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作 者:李贤[1] 姚泉洪[1] 彭日荷[1] 熊爱生[1] 薛永[1] 金晓芬[1]
机构地区:[1]上海市农业科学院生物技术研究所上海市农业遗传育种重点实验室,上海201106
出 处:《上海农业学报》2008年第3期16-20,共5页Acta Agriculturae Shanghai
基 金:上海市科委基础研究重大项目(06DZ19103);上海市科技兴农重点攻关项目[沪农科攻字(2006-2-1)];上海市科委启明星跟踪项目(08QH14021)资助
摘 要:以子叶柄为外植体。通过根癌农杆菌介导法将苏云金芽孢杆菌杀虫蛋白基因CryIA(c)加上叶绿体转运肽RBCSsig.导人结球甘蓝栽培品种"夏光"的母本"103"中,共获得48株卡那霉素抗性植株。经GUS染色和PCR扩增检测,选出5株阳性植株进行连续2代自交授粉结合GUS染色检测。获得了5个CryIA(c)基因纯合株系。RT-PCR证明CryIA(c)基因在5个株系中高表达。离体叶片抗虫试验表明,转基因植株抗性较对照显著提高。大田种植试验发现,转基因株系在不施用杀虫剂的条件下依然可以正常生长直至结球,而对照株系遭受虫害严重。The cotyledon petioles of cabbage (Brassica oleracea var. capitata )were used as explants, and through Agrobacterium tumefaciens-mediated transduction, the insecticidal protein gene CrylA (c)of Bacillus thuringiensis (Bt) and the transit peptide RBCSsig. of chloroplast were introduced into the female parent"103"of cabbage cultivar"Xiaguang", thus producing a total of 48 plants with kanamycin resistance. By GUS assay and PCR detection, five positive plants were selected and their self-pollination was carried out continuously for two generations. The RT-PCR result showed that the CryIA(c) gene was highly expressed in the obtained 5 homozygous CryIA (c) lines. The test for the insect resistance of excised leaves showed that the insect resistance of CryIA (c) transgenic cabbage was significantly greater than that of a control. The field experiment showed that without applying insecticides the transgenic plants could grow normally till heading but the control plants yielded less for heavy damage by insects.
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