蔊菜病毒诱导基因沉默体系构建  

Virus-induced Gene Silencing in Rorippa indica Hiern

作　　者：张怡[1] 徐克东[1] 杨松[1] 李豪[1] 曹鹏[1] 魏森[1] 韦丹丹[1] 李成伟[1] 

机构地区：[1]周口师范学院植物遗传与分子育种重点实验室,河南周口466001

出　　处：《华北农学报》2013年第6期65-70,共6页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(31071807;31272168)

摘　　要：构建蔊菜的病毒诱导基因沉默(Virus induced gene silencing,VIGS)体系,旨在建立蔊菜的快速基因功能验证的方法。通过RT-PCR扩增出棉花、大豆、拟南芥的1-脱氧木酮糖-5-磷酸合成酶(Cloroplasto alterados,CLA)基因片段,并将其分别构建到烟草脆裂病毒(Tobacco rattle virus,TRV)介导的VIGS载体,通过农杆菌GV3101侵染蔊菜并观察蔊菜的表型变化。结果显示,3种不同的CLA基因所侵染的蔊菜均出现不同程度的白化现象,半定量RT-PCR检测显示CLA的mRNA被显著降解,表明成功构建了蔊菜的VIGS体系。Virus-induced gene silencing system of Rorippa indica Hiern was constructed ,which will be used in functional analysis of R.indica genes.Cloroplasto alterados( CLA) gene fragments of cotton ,soybean and Arabidop-sis were obtained by using RT-PCR.The cloned CLA fragments were constructed into tobacco rattle virus vector (pYY13).The constructed vectors were transformed into Agrobacterium strain GV3101,which was used to inoculate R.indica plants.All the infected plants showed photobleaching symptom with different degrees .RT-PCR of target genes further confirmed the knock-down of CLA.The established VIGS system will contribute the further study of R. indica genes in the future research .

关 键 词：病毒诱导的基因沉默 蔊菜 CLA基因 烟草脆裂病毒 

分 类 号：Q78[生物学—分子生物学]