大白菜的马铃薯蛋白酶抑制剂基因转化及抗菜青虫性的鉴定

Genetic Transformation of Chinese Cabbage with a Inducible Potato pin n Gene and the Bioassay for Pieris rapae L. Resistance

以大白菜(B.campestrtis ssp.pekinensis)‘北京80号’生长3 d无菌苗的带柄子叶为外植体,通过根癌农杆菌(Agrobacterium tumefaciens)介导的叶盘法导入马铃薯蛋白酶抑制剂基因(pin Ⅱ)。通过抗性株的真叶在不定芽诱导培养基上的再生, 证实了嵌合体的存在,并通过此方法来消除嵌合体。获得的转基因植株经PCR、PCR-Southern杂交以及植物基因组Southern杂交证实,目的基因已经整合入植物基因组中。对菜青虫(Pieris rapae L.) 进行了转基因大白菜叶片的连续离体饲喂,结果表明:受试的转基因大白菜对菜青虫的生长发育有较明显的抑制作用。

Cotyledons with petiole from aseptic seedlings of Chinese cabbage ( B. campestris ssp. pekinensis, cultivar ' Beijing 80 ' ) were used as explants for in vitro transformation using Agrobacterium tumefaciens C58 earring plamid pBBBasta-pinII-bar. Via series concentration selection of PPT, the trans-genie resistant plants were got, and showed high Basta resistance when transplanted to the greenhouse. It was proved that by shoot regeneration from young leaf disc of the survived regenerated plant, the chimera status of the plants can be eliminated. Transgenic plants were confirmed by PCR, PCR-Southern, and genomic Southern blotting which showed that the bar and pin II genes were co-integrated into the plant genome. Continuous insect feeding trials using larvae of Pieris rapae L. revealed that larvae fed with transgenic leaf tissue had a higher mortality, and the developments of them were retarded compared with those larvae fed with non-transgenic control plants.