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IMC7 Main Congress Theme V: CELL BIOLOGY AND PHYSIOLOGY Posters sterile elements occur before the formation of mature basidium. The process of cell senescence and death in B (Aphyllophorales) occur in the direction from the trama of pileus to trama hymenophore and subhymenium. In FB of Agaricales and Gasteromycetes this processes occur in direction, from the base of stipe to its upper part, later they occupied the trama of pileus, trama pf hymenophore and subhymenium. It was supposed that during the morphogenesis of B and FB in Holobasidiomycetes they are early transforming in to the autonomous from mycelium way of morphogenesis. 1186 - Construction of a gene-trap vector, pPTR- EGFP1 and pUsCdelPA2, for Aspergillus oryzae S. Suzuki * & Y. Kashiwagi National Food Research Institute, Kan-nondai 2-1-12 Tukuba Ibaraki 305-8642, Japan. - E-mail: satosuz@nfri.affrc.go.jp A promoter-trap vector that uses GFP as a reporter for Aspergillus oryzae gene expression and a polyA-trap vector using auxotrophic selection marker were constructed for the first time. The promoter-trap vector, pPTR-EGFP1, and polyA-trap vector, pUsCdelPA2 can be used to investigate unknown gene function. These gene-trap vector were integrated into a host genome randomly and transformants were selected by drug resistans or auxotrophic marker gene on the vector. The endogenous promoter activity trapped by the pPTR-EGFP1 vector was used to express the reporter gene egfp. The endogenous polyA addtion signal trapped by the pUsCdelPA2 vector was used to express the auxotrophic marker gene sC on the vector. By the transformation using pPTR-EGFP, approximately 300 drug resistant transformants were obtained, one of which showed the same strong fluorescence as GFP. The expression of egfp gene was confirmed by Northern blot analysis. The flanking genome regions of the integrate site was isolated by plasmid rescue method. Our resuls show that this newly constructed vector, pPTR-EGFP1, can trap endogenous promoter activity. The transformation using pUsCdelPA2 is under continuation now. 1187 - Production of CsmA, a class V chitin synthase with a myosin motor-like domain of Aspergillus nidulans, is regulated in multiple ways N. Takeshita, A. Ohta & H. Horiuchi * Department of Biotechnology, The University of Tokyo, 1- 1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. - E-mail: ahhoriu@mail.ecc.u-tokyo.ac.jp Chitin, a β-1,4-linked homopolymer of Nacetylglucosamine (GlcNAc), is one of the major structural components of the fungal cell wall. Chitin synthases are membrane-bound proteins and have been classified into five groups, classes I to V, on the basis of the structures in 360 Book of Abstracts their conserved region. csmA gene of Aspergillus nidulans encodes a class V chitin synthase with a myosin motor-like domain at its N-terminus. csmA null mutants showed remarkable abnormalities in polarized growth, hyphal wall integrity, and conidiophore development. In this study, to investigate the intracellular behavior of the csmA product (CsmA) and the regulation of its production, we constructed strains that produced CsmA tagged with nine repeats of hemagglutinin A (HA) epitope at its C-terminus, CsmA-HA, instead of CsmA. Western blot analysis with anti-HA antibody proved that the entire coding region of csmA was translated as a single polypeptide with an approximate molecular mass of 210 kDa. CsmA-HA was found in the membrane fraction of the CsmA-HA producing strain. CsmA-HA was produced during vegetative growth, and its amount increased under low osmotic conditions. A discrepancy in the amounts of CsmA-HA protein and csmA transcript as determined by northern analysis under those conditions suggests that the production of CsmA is regulated at both the transcriptional and post-transcriptional levels. 