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IMC7 Main Congress Theme III: PATHOGENS AND NUISANCES, FOOD AND MEDICINE Posters 864 - Simultaneous detection of GFP- and GUS-marked fungi of different formae speciales of Fusarium oxysporum on plant roots T. Nonomura * , Y. Matsuda & H. Toyoda Laboratory of Plant Pathology and Biotechnology, Faculty of Agriculture, Kinki University, 3327-204 Nakamachi Nara 631-8505, Japan. - E-mail: nonomura@nara.kindai.ac.jp In our attempt to visualize infection behavior of the fungal wilt pathogens inoculated onto plant roots, the fungi were genetically marked with two reporter genes. F. oxysporum f. sp. lycopersici (FOL) and F. o. f. sp. melonis (FOM) were transformed with the green fluorescence protein gene (GFP) and the β-glucuronidase gene (GUS), respectively. In the present study, we focused mainly on the attachment and subsequent hyphal elongation by microconidia inoculated onto roots of tomato and melon seedling. In addition, we attempted to directly distinguish different formae speciales of F. oxysporum onto the same plant roots by expression of different marker genes. Microconidia of GFP-marked FOL (KFOL-001) and GUS-marked FOM (KFOM-002) were inoculated onto roots of cotyledonal seedlings, and inoculated roots were first observed under a fluorescence microscope to detect KFOL-001 and then stained with X-gluc (substrate for GUS assay) to detect KFOM-002 under a light microscope. Consequently, both transformed pathogens could be clearly distinguished at the same site of inoculation. These results suggest that dual transformation of F. oxysporum is useful for analyzing behavior of nonpathogenic F. oxysporum challengeinoculated with pathogenic F. oxysporum. 865 - Biscogniauxia and Daldinia; latent pathogens of deciduous trees L.K. Nugent * , G.P. Sharples & A.J.S. Whalley School of Biomoleculr Sciences, Liverpool John Moores University, Byrom Street, Liverpool L3 3AF, U.K. - E-mail: beslnuge@livjm.ac.uk Biscogniauxia Kuntze and Daldinia Ces. and De Not. are two wood inhabiting xylariaceous genera. Biscogniauxia species are frequently linked with canker diseases in stressed hosts e.g. B. mediterranea causes coal canker in Quercus suber (cork oak) and B. nummularia canker in Fagus (beech) while D. concentrica (Bolt. ex. Fr.) causes calico wood in Fraxinus (ash). Studies on ascospore germination and development of the anamorphs in culture in response to host extracts is presented. Biscogniauxia nummularia and Daldinia concentrica. have been isolated from their respective host leaves and branches and there are frequency of isolation maybe linked to ascospore production. The presence of Daldinia in leaves and in wood has been investigated microscopically, chemically and by molecular techniques, in addition to traditional isolation techniques following surface sterilisation. The 260 Book of Abstracts presence of latent pathogens and their relationship to stress of the host is presented. Experimentation on conditions leading to the latent invasion and subsequent development of teliomorphs are being undertaken in both field and laboratory. 866 - Establishment of the first Karnal Bunt testing laboratory in South Africa O.M. O'Brien, I.H. Rong * & E.J. van der Linde ARC-Plant Protection Research Institute, Private Bag X134, Pretoria 0001, South Africa. The fungal wheat disease Tilletia indica, commonly known as Karnal Bunt (KB), was detected in a limited area of one of South Africa's wheat producing regions during 1999. Aimed at managing T. indica, a national survey to determine the occurrence of the disease was initiated by the Directorate Plant, Health and Quality, National Department of Agriculture. To this end, the Mycology Unit, ARC-Plant Protection Research Institute, Pretoria was tasked with setting up a laboratory for the analyses of seed and grain samples. Due consideration was given to the geographical distance of the laboratory from the main wheat producing areas of the country. The KB protocol, as recommended by the USDA/APHIS, was followed with some adaptations. Analyses were conducted for two consecutive years, providing valuable experience in managing a quarantine analytical facility of this nature. Protocols and procedures representing different phases of the process were devised for each workstation. These phases included: reception and registering of samples, sub-sampling, washing and sieving, centrifuging and preparing of microscope slides, detection of T. indica, data processing and reporting, and waste management. Laboratory procedures, problems encountered and the development of novel techniques, as well as the management and maintenance of the quarantine facility, are discussed. 867 - Aflatoxins in the weaning food of Kenyan children S.A. Okoth Botany Department, University of Nairobi, P. O. Box 30197, Nairobi, Kenya. - E-mail: dorisokoth@yahoo.com Cereal grains run a high risk of mycotoxin contamination yet they form the basis of gruels used in weaning children in Kenya. To these grains (maize, sorghum, millet) supplements such as cassava, groundnuts, beans, and fish are added and ground together, depending on the means and education of the parents. Among the mycotoxins, aflatoxins have been implicated in human diseases, including kwashiorkor. Sampling for aflatoxin contamination was done in Kisumu District, Kenya, an area with high relative humidity and temperatures, high incidence of kwashiorkor and the highest prevalence of absolute poverty, 63%, in the country. A total of 180
