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Agrobacterium-mediated transformation of Orychopheagmus violaceus and regeneration of transgenic plants

Zhou Jiming, Wei Zhiming, Xu Zhihong, Liu Shigui, Luo Peng

National Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology, The Chinese Academy of Sciences. Shanghai 200032； China；Department of Biology, Sichuan University.　Chengdu 610064, Sichuan

Abstract

Excised hypocotyls and cotyledons of Orychophragmus violaceus were used as explants for tissue culture. They were cultured on MS medium supplemented with BA 2 mg/L or with BA 3 mg/L and NAA 0.2 mg/L.Regenerated buds were excised when they were 2cm long, and transferred onto 1/2MS medium with IBA 0. 03 mg/L, then whole　plantlets were regenerated at a frequency of 100%. Genetic transformation of O. violaceus was studied by coculture with Agrobacterium tumefaciens strain A208se(pTiT37,pROA93),After 2-3d of coculture the hypocotyls and cotyledens were transferred onto selection medium containing kanamycin(Km) 25　mg/L and 6-aminopenicillanic acia(Ap) 250　mg/L.8 weeks later, shoots emerged, and were excised and transferred onto the rooting medium containing AP 25　mg/L and Cephazoline 100 mg/L,and roots were formed within 4-5 weeks.When the whole plants were transplanted into pots they grew well. The frequency of plant regeneration of hypocotyls was about 30%, and that of cotyledens was about 51%. The regenerated plants showed high enzymatic activities of #beta# glucuronidase and neomycine phosphotransferase II. Southern blot analysis confirmed that NPTII gene had been stably integrated into the chomosomal genome of O.violaceus.The transformation frequency of hypocotyls was 10%, and that of cotyledens was 5.5%.This is the first report about the regeneration of transgenic plants of O.violaceus.

Key words: Agrobacterium tumefaciens;Orychopheagmus violaceus;Hypocotyl;Cotyleden;Transgenic plants

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Citing This Article：

Zhou Jiming, Wei Zhiming, Xu Zhihong, Liu Shigui, Luo Peng. Agrobacterium-mediated transformation of Orychopheagmus violaceus and regeneration of transgenic plants. Acta Physiol Sin 1997; 49 (1): (in Chinese with English abstract).