Plant regeneration from protoplasts of Triticum aestivum L. cv. Nakasoushu |
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Authors: | Li Hongchao Machii Hiroaki Hagio Takashi Takezaki Akane Hirabayashi Toshio |
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Affiliation: | (1) National Institute of Agrobiological Resources, Kannondai, Tsukuba, Ibaraki 305-8602, Japan;(2) Frorimond Desprez Biotechnology Laboratory, P.O.Box 41, 59242 Cappelle en Pevele, France;(3) National Institute of Sericultural and Entomological Science, Owashi, Tsukuba, Ibaraki 305-8634, Japan;(4) National Institute of Fruit Tree Science, Okitsu, Shimizu, Shizuoka 424-0292, Japan;(5) Shikoku National Agricultural Experiment Station, Senyu-cho, Zentsuji, Kagawa 765-8508, Japan |
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Abstract: | A fast-growing, small, granular, embryogenic callus was selected from primary calli induced from the Japanese wheat cultivar Nakasoushu and the Australian wheat cultivar Bodallin. Regenerable and fine suspension cultures were induced three to six months after liquid culture was initiated and were characterized by dense cytoplasm and active division. These suspension cultures routinely provided high yields of protoplasts with about 90% viability when incubated in a modified KMP (Kao and Michayluk, 1975) medium containing 1 mg l-1 2,4-D (2,4-dichlorophenoxyacetic acid), and 1 mg l-1 zeatin. Nakasoushu and Bodallin protoplasts divided at frequencies of 8.6% and 11.1%, respectively, in agarose-solidified media. When Nakasoushu protoplasts were cultured with effective nurse cells of sorghum and wheat, protoplast division increased to 16.9% and 12.6%, respectively. Plating efficiencies varied from 0.03% to 2.5%. After subculture, protocolonies yielded embryogenic calli and somatic embryos, from which green plants were eventually regenerated. Whole plants obtained from Nakasoushu protoplasts were fertile, demonstrating the first report of Japanese cultivars in wheat protoplast cultures. This revised version was published online in June 2006 with corrections to the Cover Date. |
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Keywords: | plant regeneration protoplast suspension culture wheat |
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