Protoplast fusion and genetic recombination in Metarhizium anisopliae |
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| Affiliation: | 1. Department of Biochemical Engineering, Faculty of Science and Technology, University of Debrecen, Egyetem tér 1, H-4032 Debrecen, Hungary;2. Fungal Physiology, Westerdijk Fungal Biodiversity Institute & Fungal Molecular Physiology, Utrecht University, Uppsalalaan 8, 3584 CT Utrecht, the Netherlands;1. School of Food Sciences and Engineering, South China University of Technology, Guangzhou City, Guangdong Province 510640, China;2. Department of Microbiology, Guangzhou Center for Disease Control and Prevention, Qide Road No.2, Guangzhou City, Guangdong 510440, China;3. College of Food Science and Technology, Guangdong Ocean University, Zhanjiang City, Guangdong Province 524088, China;1. School of Forest Resources and Conservation, University of Florida, Gainesville, FL, 32603, USA;2. Department of Entomology and Nematology, University of Florida, Gainesville, FL, 32603, USA;3. Institute of Microbiology, Czech Academy of Sciences, Vídeňská 1083, 142 20 Prague 4, Czech Republic;1. Department of Entomology, Cornell University, Comstock Hall, 129 Garden Avenue, Ithaca, NY 14853, USA;2. USDA Forest Service, Southern Research Station, 200 W. T. Weaver Blvd., Asheville, NC 28804, USA;3. USDA Agricultural Research Service, Robert W. Holley Center for Agriculture & Health, 538 Tower Road, Ithaca, NY 14853, USA;4. Department of Forestry, Wildlife and Fisheries, University of Tennessee, 2506 Jacob Drive, Knoxville, TN 37966, USA;1. Department of Pharmaceutical Botany, Jagiellonian University, Collegium Medicum, Medyczna St 9, Kraków 30-688, Poland;2. Department of Inorganic and Analytical Chemistry, Jagiellonian University, Collegium Medicum, Medyczna St 9, 30-688 Kraków, Poland;3. Faculty of Materials Science and Ceramics, AGH University of Science and Technology, Kraków 30-059, Poland |
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| Abstract: | Using two strains of Metarhizium anisopliae var. minor designated E6 and RJ from different origins, inter-strain crosses were readily performed by orthodox methods from mutants of strain E6 through the parasexual cycle. However, in most cases crosses between mutants of strain RJ or between RJ and E6 mutant strains were not achieved. Protoplast fusion was carried out in an attempt to cross such strains. Protoplasts were obtained after treatment of mycelium with lytic enzymes using 0.7 m KCl as osmotic stabilizer. Regeneration frequency varied from 1.2 to 3.0%. Poly (tethylene glycol) was used for the fusion of protoplasts and fusion frequencies varied from 0.9 to 44.1 in 105 protoplasts according to the cross. Sectors which emerge from fusion products were analysed and recombinants were obtained even from crosses between mutant strains which could not be crossed by hyphal fusion. In this way protoplast fusion proved to be a valuable tool for further studies of genetics and breeding of Metarhizium anisopliae. |
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