根癌农杆菌介导_em_BDN1_em_+基因转化马铃薯的研究.pdfVIP

中国蔬菜　2013（6）：37-43 CHINA VEGETABLES 根癌农杆菌介导BDN1 基因转化马铃薯的 研究 1 1* 1 1 2 潘映雪 　张丽莉 　石　瑛 　卢翠华 　林忠平 1 2 （ 东北农业大学农学院，黑龙江哈尔滨 150030； 北京大学生命科学学院，北京 100 871） 摘　要： 以马铃薯品种东农 303 微型薯为试验材料，通过根癌农杆菌介导法，将来自耐旱植物厚 叶旋 蒴苣苔的脱水蛋白基因 BDN1 导入马铃 薯，并对其遗传转化的影响因素进行了研究。获得了抗性再生植株 35 株 ；PCR 检测 27 株呈阳性；Southern-blot 检测 4 株呈阳性，表明 BDN1 基因已整合到马铃薯基因组中； 半定量 RT-PCR 检测结果表明 BDN1 基因在马铃薯抗性植株中表 达。通过对相对含水量、脯氨酸、丙二醛 含量和 SOD 活性测定，证明 4 个转基因株系对干旱胁迫的抗性显著高于对照植株。 关键词：马铃薯；农杆菌介导；BDN1 基因；抗旱；遗传转化 中图分类号：S532　　文献标识码：A　　 文章编号： 1000-6346（2013）06-0037-07 Studies on Potato Transformation of BDN1 Gene Mediated by Agrobacterium tumefaciens 1 1* 1 1 2 PAN Ying-xue ，ZHANG Li-li ，SHI Ying ，LU Cui-hua ，LIN Zhong-ping 1College of Agronomy Northeast Agricultural University Harbin 150030 Heilongjiang, China 2 College of Life （ ， ， ， ； Science，Peking University ，Beijing 100871 ，China ） Abstract ：The BDN1 gene in coding area from drought-tolerant Boea crassifolia Hemsl was transferred into potato（Solanum tuberosum L.） cultivars‘ Dongnong 303’ through agro-bacterium- mediated transformation. The factors influencing its genetic transformation were studied. Thirty-five transgenic plants with kanamycin resistance were obtained. Specific primers were used to amplify the full length of the BDN1 gene from total genomic DNA of the transgenic plants. Among them, 27 plants were positive by PCR test and 4 plants were positive by Southern-blot test. The result of semi-quantitative RT-PCR test showed that BDN1 gene was inserted into the genome of the transgenic plants and can be transcribed normally. The physiological index showed the transgenic potato plants were signific