1188 - Media optimization for manganese peroxidase production by response surface methodology in a newly isolated white-rot fungus Trichophyton rubrum LSK-27 C. Tamerler 1* , U.O.S. Seker 1 , K. Li 3 , H. Jung 2 & H. Bermek 1 1 Istanbul Technical University, Molecular Biology and Genetics, Maslak - Istanbul 80626, Turkey. - 2 Sunchon National University, Department of Forest Resources, Sunchon 540-742, Korea. - 3 Oregon State University, Department of Wood Science and Engineering, 102 Richardson Hall, Corvallis, OR97331-5751, U.S.A. - Email: tamerler@itu.edu.tr Response surface methodology (RSM) is a useful technique for media optimization, which is a crucial step for maximizing enzyme production in fungal cultures. In current study, RSM was applied for media optimization in a newly isolated white rot-fungus T. rubrum LSK-27, for manganese peroxidase (MnP) production. Carbon, nitrogen and also phosphorus are known to have significant impact in the production of fungal enzymes. Manganese stimulates MnP production in most fungi. In the first optimization step, glucose (x1), mycological peptone (x 2), MnSO 4 (x 3) and KH 2PO 4 (x 4) were studied to investigate their effect on MnP production by fractional factorial design, 2 4_1 . The first order equation with the regression coefficients were computed to be y=0.7175+0.0625x 1+0.4318x 2-0.1201x 3- 0.0364x 4 with a probability level of 91% for MnP production. Although glucose and KH2PO 4 positively affected the cell growth, they did not increase the enzyme levels. Among the response curves obtained for four media components, the highest response was obtained for MnSO4-peptone pair, which was chosen as main factors for central composite design. Effect of each media ingredient on fungal morphology was also followed. The optimum MnSO 4-peptone concentrations were found for maximum MnP production.
IMC7 Main Congress Theme V: CELL BIOLOGY AND PHYSIOLOGY Posters 1189 - Expression studies of plant genes differentially expressed in leaf and root tissue of arbuscular mycorrhizal (AM) fungal colonised tomato plants J. Taylor & L.A. Harrier * Scottish Agricultural College, Kings Buildings, West Mains Road, Edinburgh, Lothian, Scotland, U.K. - E-mail: l.harrier@ed.sac.ac.uk Arbuscular mycorrhizal (AM) fungi of the unique phylum Glomeromycetes are a multifaceted group of mutualistic symbionts that are common to terrestrial ecosystems. The interaction between AM fungi and plant root systems is of environmental and agronomic importance. Understanding the molecular changes within the host plant upon AM fungal colonisation is a pre-requisite to a greater understanding of the mechanisms underlying the interaction. Differential mRNA display was conducted on leaf tissue of tomato plants colonised and non-colonised by the AM fungus Glomus mosseae and five putative differentially regulated cDNAs were identified. All cDNAs isolated shared high sequence similarity to known plant genes. Semi-quantitative RT-PCR was used to establish gene expression patterns for all five clones within leaf and root tissue of mycorrhizal and non-mycorrhizal colonised tomato plants. Differential regulation was observed for all five cDNAs. Down-regulation within the leaf tissue of mycorrhizal plants was observed for 4 out of the 5 cDNAs with an up-regulation observed only for one. Tissue specific regulation was observed for several cDNAs, with down-regulation observed in mycorrhizal leaf tissue and up-regulation observed within mycorrhizal root tissue as compared to non-mycorrhizal tissue. We thank the Scottish Executive Environmental Rural Affairs Division (SEERAD) for financial support. 1190 - Application of AFLP markers to genetic study of an edible mushroom, Lentinula edodes: strain typing, genetic diversity, and genetic linkage map K. Terashima 1* , T. Matsumoto 2 , E. Hayashi 3 & Y. Fukumasa-Nakai 2 1 Japan Science and Technology Corporation (JST), Honmachi 4-1-8, Kawaguchi, Saitama, 332-0012, Japan. - 2 The Tottori Mycological Institute, Kokoge211, Tottori, 689-1125, Japan. - 3 Forest Tree Breeding Center, Ishi, Juo, Taga, Ibaraki 319-1301, Japan. - E-mail: ktera@hal.ne.jp Lentinula edodes (shiitake) is one of the most popular edible mushrooms in east Asia (Japan, Korea and China). It is desirable to establish efficient breeding systems of this mushroom using molecular markers. In this study, we evaluated the usefulness of amplified fragment length polymorphism (AFLP) as genetic marker in L. edodes. Strain typing and genetic diversity: Six AFLP primer pairs reproducibly detected 179 polymorphic DNA fragments among 15 strains of L. edodes currently used for cultivation in Japan. These markers could differentiate all of the strains. Cluster analysis and principle coordinates analysis based on AFLP data revealed two groups which corresponded to those categorized on their fruiting season types. Genetic linkage map: A medium-dense genetic linkage map of L. edodes was constructed based on 203 AFLP markers and two mating type factors. The segregation of these markers was generated from 95 progeny of a single cross of two distantly related L. edodes strains. Segregation analysis showed that the map consisted of 11 linkage groups, and the total genetic distance of the map was 1956.7cM. The average rate of physical size to genetic distance could be roughly estimated to be less than 18.4 kb/cM, which is low compared to the values obtained for other filamentous fungi. Seventeen of the AFLP markers showed highly distorted segregation ratios (χ 2 values ≥ 6.63; P ≤ 0.01), and many of these were located in LG II (6 markers) and IV (6 markers). 1191 - pH optimization of AM symbiosis with Glomus intraradices and carrot transformed roots in vitro P. Tiwari 1* , U.G. Reddy 1 , A. Prakash 2 & A. Adholeya 1 1 Centre for Mycorrhizal research, Tata Energy Research, Institute, Habitat Centre, Lodi Road, New Delhi-110003, 2 India. - School of Biotechnology, Department of Microbiology, Barkatullah University, Bhopal,Madhya Pradesh, India. - E-mail: pragati@teri.res.in AM fungi Glomus intraradices was introduced on M media with different pH regimes ranging from pH 4 to pH 13 in vitro. The symbiosis was found to express differently over the acid and alkaline range. The investigation was directed to study the difference in expression of AM fungi in terms of inter and intraradical spread, spore formation, mycelial and root biomass along with nutritional uptake in roots. pH 8 was observed to be more supportive to maximum AMF symbiosis, also supporting maximum root biomass. pH 9 supported more vegetative proliferation of the fungus. An increasing trend was observed in mycorrhizal colonization percentage (MCP) from pH 4 to 13. The study is significant in optimizing parameters responsible for optimum symbiosis and exploit its potential in mass inoculum production. 1192 - Comparative symbiotic events of various coal ash amendments in vitro on AM fungi Glomus intraradices and Ri T-DNA transformed carrot roots P. Tiwari 1* , U.G. Reddy 1 , A. Prakash 2 & A. Adholeya 1 1 Centre for mycorrhizal research, Tata Energy Research Insitute, Habitat Centre, Lodi Road, New Delhi, 110003, 2 India. - School of Biotechnology, Department of Microbiology, Barkatullah University, Bhopal, Madhya Pradesh, India. - E-mail: pragati@teri.res.in Book of Abstracts 361
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Page 359: IMC7 Main Congress Theme V: CELL BI
Page 367 and 368: Cavernularia: 356 Cenococcum: 500,
Page 369 and 370: Hyalorbilia: 479 Hyalorhinocladiell
Page 371 and 372: Phlebopus: 828 Pholiota: 304, 515 P
Page 373 and 374: Verrucaria: 616 Verruculina: 963 Ve
Page 375 and 376: Order Index Agaricales: 40, 50, 51,
Page 377 and 378: Bogomolova, E.V.: 1078 Bohar, Gy.:
Page 379 and 380: Forlani, G.: 565 Forsby, A.: 842, 8
Page 381 and 382: Juuti, J.T.: 701, 1126 Kabrick, J.M
Page 383 and 384: Morocko, I.: 859 Moser, J.C.: 426,
Page 385 and 386: Sattayaphisut, W.: 1171 Saunders, E
Page 387 and 388: Vukojevic, J.: 363 Vurro, M.: 116 V
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