作物学报2011，Vol.37Issue(11)：2106-2110,5.DOI:10.3724/SP.J.1006.2011.02106
利用BSMV-VIGS技术快速分析小麦TNBL1基因的抗黄矮病功能
Functional Analysis of TNBL1 Gene in Wheat Defense Response to Barley yellow dwarf virus Using BSMV-VIGS Technique
摘要
Abstract
Barley yellow dwarf virus (BYDV), transmitted by at least 25 species of aphids, causes one of the most serious virus diseases of wheat worldwide. Through Cdna-AFLP analysis, we identified a Cdna fragment with 292bp expressing in the BYDV-resistant wheat-Thinopyrun intermedium translocation line YW642, but not in susceptible wheat Zhong8601. The full-length Cdna sequence of the gene, namely TNBL1, was cloned by RACE and RT-PCR methods, which encodes a putative NBS-LRR protein. This study focused on the functional analysis of TNBL1 in wheat defense to BYDV infection using Barley stripe mosaic virus (BSMV)-based virus-inducing gene silencing method. After the specific fragment of TNBL1 was added with 2 restriction-enzyme sequences by PCR, and digested and ligased with the digested BSMV-γ, the recombinant BSMW-γ:TNBLlas construct was obtained. The three components of the BSMV-VIGS vectors, BSMV-TNBLlas, BSMV-α and BSMV-β were tran-scribed in vitro, and mixed with equal quantity and inoculated onto the first and second leaves of the resistant line YW642 seed-lings at the two-leaf stage. As a result, the TNBL1 expression was obviously repressed (silenced) in YW642 treated by BSMV:TNBL1. These seedlings were further inoculated with BYDV aphids. The BYDV content was much higher in the TNVBL1-silenced YW642 plants than that in the control YW642 plants without BSMV:TNBL1 treatment. Furthermore, the TNVBL1-silenced YW642 plants were susceptible to BYDV infection with the viral symptom. These results indicated that the TNBL1 gene is an important gene positively involved in wheat defense response to BYDV infection.关键词
小麦/中间偃麦草/黄矮病/病毒诱导的基因沉默/NBS-LRRKey words
Wheat/ Barley yellow dwarf virus/ Thinopyrum intermedium/ Virus-induced gene silencing/ NBS-LRR引用本文复制引用
赵丹,赵继荣,黄茜,李宁,刘艳,黄占景,张增艳..利用BSMV-VIGS技术快速分析小麦TNBL1基因的抗黄矮病功能[J].作物学报,2011,37(11):2106-2110,5.基金项目
本研究由国家高技术发展计划(863计划)重点项目(2006AA 10A 104)资助. （863计划）
