matsutake ectomycorrhizal establishment, development and observed morphology is discussed.
IMC7 Main Congress Theme III: PATHOGENS AND NUISANCES, FOOD AND MEDICINE Posters 827 - Molecular probe development for tree root associated Rhizoctonia sp. and Suillus bovinus and detection by Southern dot blot and liquid hybridization H. Grönberg 1* , L. Paulin 2 & R. Sen 1 1 Department of Biosciences, University of Helsinki, Viikinkaari 9, P.O.POX 56, Helsinki- 00014, Finland. - 2 Institute of Biotechnology, University of Helsinki, Viikinkaari 9, P.O.POX 56, Helsinki- 00014, Finland. - Email: henrietta.gronberg@helsinki.fi Specific rDNA ITS (internal transcribed spacer) targeted probes for detection of different conifer root associated Rhizoctonia strains and Suillus bovinus were developed by comparing two different Southern (dot blot and liquid) hybridization methods. Probes (124-151bp) specific to six different Rhizoctonia strains (uninucleate strain 263, binucleate strains 268, AV-2, 251, 266, and 269 and multinucleate R. solani) and one specific to Suillus bovinus and a positive control probe were developed using pure culture DNA. Shorter (20-25 bp) oligonucleotide probes specific to strains 263, 268 and Suillus bovinus were developed in order to obtain more specificity and for used in liquid hybridizations. Total fungal DNA was used as a target with longer DIG (digoxigenin) labeled probes, but in biotin labeled oligonucleotide probes the target DNA was PCR amplified ITS (ITS1, 5.8S, ITS2). The optimal hybridization temperatures obtained from dot blots also gave the strongest signals in liquid hybridization protocol, although low temperature washing (30-35 °C) ocassionally affected to the specificity. We found the liquid hybridization protocol more useful when probing unknown target DNA, as it resolves targets using combined probe hybridization and ITS length polymorphism by resolving fragments differing by 5 bp in a sequencing gel (ALF express). 828 - Ethnomycological knowledge and ecology of Phlebopus sudanicus in Burkina Faso K.M.L Guissou * , Ph. Sankara & S. Guinko Université de Ouagadougou, UFR/SVT, Département de Biologie et Physiologie Végétales, 03 BP 7021 Ouagadougou 03, Burkina Faso. - E-mail: laure_guissou@univ-ouaga.bf The use and the ecology of Phlebopus cf sudanicus is examined in this study undertaken in the Reserve of Hippopotamus pound of Bala (Burkina Faso) and its vicinities. Fixed questions regarding ethnomycological knowledge were submitted to 180 native informants from 6 villages. Collecting field works toward the forest were organised to recognise the mean habitats of this fungus. It appears that a total of 98% of the informants recognise this species as being edible. Two kinds of habitats of Phlebopus sudanicus have been found: under Piliostigma thonningii (Caesalpiniaceae) and Mitragina inermis (Rubiaceae) during the fieldwork. 829 - Genetic diversity of fungi associated with seeds of tropical forest trees (Podocarpus falcatus and Prunus africana) A. Gure * , K. Wahlström & J. Stenlid Department of Forest Mycology & Pathology, Swedish University of Agricultural Sciences, Box 7026, 750 07 Ulls vag 26A, Uppsala, Sweden. - E-mail: Abdella.Gure@mykopat.slu.se The association and interactions between plants and fungi have long been observed and studied. However, the knowledge about the diversity of the fungal flora and the nature of their interactions with their hosts has been very scarce in the area of tropical forest tree seeds. In this study, matured fruits of Podocarpus falcatus and Prunus africana were collected directly from the crowns and from the ground just under the mother trees in March 2000 and May - June 2001. Seed collection was carried out at different forest sites in Ethiopia. Fungi were isolated from fresh fruits, and from seeds with and without seed coat. In all cases, surface sterilised and unsterilised seeds were plated on standard growth media. All in all 150 fungal isolates belonging to 25 genera have been identified by morphological and molecular methods (internal transcribed spacer- ITS sequence analysis) at least to the genus level. Most of the isolates were ascomycetes and a few were basidiomycetes. Some of these fungi are putative seedborne pathogens that will be studied further. 830 - Biocontrol of the Canada thistle (Cirsium arvense) with fungal pathogens S. Guske 1 , B. Schulz 2* & C. Boyle 2 1 Biologische Bundesanstalt, Messeweg, D-38104 Braunschweig, Germany. - 2 Institut für Mikrobiologie, Technische Universität Braunschweig, Spielmannstr. 7, D- 38106 Braunschweig, Germany. - E-mail: b.schulz@tubs.de In order to develop a mycoherbicide for biocontrol of Cirsium arvense (L.) Scop. (Canada thistle), fungal pathogens were tested, both singly and in combination, in semi-field pot experiments in the course of two vegetation periods. Isolates were selected from fungi previously isolated both as pathogens and endophytes from C. arvense and subsequently tested for their virulence. The fungi inoculated singly were Phoma destructiva, Phoma hedericola, Phoma nebulosa, M. sterila isolate 1.4a-4 and the rust, Puccinia punctiformis. Disease severity was evaluated according to disease symptoms (degree of chloroses, necroses, macerations) and various parameters of growth and development, i.e. length and death rates of main and secondary shoots, inflorescences, seed production and germination, wet and dry weights of shoots and roots after harvest. With the exception of Puccinia punctiformis (local infections) all of the isolates applied singly, negatively influenced all measured parameters, those of Book of Abstracts 249
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Cavernularia: 356 Cenococcum: 500,
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Phlebopus: 828 Pholiota: 304, 515 P
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Verrucaria: 616 Verruculina: 963 Ve
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Order Index Agaricales: 40, 50, 51,
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Bogomolova, E.V.: 1078 Bohar, Gy.:
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Vukojevic, J.: 363 Vurro, M.: 116 V
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