Welcome to Acta Agronomica Sinica，

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

PDF

2117

Cited

3

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

[1]Zhang Z Y, Lin Z S, Xin Z Y. Research progress in BYDV resistance genes derived from wheat and its wild relatives. J Genet Genom, 2009, 36: 567–573
[2]Zhang Z Y, Xu J S, Xu Q J, Larkin P , Xin Z Y. Development of novel PCR markers linked to the BYDV resistance gene Bdv2 useful in wheat for marker-assisted selection. Theor Appl Genet, 2004, 109: 433–439
[3]Dangl J L, Jones J D G. Plant pathogens and integrated defence responses to infection. Nature, 2001, 411: 826–833
[4]Yi T-Y(易图永), Xie B-Y(谢丙炎), Zhang B-X(张宝玺), Gao B-D(高必达). Application of plant resistance gene analogs in cloning and mapping resistance genes .Biotechnol Bull (生物技术通报), 2002, (2): 16–20 (in Chinese with English abstract)
[5]Qin G-J(秦跟基), Li W-L(李万隆), Chen P-D(陈佩度). Update of resistance genes and resistance gene analogs in plants. J Nanjing Agric Univ (南京农业大学学报), 1999, 22(3): 102–107 (in Chinese with English abstract)
[6]Jones D A, Jones J D G. The roles of leucine-rich repeats in plants defences. Adv Bot Res, 1997, 24: 89–167
[7]Kajava A V. Structural diversity of leucine-rich repeat proteins. J Mol Biol, 1998, 227: 519–527
[8]Whitham S, McCormick S, Baker B. The N gene of tobacco confers resistance to tobacco mosaic virus in transgenic tomato. Proc Natl Acad Sci USA, 1996, 93: 8776–8781
[9]Spassova M I, Prins T W, Folkertsma R T, Klein-Lankhorst R M, Hille J, Goldbach R W, Prins M. The tomato gene Sw-5 is a member of the coiled coil, nucleotide binding, leucine-rich repeat class of plant resistance genes and confers resistance to TSWV in tobacco. Mol.Breed, 2001, 7: 151–161
[10]Ratcliff F, Harrison B D, Baulcombe D C. A similarity between viral defense and gene silencing in plants. Science, 1997, 276: 1558–1560
[11]Wang H-Z(王宏芝), Li R-F(李瑞芬), Wang G-Y(王国英), Ma R-C(马荣才), Wei J-H(魏建华). Virus induced gene silence and its application in plant gene functional genomics. Prog Nat Sci (自然科学进展), 2005, 15(1): 8–14 (in Chinese)
[12]Scofield S R, Huang L, Brandt A S, Bikram S G. Development of a virus-induced gene-silencing system for hexaploid wheat and its use in functional analysis of the Lr21-mediated leaf rust resistance pathway. Plant Physiol, 2005, 138: 2165–2173
[13]Zhang Z-Y(张增艳), Yao W-L(姚乌兰), Xin Z-Y(辛志勇). Advance in virus-induced gene silencing, a novel powerfully tools for functional analysis of plant genes. J Plant Genet Resour (植物遗传资源学报), 2006, 7(1): 100–105 (in Chinese with English abstract)
[14]Lu R, Martin-Hernandez A M, Peart J R, Malcuit I, Baulcombe D C. Virus-induced gene silencing in plants. Methods, 2003, 30: 296–303
[15]Holzberg S, Brosio P, Gross C, Pogue G P. Barley stripe mosaic virus-induced gene silencing in a monocot plant .Plant J, 2002, 30: 315–327
[16]Lu X-D(刘晓东), Zhang Z-Y(张增艳), Yao W-L(姚乌兰), Xin Z-Y(辛志勇). Implement of barley stripe mosaic virus-based induced gene silencing in wheat. Acta Agron Sin (作物学报), 2005, 31(11): 1518–1520 (in Chinese with English abstract)
[17]Zhou H B, Li S F, Deng Z Y, Wang X P, Chen T, Zhang J S, Chen S Y, Ling H Q, Zhang A M, Wang D W. Zhang X Q. Molecular analysis of three new receptor-like kinase genes from hexaploid wheat and evidence for their participation in the wheat hypersensitive response to stripe rust fungus infection. Plant J, 2007, 52: 420–434
[18]Ding X S, Schneider W L, Chaluvadi S R, Mian M A R, Nelson R S. Characterization of a Brome mosaic virus strain and its use as a vector for gene silencing in monocotyledonous hosts. Mol Plant Microbe Interactions, 2006, 19, 1229–1239
[19]Vander-Linde K, Kastner C, Kumlehn J, Kahmann R, Doehlemann G. Systemic virus-induced gene silencing allows functional characterization of maize genes during biotrophic interaction with Ustilago maydis. New Phytol, 2011, 189: 471–483
[20]Zhang Z-Y(张增艳), Ma Y-Z(马有志), Xin Z-Y(辛志勇), Chen X( 陈孝), Wu D-L(武东亮), Lin Z-S(林志珊). Analysis of the chromosome constitution of wheat lines resistant to Barley yellow dwarf virus by genomic in situ hybridization. Sci Agric Sin (中国农业科学), 1998, 31(3): 1–4 (in Chinese with English abstract)
