Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (19): 4035-4043.doi: 10.3864/j.issn.0578-1752.2013.19.009

• PLANT PROTECTION • Previous Articles     Next Articles

Isolation and Expression Patterns of Rice WRKY80 Transcription Regulatory Protein Gene

 PENG  Xi-Xu, TANG  Xin-Ke, ZHOU  Ping-Lan, HU  Yao-Jun, DENG  Xiao-Bo, WANG  Hai-Hua   

  1. School of Life Sciences, Hunan University of Science and Technology, Xiangtan 411201, Hunan
  • Received:2013-04-10 Online:2013-10-01 Published:2013-05-31

Abstract: 【Objective】 The objective of this study is to isolate rice WRKY80, to analyze the characteristics of its coding sequence and to investigate its expression patterns in different organs and under pathogen inoculation and hormone application, thus providing a basis for its function identification. 【Method】Specific primer sequences were designed according to the annotated gene Loc_Os03g63810 released in rice genome database. RT-PCR was used to amplify WRKY80 cDNA sequence from RNA pools generated from methyl jasmonate (MeJA)-treated rice leaves. Bioinformatical tools were employed to analyze its deduced protein sequence and cis-elements in its promoter. Northern blot or real-time fluorescence quantification PCR was used to investigate its expression patterns. 【Result】 The obtained cDNA sequence of WRKY80 was 1 392 bp in length, containing an entire open reading frame of 1 164 bp, encoding a polypeptide of 387 amino acid residues consisting of one classic conserved WRKY domain with a zinc finger motif of C2H2, belonging to the WRKY subgroup Ⅱ. WRKY80 possessed an acidic C terminus with consecutive 6 glutamines and 8 threonines, an acidic region possibly responsible for transcription activating activity, and was predicted to be localized in nucleus. WRKY80 shared high identity at the amino acid level with those from monocotyledons Zea mays and Sorghum bicolor. WRKY80 was constitutively expressed in all tested organs. The transcript abundance was relatively higher in leaves, roots and panicles, next was in flowers, and less was in stems and grains. WRKY80 expression was higher in mature leaves and roots than in young leaves and roots, respectively, indicating a developmental stage-related feature. It was rapidly induced by inoculation with Magnaporthe oryzae and Rhizoctonia solani, and also by application of exogenous MeJA and ethephon, whereas salicylic acid exerted no effects on its expression. Consistent with the expression profiles was the prediction result of cis-elements in its promoter. 【Conclusion】WRKY80 possesses structure characteristics as a putative transcription factor. These findings suggest that it may be involved in defense response to the fungal pathogens and also in development regulation by jasmonic acid/ethylene-dependent signaling pathway.

Key words: WRKY transcription factor , fungal pathogen , gene isolation , gene expression pattern , Oryza sativa

[1]Rushton P J, Somssich I E. Transcriptional control of plant genes response to pathogens. Current Opinion in Plant Biology, 1998, 1(4): 311-315.

[2]Eulgem T, Rushton P J, Robatzek S, Somssich I E. The WRKY superfamily of plant transcription factors. Trends in Plant Science, 2000, 5(5): 199-206.

[3]Pandey S P, Somssich I E. The role of WRKY transcription factors in plant immunity. Plant Physiology, 2009, 150(4): 1648-1655.

[4]Ryu H S, Han M, Lee S K, Cho J I, Ryoo N, Heu S, Lee Y H, Bhoo S H, Wang G L, Hahn T R, Jeon J S. A comprehensive expression analysis of the WRKY gene superfamily in rice plants during defense response. Plant Cell Reports, 2006, 25(8): 836-847.

[5]Yu D, Chen C, Chen Z. Evidence for an important role of WRKY DNA binding proteins in the regulation of NPR1 gene expression. The Plant Cell, 2001, 13(7): 1527-1540.

[6]Deslandes L, Olivier J, Theuliéres T, Hirsch J, Feng D X, Bittner-Eddy P, Beynon J, Marco Y. Resistance to Ralstonia solanacearum in Arabidopsis thaliana is conferred by the recessive RRS1-R gene, a member of a novel family of resistance genes. Proceedings of the National Academy of Sciences of the United States of America, 2002, 99(4): 2404-2409.

