抗虫相关基因KTI 对青花菜的转化及其对小菜蛾抗性的分析

 以青花菜（Brassica oleracea L. ssp. italica）下胚轴为外植体，通过农杆菌介导的遗传转化方法将来源于杨树的Kunitz 型丝氨酸胰蛋白酶抑制剂（KTI）基因导入青花菜品系‘09LR-11’中，获得13株卡那霉素抗性植株。以KTI 特异引物对转基因植株进行PCR 检测，其中8 株为KTI 阳性植株。Southernblot 分析进一步表明，基因KTI 已成功整合到青花菜基因组中。RT-PCR 检测表明，KTI 在转基因青花菜中已成功表达。通过室内叶片离体试验和田间观察，初步证明转KTI 青花菜对小菜蛾幼虫具有一定抗性

Transformation of Broccoli with KTI Gene and the Bioassay forDiamondback Moth Resistance

KTI gene was transformed into broccoli（Brassica oleracea L. ssp. italica）genome viaAgrobacterium mediated transformation method using broccoli lines‘09LR-11’as explants. Thirteenkanamycin resistant broccoli plants were obtained. PCR analysis with primers specific to KTI geneidentified 8 KTI positive plants. Southern blotting analysis with KTI gene as probes showed that the KTIgene was integrated into broccoli genome. RT-PCR analysis indicated that the KTI gene was expressed intransgenic broccoli. Both laboratory and field tests proved that the transgenic broccoli plants were resistantto diamondback moth（Plutella xylostella larvae）.