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1-磷酸甘露醇脱氢酶基因转化水稻的研究

Transformation of Rice with mtlD Gene

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【作者】 王慧中刘俊君卢德赵赵艳颜美仙钱前彭学贤陈受宜黄大年

【Author】 WANG Hui zhong 1,2 , LIU Jun jun 3, LU De zhao 2, ZHAO Yan 4, YAN Mei xian 2, QIAN Qian 2, PENG Xue xian 3, CHEN Shou yi 3, HUANG Da nian 2 ( 1 Key Laboratory of Biochemistry and Molecular Biology, Hangzhou Normal College, Hangzhou 310036, China; 2China National Rice Research Institute, Hangzhou 310006, China; 3Laboratory of Plant Biotechnology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Bejing 100101, China; 4Hangzhou Institute of Commerce, Hangzhou 310035, China)

【机构】 杭州师范学院生物化学与分子生物学重点实验室中国科学院遗传与发育生物学研究所植物生物技术开放实验室中国水稻研究所杭州商学院中国水稻研究所 浙江杭州310036浙江杭州310006北京100101浙江杭州310035浙江杭州310006

【摘要】 PCR和 Southern blotting检测表明 ,来自大肠杆菌的 mtl D基因已通过农杆菌介导整合进水稻基因组。mtl D基因在 T1代出现分离 ,T2 代出现纯系。在 0 .75% Na Cl胁迫下 ,7个转基因 T3 代株系都能检测到 mtl D酶活性 ,与对照相比细胞膜的相对电导率和大分子渗漏值明显降低。部分转基因株系能在 1 .0 % Na Cl浓度下正常生长 ,而对照在 0 .5% Na Cl浓度下已不能存活。通过有性杂交途径实现了 mtl D和 gut D两个基因的聚合 ,部分杂交后代株系能在 1 .2 5% Na Cl胁迫下正常生长结实。

【Abstract】 PCR and Southern blot analysis indicated that mtlD gene had been integrated into the rice genome mediated by Agrobacterium tumefaciens LBA4404 (pBIM). mtlD gene segragated in T 1 generation and pure transgenic lines appeared in T 2 generation. The expression of the mtlD gene in transgenic plants was demonstrated by enzymatic activity assay. Under 0 75% NaCl stress, the relative electronic conductivity and membrane permeability were detected in T 3 transgenic plants of 7 different lines. Results showed that the damage to membrane structure of the transgenic plant was lower than that of their controls. Some transgenic plant lines could grow normally under 1.0% NaCl stress whereas the controls could not grow and died after 2 weeks under the same environment. Through sexual hybridization, rice plants with mtlD gene and gutD gene were got, some of them could grow normally under the stress of 1.25% NaCl.

【基金】 浙江省自然科学基金资助项目(3982 97;30 2 0 61 );国家自然科学基金资助项目(39970 4 0 9);中国水稻科学发展基金资助项目 (0 0 0 32 39)
  • 【文献出处】 中国水稻科学 ,Chinese Journal of Rice Science , 编辑部邮箱 ,2003年01期
  • 【分类号】S511
  • 【被引频次】32
  • 【下载频次】225
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