摘要
将含有雪花莲外源凝集素基因 (GNA)的质粒pRSSGNA1通过冻融法转化到根癌土壤杆菌 (Agrobacteriumtumefaciens (SmithetTownsend)Conn)菌株LBA44 0 4中。采用叶盘法转化番茄 (LycopersiconesculentumMill.)栽培品种“C8”、“A3 9”和“A53 ” ,获得了含GNA基因的 43株转化植株。通过卡那霉素抗性鉴定、NPTⅡ基因PCR和GNA基因Southernblot分析表明 ,GNA基因已整合到番茄的基因组中。初步抗蚜虫 (MyzuspersicaeSulzer)试验证明 ,转基因番茄有一定的抗蚜虫效果。 3株转基因植株的NPTⅡ基因在自交子代中的分离比例符合 3∶1孟德尔遗传规律。
The plasmid pRSSGNA1 carried a snowdrop lectin gene (Galanthus nivalis agglutinin, GNA) under the drive of RSs_1 promoter, were successfully transferred into three tomato (Lycopersicon esculentum Mill.) cultivars, “C 8 ”, “A 39 ” and “A 53 ” via Agrobacterium_mediated transformation. Regenerated plantlets were obtained from cotyledons after preculture, shoot inducing culture and root inducing culture. Transgenic tomato plants were confirmed by the kanamycin_resistant experiment, PCR analysis and Southern blot. The preliminary results from bioassay demonstrated significant resistance of the transgenic plants to aphid (Myzus persicae Sulzer) larvae. The inheritance of selective marker gene (NPTⅡ) in 3 transgenic tomato plants is in the model of the simple Mendel's fashion in progenies of the selfing generation.
基金
武汉市科技重点攻关项目&&
关键词
番茄
蚜虫
GAN
抗虫育种
基因转化
tomato
snowdrop lectin gene (GNA)
genetic transformation
aphid