[21]Meyers B C, Dickeman A W, Michelmore R W, Sivaramakrishnan S, Sobral B W, Young N D. Plant disease resistance genes encode members of an ancient and diverse protein family within the nucleotide-binding super family. Plant J, 1999, 20: 317–332
[1] HU Wen-Jing, LI Dong-Sheng, YI Xin, ZHANG Chun-Mei, ZHANG Yong. Molecular mapping and validation of quantitative trait loci for spike-related traits and plant height in wheat [J]. Acta Agronomica Sinica, 2022, 48(6): 1346-1356.
[2] GUO Xing-Yu, LIU Peng-Zhao, WANG Rui, WANG Xiao-Li, LI Jun. Response of winter wheat yield, nitrogen use efficiency and soil nitrogen balance to rainfall types and nitrogen application rate in dryland [J]. Acta Agronomica Sinica, 2022, 48(5): 1262-1272.
[3] LEI Xin-Hui, WAN Chen-Xi, TAO Jin-Cai, LENG Jia-Jun, WU Yi-Xin, WANG Jia-Le, WANG Peng-Ke, YANG Qing-Hua, FENG Bai-Li, GAO Jin-Feng. Effects of soaking seeds with MT and EBR on germination and seedling growth in buckwheat under salt stress [J]. Acta Agronomica Sinica, 2022, 48(5): 1210-1221.
[4] FU Mei-Yu, XIONG Hong-Chun, ZHOU Chun-Yun, GUO Hui-Jun, XIE Yong-Dun, ZHAO Lin-Shu, GU Jia-Yu, ZHAO Shi-Rong, DING Yu-Ping, XU Yan-Hao, LIU Lu-Xiang. Genetic analysis of wheat dwarf mutant je0098 and molecular mapping of dwarfing gene [J]. Acta Agronomica Sinica, 2022, 48(3): 580-589.
[5] FENG Jian-Chao, XU Bei-Ming, JIANG Xue-Li, HU Hai-Zhou, MA Ying, WANG Chen-Yang, WANG Yong-Hua, MA Dong-Yun. Distribution of phenolic compounds and antioxidant activities in layered grinding wheat flour and the regulation effect of nitrogen fertilizer application [J]. Acta Agronomica Sinica, 2022, 48(3): 704-715.
[6] LIU Yun-Jing, ZHENG Fei-Na, ZHANG Xiu, CHU Jin-Peng, YU Hai-Tao, DAI Xing-Long, HE Ming-Rong. Effects of wide range sowing on grain yield, quality, and nitrogen use of strong gluten wheat [J]. Acta Agronomica Sinica, 2022, 48(3): 716-725.
[7] YAN Yan, ZHANG Yu-Shi, LIU Chu-Rong, REN Dan-Yang, LIU Hong-Run, LIU Xue-Qing, ZHANG Ming-Cai, LI Zhao-Hu. Variety matching and resource use efficiency of the winter wheat-summer maize “double late” cropping system [J]. Acta Agronomica Sinica, 2022, 48(2): 423-436.
[8] WANG Yang-Yang, HE Li, REN De-Chao, DUAN Jian-Zhao, HU Xin, LIU Wan-Dai, GU Tian-Cai, WANG Yong-Hua, FENG Wei. Evaluations of winter wheat late frost damage under different water based on principal component-cluster analysis [J]. Acta Agronomica Sinica, 2022, 48(2): 448-462.
[9] CHEN Xin-Yi, SONG Yu-Hang, ZHANG Meng-Han, LI Xiao-Yan, LI Hua, WANG Yue-Xia, QI Xue-Li. Effects of water deficit on physiology and biochemistry of seedlings of different wheat varieties and the alleviation effect of exogenous application of 5-aminolevulinic acid [J]. Acta Agronomica Sinica, 2022, 48(2): 478-487.
[10] XU Long-Long, YIN Wen, HU Fa-Long, FAN Hong, FAN Zhi-Long, ZHAO Cai, YU Ai-Zhong, CHAI Qiang. Effect of water and nitrogen reduction on main photosynthetic physiological parameters of film-mulched maize no-tillage rotation wheat [J]. Acta Agronomica Sinica, 2022, 48(2): 437-447.
[11] MA Bo-Wen, LI Qing, CAI Jian, ZHOU Qin, HUANG Mei, DAI Ting-Bo, WANG Xiao, JIANG Dong. Physiological mechanisms of pre-anthesis waterlogging priming on waterlogging stress tolerance under post-anthesis in wheat [J]. Acta Agronomica Sinica, 2022, 48(1): 151-164.
[12] MENG Ying, XING Lei-Lei, CAO Xiao-Hong, GUO Guang-Yan, CHAI Jian-Fang, BEI Cai-Li. Cloning of Ta4CL1 and its function in promoting plant growth and lignin deposition in transgenic Arabidopsis plants [J]. Acta Agronomica Sinica, 2022, 48(1): 63-75.
[13] WEI Yi-Hao, YU Mei-Qin, ZHANG Xiao-Jiao, WANG Lu-Lu, ZHANG Zhi-Yong, MA Xin-Ming, LI Hui-Qing, WANG Xiao-Chun. Alternative splicing analysis of wheat glutamine synthase genes [J]. Acta Agronomica Sinica, 2022, 48(1): 40-47.
[14] LI Ling-Hong, ZHANG Zhe, CHEN Yong-Ming, YOU Ming-Shan, NI Zhong-Fu, XING Jie-Wen. Transcriptome profiling of glossy1 mutant with glossy glume in common wheat (Triticum aestivum L.) [J]. Acta Agronomica Sinica, 2022, 48(1): 48-62.
[15] ZHAO Hai-Han, LIAN Wang-Min, ZHAN Xiao-Deng, XU Hai-Ming, ZHANG Ying-Xin, CHENG Shi-Hua, LOU Xiang-Yang, CAO Li-Yong, HONG Yong-Bo. Genetic dissection of the bacterial blight disease resistance in super hybrid rice RILs using genome-wide association study [J]. Acta Agronomica Sinica, 2022, 48(1): 121-137.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd