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《Acta Biologiae Experimentalis Sinica》 1999-01
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PRELIMINARY STUDIES ON TISSUE CULTURE AND AGROBACTERIUM-MEDICATED TRANSFORMATION OF BRASSICA CAMPESTRIS SSP. CHINENSIS

WANG Ling Jian NI Di An WANG Guang Yuan XIA Zhen Ao XU Zheng Kai (Shanghai Institute of Plant Physiology, Chinese Academy of Sciences, Shanghai 200032)  
The hypocotyls and cotyledons of the asepet-ic seedling of Brassica campestris ssp. chinensis L cv. Pudongaijiecai) were used as explants for tissue culture. Adventitious buds were differentiated on modified MS medium supplemented with TDZ 1 - 2 mg/L, NAA 0. 2 - 1 mg/L and AgNO, 7. 5 mg/L. The percentage of explants which formed buds of cotyledons was about 56%, and that of hypocotyls was about 37 %. When the regenerated explants were transferred onto MS medium with 2ip 5 mg/L and NAA 0.1 mg/L for two weeks, whole plantlets were obtained by culturing the regenerated shoots on 1/2 MS medium with NAA 0.1 mg/L. Agrobacterium tumefaciens strain ( LBA 4404/PBI 121) carrying the GUS gene and Npt Ⅱ gene was used for transformation. After 2 days of coculture, the hypocotyls and cotyledons were transferred onto regenerated medium containing CP 300 mg/L for bud formation. After 4 - 5 weeks, the differentiated buds were transferred onto selection medium with CP 200 mg/L and Km 10 mg/L for 1 month, then the green shoots were transferred onto the rooting medium containing Cef 100 mg/L and Km 20 mg/L. 4 - 5 weeks later, plantlets with Km resistance were obtained and some of them showed higher enzymatic activities of β-glucuronidase than control
【Fund】: 中国科学院重大项目;; 植物分子遗传国家重点实验室的资助
【CateGory Index】: S636;
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