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利用西瓜花叶病毒(Watermelonmosaicvirus,WMV)外壳蛋白(CP)基因、西葫芦黄花叶病毒(Zucchiniyellowmosaicvirus,ZYMV)复制酶(NIb)基因和黄瓜花叶病毒(Cucumbermosaicvirus,CMV)复制酶基因构建三价植物表达载体,根癌农杆菌(Agrobacteriumtumefaciens)介导法转化西瓜(Citrulluslanatus)子叶外植体获得再生植株。经PCR和Southern-blot检测,证明目的基因成功地导入西瓜植株,并能够在后代植株中遗传,转化效率约为1.7‰。在温室和大田对T2、T3代转基因株系进行了病毒接种实验和抗病性筛选鉴定,转基因西瓜株系表现出感病、抗病、免疫和症状恢复等不同的类型。其中BH1-7株系的T3代植株对ZYMV和WMV的抗病能力普遍达到中等水平以上,并保持了受体西瓜品系原有的优良农艺性状。
Three-valent plant expression was constructed using the coat protein (CP) gene of Watermelon mosaic virus (WMV), the Zb chickens mosaic virus (ZYMV) replicase (NIb) gene and the cucumber mosaic virus (CMV) Vector, Agrobacterium tumefaciens mediated transformation of Citrullus lanatus cotyledon explants to obtain regenerated plants. PCR and Southern-blot showed that the target gene was successfully introduced into watermelon plants and was inherited in progeny plants. The transformation efficiency was about 1.7 ‰. In the greenhouse and the field of T2, T3 generation of transgenic lines were vaccinated experiments and screening of disease resistance, transgenic watermelon lines showed susceptible, disease resistance, immune and recovery of different types of symptoms. Among them, the T3 plants of BH1-7 plants generally reached above the medium level of resistance to ZYMV and WMV and maintained the original agronomic traits of recipient watermelon lines.