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Preparation and Regeneration of Protoplasts from Axenic Mycelia Derived from the Wheat Stem Rust Fungus. D. Huang, Peoples Republic of China visiting scholar and scientist, Boyce Thompson Institute, Cornell University, Tower Road, Ithaca, NY 14853, Present address: Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Malianwa, Beijing, PRC; R. C. Staples(2), W. R. Bushnell(3), and D. J. Maclean(4). (2)Peoples Republic of China visiting scholar and scientist, Boyce Thompson Institute, Cornell University, Tower Road, Ithaca, NY 14853; (3)(4)Research plant physiologists, Cereal Rust Laboratory, Agricultural Research Service, U.S. Department of Agriculture, University of Minnesota, St. Paul 55108, (4)Present address: Department of Biochemistry, University of Queensland, St. Lucia, Qld. 4067, Australia. Phytopathology 80:81-84. Accepted for publication 1 August 1989. Copyright 1990 The American Phytopathological Society. DOI: 10.1094/Phyto-80-81.

Protoplasts were prepared from axenic suspension cultures of the wheat stem rust fungus Puccinia graminis f. sp. tritici, race 126 Anz-5, 6, 7, 11. Protoplasts were released in 3 hr from 3- to 7-day-old mycelia with a mixture of Novozym 234 and cellulase (5 mg each/ml) osmotically stabilized by 0.8 M MgSO₄. Protoplast regeneration was detected microscopically within 10 hr after incubation in a nutrient medium containing 0.5 M sorbitol, and colonies usually were visible after a further 3 days of growth. The regeneration frequency ranged from 19 to 39%.