蔊菜病毒诱导基因沉默体系构建

doi:10.7668/hbnxb.2013.06.011

华北农学报 ›› 2013, Vol. 28 ›› Issue (6): 65-70. doi: 10.7668/hbnxb.2013.06.011

所属专题： 生物技术；

蔊菜病毒诱导基因沉默体系构建

张怡, 徐克东, 杨松, 李豪, 曹鹏, 魏森, 韦丹丹, 李成伟

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

收稿日期:2013-07-24 出版日期:2013-12-28
通讯作者: 李成伟(1972－),男,河南民权人,教授,博士,主要从事植物与病原体互作和抗性分子育种研究。
作者简介:张怡(1985－),女,河南汝阳人,实验师,在读硕士,主要从事植物与微生物互作研究。
基金资助:
国家自然科学基金项目(31071807;31272168)

ViRus-induced Gene Silencing in RoRippa indica HieRn

ZHANG Yi, XU Ke-dong, YANG Song, LI Hao, CAO Peng, WEI Sen, WEI Dan-dan, LI Cheng-wei

Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou 466001, China

Received:2013-07-24 Published:2013-12-28

摘要/Abstract

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。

关键词: 病毒诱导的基因沉默, 蔊菜, CLA基因, 烟草脆裂病毒

Abstract: 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.

Key words: ViRus induced gene silencing, RoRippa indica HieRn, CLA gene, Tobacco Rattle viRus

中图分类号:

S636.9

张怡, 徐克东, 杨松, 李豪, 曹鹏, 魏森, 韦丹丹, 李成伟. 蔊菜病毒诱导基因沉默体系构建[J]. 华北农学报, 2013, 28(6): 65-70. doi: 10.7668/hbnxb.2013.06.011.

ZHANG Yi, XU Ke-dong, YANG Song, LI Hao, CAO Peng, WEI Sen, WEI Dan-dan, LI Cheng-wei. ViRus-induced Gene Silencing in RoRippa indica HieRn[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2013, 28(6): 65-70. doi: 10.7668/hbnxb.2013.06.011.

http://www.hbnxb.net/CN/Y2013/V28/I6/65

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