Scientia Agricultura Sinica ›› 2016, Vol. 49 ›› Issue (12): 2245-2254.doi: 10.3864/j.issn.0578-1752.2016.12.001

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS •     Next Articles

Cloning and Characterization of Transcription Factor TaWRKY35 in Wheat (Triticum aestivum)

LIU Zi-cheng1, 2, MIAO Li-li2, WANG Jing-yi2, YANG De-long1, MAO Xin-guo1,2, JING Rui-lian2   

  1. 1College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070
    2 Institute of Crop Science, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement/Key Laboratory of Crop Germplasm and Utilization, Ministry of Agriculture, Beijing 100081
  • Received:2016-03-04 Online:2016-06-16 Published:2016-06-16

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Abstract: 【Objective】Abiotic stresses are major limitations to wheat production worldwide. Transcription factors play crucial roles in abiotic stress signaling in plants. It is predicted that there are at least 200 WRKY genes in common wheat genomes, yet only a few of them have been functionally characterized. The aim of this study is to decipher the roles of WRKY transcription factors in abiotic stress signaling and facilitate the utilization of WRKY genes in the improvement of abiotic stress tolerance in wheat.【Method】A wheat WRKY gene designated TaWRKY35 was cloned via the wheat full-length cDNA libraries. Quantitative real-time PCR was performed to identify the dynamic expression of target gene in different tissues at various developmental stages, and to characterize the transcriptional patterns responding to ABA, PEG, NaCl and low temperature treatments in wheat. To probe the subcellular location of TaWRKY35, the construct encoding TaWRYK35::GFP fusion protein was transferred into wheat protoplast by PEG mediated method. To characterize the function of TaWRYK35, target gene driven by the 35S promoter was delivered into Arabidopsis by Agrobacterium mediated method. 【Result】The cDNA of TaWRYK35 contains an 1 134 bp open reading frame, encoding a 377- amino acid protein. TaWRYK35 possesses a typical WRKY domain in the N-terminal and a C2HC type zinc finger domain in the C-terminal, belonging to group III of WRKY family. In 32 hexaploid wheat materials of highly polymorphic, TaWRKY35 coding region sequence is very conservative. The dynamic expression of TaWRKY35 was identified in different tissues at various developmental stages, and the highest expression occurred in the root base of seedling, while the lowest expression was observed in seedling leaf. Furthermore, its transcript was inducible by ABA, PEG, NaCl and low temperature treatments, yet the expression patterns to difference stress varied significantly. Subcellular localization indicated that TaWRKY35 specifically located in the nucleus. Overexpression of TaWRKY35 resulted in enhanced tolerance to high salinity, supported by improved cell membrane stability and survival rate relative to wild type Arabidopsis. 【Conclusion】TaWRKY35 transcription factor contains a WRKY and a C2HC type zinc finger domain, belonging to subgroup III of the WRKY family. The expression of TaWRKY35 occurs in different tissues at various developmental stages, and TaWRKY35 is an abiotic stress responsive gene. Overexpression of TaWRKY35 confers remarkably enhanced tolerance to high salinity.

Key words: common wheat, WRKY transcription factor gene, abiotic stress, salt tolerance

