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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (11): 2106-2110.doi: 10.3724/SP.J.1006.2011.02106

• RESEARCH NOTES • Previous Articles     Next Articles

Functional Analysis of TNBL1 Gene in Wheat Defense Response to Barley yellow dwarf virus Using BSMV-VIGS Technique

ZHAO Dan1,2,ZHAO Ji-Rong1,HUANG Xi1,2,LI Ning1,HUANG Zhan-Jing2,ZHANG Zeng-Yan1,*   

  1. 1 National Key Facility for Crop Gene Resources and Genetic Improvement, Institute of Crop Sciences / Key Laboratory of Crop Genetic and Breeding, Ministry of Agriculture, Chinese Academy of Agricultural Sciences, Beijing 100081, China; 2 College of Life Science, Hebei Normal University, Shijiazhuang 050016, China
  • Received:2011-02-18 Revised:2011-06-25 Online:2011-11-12 Published:2011-09-06
  • Contact: 张增艳, E-mail: zhangzy@mail.caas.net.cn, Tel: 010-82108781

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 addedwith 2 restriction-enzyme sequences byPCR, and digested and ligased with the digested BSMV-γ, the recombinant BSMV-γ:TNBL1as construct was obtained. The three components of the BSMV-VIGS vectors, BSMV-TNBL1as、BSMV-α and BSMV-β were transcribed in vitro, and mixed with equal quantity and inoculated onto the first and second leaves of the resistant line YW642 seedlings 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 TNBL1-silenced YW642 plants than that in the control YW642 plants without BSMV:TNBL1 treatment. Furthermore, the TNBL1-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.

Key words: Wheat, Barley yellow dwarf virus, Thinopyrum intermedium, Virus-induced gene silencing, NBS-LRR

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