CAJ | 학술논문

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。
구건한채적병독유도기인침묵（ Virus induced gene silencing ，VIGS）체계，지재건립한채적쾌속기인공능험증적방법。통과RT-PCR확증출면화、대두、의남개적1-탈양목동당-5-린산합성매（Cloroplasto alterados，CLA）기인편단，병장기분별구건도연초취렬병독（ Tobacco rattle virus ，TRV）개도적VIGS재체，통과농간균GV3101침염한채병관찰한채적표형변화。결과현시，3충불동적CLA기인소침염적한채균출현불동정도적백화현상，반정량RT-PCR검측현시CLA적mRNA피현저강해，표명성공구건료한채적VIGS체계。
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 .