Bt转基因抗虫甘蓝的研制

Study on Bt transgenic cabbage with insect resistance

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.