转WMV-2CP基因黄瓜植株的再生

Regeneration of Transgenic Cucumber Plants with a WMV-2 CP Gene

以黄瓜无菌苗子叶切段为外植体 ,通过叶盘转化法与根瘤农杆菌进行共培养建立了黄瓜的转基因系统。农杆菌菌株为LBA44 0 4,内含双元载体pBPMWMV。该质粒载体带有一个npt Ⅱ基因 (筛选具有卡那霉素抗性的植株 )和一个WMV 2CP基因。抗卡那霉素 (Kanr)的黄瓜植株经DNA分子点杂交、PCR检测以及Southernblot证实 ,外源的WMV 2CP基因确实已导入黄瓜细胞且能稳定地遗传到子一代。对WMV 2CP基因在子一代的分离进行了统计。获得的转基因子一代植株对WMV 2表现出较强的抗性 ,可以延迟发病时间 。

Cotyledon segments of Cucumis sativus L. were used as explants for transformation mediated by Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pBPMWMV containing an npt Ⅱgene and a WMV 2 CP gene (Fig.1).Transgenic plants were obtained and segregation of WMV 2 CP gene in R1 transgenic plants was analysed (Table 1). Successful genetic transformation was confirmed by DNA dot hybridization, PCR assay and Southern blot analysis (Plate I). The incidence of virus disease was low and late on transgenic plants (Plate I,Fig.4 and Table 2),thus showing that it had a certain resistance to relevant viruses.