[1]    Suzuki N, Rivero R M, Shulaev V, Blumwald E, Mittler R. Abiotic and biotic stress combinations. New Phytologist, 2014, 203: 32-43.
[2]    Mickelbart M V, Hasegawa P M, Bailey-Serres J. Genetic mechanisms of abiotic stress tolerance that translate to crop yield stability. Nature Reviews Genetics, 2015, 16: 237-251.
[3]    Ishiguro S, Nakamura K. Characterization of a cDNA encoding a novel DNA-binding protein, SPF1, that recognizes SP8 sequences in the 5' upstream regions of genes coding for sporamin and beta-amylase from sweet potato. Molecular and General Genetics, 1994, 244: 563-571.
[4]    Eulgem T, Rushton P J, Robatzek S, Somssich I E. The WRKY superfamily of plant transcription factors. Trends in Plant Science, 2000, 5: 199-206.
[5]    Maeo K, Hayashi S, Kojima-Suzuki H, Morikami A, Nakamura K. Role of conserved residues of the WRKY domain in the DNA-binding of tobacco WRKY family proteins. Bioscience, Biotechnology, and Biochemistry, 2001, 65: 2428-2436.
[6]    Wu K L, Guo Z J, Wang H H, Li J. The WRKY family of transcription factors in rice and Arabidopsis and their origins. DNA Research, 2005, 12: 9-26.
[7]    Ciolkowski I, Wanke D, Birkenbihl R P, Somssich I E. Studies on DNA-binding selectivity of WRKY transcription factors lend structural clues into WRKY-domain function. Plant Molecular Biology, 2008, 68: 81-92.
[8]    Rushton P J, Somssich I E, Ringler P, Shen Q J. WRKY transcription factors. Trends in Plant Science, 2010, 15: 247-258.
[9]    Rushton D L, Tripathi P, Rabara R C, Lin J, Ringler P, Boken A K, Langum T J, Smidt L, Boomsma D D, Emme N J, Chen X, Finer J J, Shen Q J, Rushton P J. WRKY transcription factors: key components in abscisic acid signalling. Plant Biotechnology Journal, 2012, 10: 2-11.
[10]   Li C, Lü J, Zhao X, Ai X, Zhu X, Wang M, Zhao S, Xia G. TaCHP: a wheat zinc finger protein gene down-regulated by abscisic acid and salinity stress plays a positive role in stress tolerance. Plant Physiology, 2010, 154: 211-221.
[11]   Li H, Xu Y, Xiao Y, Zhu Z, Xie X, Zhao H, Wang Y. Expression and functional analysis of two genes encoding transcription factors, VpWRKY1 and VpWRKY2, isolated from Chinese wild Vitis pseudoreticulata. Planta, 2010, 232: 1325-1337.
[12]   Jiang Y, Deyholos M K. Functional characterization of Arabidopsis NaCl-inducible WRKY25 and WRKY33 transcription factors in abiotic stresses. Plant Molecular Biology, 2009, 69: 91-105.
[13]   Li S, Fu Q, Chen L, Huang W, Yu D. Arabidopsis thaliana WRKY25, WRKY26, and WRKY33 coordinate induction of plant thermotolerance. Planta, 2011, 233: 1237-1252.
[14]   Li S, Fu Q, Huang W, Yu D. Functional analysis of an Arabidopsis transcription factor WRKY25 in heat stress. Plant Cell Reports, 2009, 28: 683-693.
[15]   Sun J, Hu W, Zhou R, Wang L, Wang X, Wang Q, Feng Z, Li Y, Qiu D, He G, Yang G. The Brachypodium distachyon BdWRKY36 gene confers tolerance to drought stress in transgenic tobacco plants. Plant Cell Reports, 2015, 34: 23-35.
[16]   Zhou Q Y, Tian A G, Zou H F, Xie Z M, Lei G, Huang J, Wang C M, Wang H W, Zhang J S, Chen S Y. Soybean WRKY-type transcription factor genes, GmWRKY13, GmWRKY21, and GmWRKY54, confer differential tolerance to abiotic stresses in transgenic Arabidopsis plants. Plant Biotechnology Journal, 2008, 6: 486-503.
[17]   Wu X, Shiroto Y, Kishitani S, Ito Y, Toriyama K. Enhanced heat and drought tolerance in transgenic rice seedlings overexpressing OsWRKY11 under the control of HSP101 promoter. Plant Cell Reports, 2009, 28: 21-30.
[18]   Raineri J, Wang S, Peleg Z, Blumwald E, Chan R L. The rice transcription factor OsWRKY47 is a positive regulator of the response to water deficit stress. Plant Molecular Biology, 2015, 88: 401-413.
[19]   Okay S, Derelli E, Unver T. Transcriptome-wide identification of bread wheat WRKY transcription factors in response to drought stress. Molecular Genetics and Genomics, 2014, 289: 765-781.
[20]   Wang X, Zeng J, Li Y, Rong X, Sun J, Sun T, Li M, Wang L, Feng Y, Chai R, Chen M, Chang J, Li K, Yang G, He G. Expression of TaWRKY44, a wheat WRKY gene, in transgenic tobacco confers multiple abiotic stress tolerances. Frontiers in Plant Science, 2015, 6: 615.
[21]   Wang C, Deng P, Chen L, Wang X, Ma H, Hu W, Yao N, Feng Y, Chai R, Yang G, He G. A wheat WRKY transcription factor TaWRKY10 confers tolerance to multiple abiotic stresses in transgenic tobacco. PLoS One, 2013, 8: e65120.
[22]   Niu C F, Wei W, Zhou Q Y, Tian A G, Hao Y J, Zhang W K, Ma B, Lin Q, Zhang Z B, Zhang J S, Chen S Y. Wheat WRKY genes TaWRKY2 and TaWRKY19 regulate abiotic stress tolerance in transgenic Arabidopsis plants. Plant Cell and Environment, 2012, 35: 1156-1170.
[23]   Qin Y, Tian Y, Liu X. A wheat salinity-induced WRKY transcription factor TaWRKY93 confers multiple abiotic stress tolerance in Arabidopsis thaliana. Biochemical and Biophysical Research Communications, 2015, 464: 428-433.
[24]   Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 2001, 25: 402-408.
[25]   Pandey S P, Somssich I E. The role of WRKY transcription factors in plant immunity. Plant Physiology, 2009, 150: 1648-1655.
[26]   Zou C, Jiang W, Yu D. Male gametophyte-specific WRKY34 transcription factor mediates cold sensitivity of mature pollen in Arabidopsis. Journal of Experimental Botany, 2010, 61: 3901-3914.
[27]   Wang F, Hou X, Tang J, Wang Z, Wang S, Jiang F, Li Y. A novel cold-inducible gene from Pak-choi (Brassica campestris ssp. chinensis), BcWRKY46, enhances the cold, salt and dehydration stress tolerance in transgenic tobacco. Molecular Biology Reports, 2012, 39: 4553-4564.
[28]   Xiong X, James V A, Zhang H, Altpeter F. Constitutive expression of the barley HvWRKY38 transcription factor enhances drought tolerance in turf and forage grass (Paspalum notatum Flugge). Molecular Breeding, 2009, 25: 419-432.
[29]   Xu Y, Hu W, Liu J, Zhang J, Jia C, Miao H, Xu B, Jin Z. A banana aquaporin gene, MaPIP1;1, is involved in tolerance to drought and salt stresses. BMC Plant Biology, 2014, 14: 59.
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