IMC7 Main Congress Theme III: PATHOGENS AND NUISANCES, FOOD AND MEDICINE Posters samples of the weaning flour were collected from Winam Division of Kisumu District, Kenya. These samples were analysed for aflatoxins in order to find out whether there exists a relationship between the high incidence of kwashiorkor and aflatoxin contamination. The samples were analysed by thin layer chromatography. Fifty two samples (29%) were positive for aflatoxins (concentration range 2-82 µg/kg), some exceeding the advisory limit. This indicates that aflatoxin is likely to present a major health hazard to weaned children in this part of Kenya. 868 - Prevention of ochratoxin A in cereals - an EU project within Quality of Life and Management of Living Resources M. Olsen 1* , N. Jonsson 2 , N. Magan 3 , J. Banks 4 , C. Fanelli 5 , A. Rizzo 6 , A. Haikara 7 , A. Dobson 8 , J.C. Frisvad 9 , S. Holmes 10 , E. Pajunen 11 , S.-J. Persson 12 & A. Jonsson 13 1 National Food Administration, PO Box 622, SE-751 26 Uppsala, Sweden. - 2 Swedish Institute of Agricultural and Environmental Engineering, P.O. Box 7033, SE-750 07, Sweden. - 3 Cranfield Biotechnology Centre, Cranfield University, Barton Road, Silsoe, Bedfordshire MK45 4DT, U.K. - 4 Central Science Laboratory, Sand Hutton, York YO41 1LZ, U.K. - 5 Laboratorio di Micologia, Università "La Sapienza", Largo Cristina di Svezia 24, I-00165 Roma, Italy. - 6 National Veterinary and Food Res. Inst., P.O. Box 45, FIN-00581, Helsinki, Finland. - 7 VTT Biotechnology, P.O. Box 1500, FIN-02044 Espoo, Finland. - 8 Microbiology Department, University College Cork, National University of Ireland, Cork, Ireland. - 9 BioCentrum-DTU, Technical University of Denmark, Building 221, Søltofts Plads, DK-2800 Kgs. Lyngby, Denmark. - 10 ADGEN Ltd, Nellies Gate, Auchincruive, Ayr KA6 5HW, U.K. - 11 Oy Panimolaboratorio- Bryggerilaboratorium AB, P.O. Box 16, FIN-02150 Espoo, Finland. - 12 Akron Maskiner, SE-531 04 Järpås, Sweden. - 13 Svenska Lantmännen, Östra hamnen, SE-531 87 Lidköping, Sweden. - E-mail: mool@slv.se Ochratoxin A (OTA) is a mycotoxin of considerable concern for human health and is classified as a possible human carcinogen. Cereals normally account for 50-80% of average consumer intake of OTA. Consequently, prevention of OTA formation by specific moulds in cereals would have a significant impact on levels of human exposure. The aim of this multidisciplinary project is to identify the key elements in an effective HACCP programme for OTA in cereals, and provide tools for preventative and control procedures. The project includes the whole food chain from field to the final processed product. The project objectives are divided into 4 different tasks, all important steps in a HACCP managing programme for ochratoxin A in cereals. Task 1: Identification of the critical control points (CCP). The ochratoxin producing species are being identified and characterised in order to identify the sources of contamination. Task 2: Establishment of critical limits for the CCP's. New knowledge concerning the microbial ecology of the ochratoxin producing species is provided and critical limits are being specified. Task 3: Developing rapid monitoring methods. Rapid detection methods for ochratoxin A and for the producing fungi are being developed. Task 4: Establish corrective actions. Reduction during cereal processing (milling etc) and during malting and brewing, is being investigated. 869 - Biological efficacy of Trichoderma harzianum against wilt disease of sugarcane W. Ouvanich Plant Pathology and Microbiology Division, Department of Agriculture, Paholyothin Road, Jatujak, Bangkok 10900, Thailand. - E-mail: wantanee@doa.go.th Biocontrol efficacy of Trichoderma harzianum isolate Th1 and Th2, against sugarcane wilt disease caused by Fusarium subglutinans and Cephalosporium sacchari were determined. Result from pot experiment show that both isolates of T. harzianum were able to significantly reduced wilt pathogen population in soil and wilt infection percentage of sugarcane roots. Their effect on wilt disease has been further studied in field experiment. Each isolate of T. harzianum was grown on sorghum seeds and then mixed the infested seeds with rice polish and manure. The mixture of each isolate of T. harzianum was applied to wilt pathogens infested field soils. Population of wilt pathogens in soil, percentage of infected root and yield of sugarcane were determined in all treatments. Population of wilt pathogens in infested field soil were significantly reduced by both T. harzianum Th1 (2.1 × 103 cfu/g soil ) and T. harzianum Th2 (2.8 × 103 cfu/g soil), as compare with control (5.5 × 103 cfu/g soil). Percentage of wilt infected root only was reduced by T. harzianum Th2 (20.4%). Significant difference in percentage of wilt infected root was not observed between T. harzianum Th1 (25.3%) and control (26.3%) treatments. Yield of sugarcane in T. harzianum Th2 treatment was also higher than in T. harzianum Th1 and control treatments. T. harzianum isolate Th2 has shown to be the effective biocontrol agent of wilt disease of sugarcane. 870 - Natural flavonols fungitoxic to formae speciales of Fusarium oxysporum pathogenic to some ornamentals C. Pasini * , P. Curir, G. Brofiga & F. D'Aquila Istituto Sperimentale per la Floricoltura, Corso Inglesi, 508 - 18038 Sanremo (Im), Italy. - E-mail: difesa@istflori.it Three constitutive flavonols from as many carnation (Dianthus caryophyllus) cultivars, resistant to Fusarium oxysporum f. sp. dianthi pathotype 2, have been assayed, alone or in combination, against the six following formae speciales of F. oxysporum affecting ornamental plants: f. sp.dianthi pathotypes 2 and 8, f. sp. asparagi, f. sp. ranunculi, f. sp.cyclaminis, f. sp. lilii. The fungitoxic effect was tested both as mycelial growth and as spore germination inhibition. The results show that some Book of Abstracts 261
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Cavernularia: 356 Cenococcum: 500,
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Order Index Agaricales: 40, 50, 51,
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Vukojevic, J.: 363 Vurro, M.: 116 V
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