[7]Eulgem T, Somssich I E. Networks of WRKY transcription factors in defense signaling. Current Opinion in Plant Biology, 2007, 10(4): 366-371.

[8]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(1): 9-26.

[9]Rice WRKY Working Group. Nomenclature report on rice WRKY’s. -Conflict regarding gene names and its solution. Rice, 2012, 5: 3.

[10]Shimono M, Sugano S, Nakayama A, Jiang C J, Ono K, Toki S, Takatsuji H. Rice WRKY45 plays a crucial role in benzothiadiazole-inducible blast resistance. The Plant Cell, 2007, 19(6): 2064-2076.

[11]Matsushita A, Inoue H, Goto S, Nakayama A, Sugano S, Hayashi N, Takatsuji H. Nuclear ubiquitin proteasome degradation affects WRKY45 function in the rice defense program. The Plant Journal, 2013, 73(2): 302-313.

[12]Qiu D, Xiao J, Ding X, Xiong M, Cai M, Cao Y, Li X, Xu C, Wang S. OsWRKY13 mediates rice disease resistance by regulating defense-related genes in salicylate- and jasmonate-dependent signaling. Molecular Plant-Microbe Interactions, 2007, 20(5): 492-499.

[13]Peng Y, Bartley L E, Chen X, Dardick C, Chern M, Ruan R, Canlas P E, Ronald P C. OsWRKY62 is a negative regulator of basal and Xa21-mediated defense against Xanthomonas oryzae pv. oryzae in rice. Molecular Plant, 2008, 1(3): 446-458.

[14]Liu X Q, Bai X Q, Qian Q, Wang X J, Chen M S, Chu C C. OsWRKY03, a rice transcriptional activator that functions in defense signaling pathway upstream of OsNPR1. Cell Research, 2005, 15: 593-603.

[15]Liu X Q, Bai X Q, Wang X J, Chu C C. OsWRKY71, a rice transcription factor, is involved in rice defense response. Journal of Plant Physiology, 2007, 164(8): 969-979.

[16]Silverman P, Seskar M, Kanter D, Schweizer P, Metraux J P, Raskin  I. Salicylic acid in rice: biosynthesis, conjugation, and possible role. Plant Physiology, 1995, 108(2): 633-639.

[17]Peng X X, Hu Y J, Tang X K, Zhou P L, Deng X B, Wang H H, Guo Z J. Constitutive expression of rice WRKY30 gene increases the endogenous jasmonic acid accumulation, PR gene expression and resistance to fungal pathogens in rice. Planta, 2012, 236(5): 1485-1498.

[18]Ricachenevsky F K, Sperotto R A, Menguer P K, Fett J P. Identification of Fe-excess-induced genes in rice shoots reveals a WRKY transcription factor responsive to Fe, drought and senescence. Molecular Biology Reports, 2010, 37(8): 3735-3745.

[19]李南羿,  柴荣耀,  郭泽建. OsWRKY80基因参与调控水稻抗病反应研究. 上海农业学报, 2009, 25(3): 14-18.

Li N Y, Chai R Y, Guo Z J. The disease resistance of rice regulated by OsWRKY80 gene. Acta Agriculturae Shanghai, 2009, 25(3): 14-18. (in Chinese)

[20]Peng X X, Tang X K, Zhou P L, Hu Y J, Deng X B, He Y, Wang H H. Isolation and expression patterns of rice WRKY82 transcription factor gene responsive to both biotic and abiotic stresses. Agricultural Sciences in China, 2011, 10(6): 893-901.

[21]Peng Y L, Shishiyama J. Temporal sequence of cytological events in rice leaves affected with Pyricularia oryzae. Canadian Journal of Botany, 1988, 66(4): 730-735.

[22]Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Analytical Biochemistry, 1987, 162(1): 156-159.

[23]Sambrook J, Fritsch E F, Maniatis T. Molecular Cloning: A Laboratory Manual. 2nd ed. New York: Cold Spring Harbor Laboratory Press, 1989.

[24]Schwechheimer C, Bevan M. The regulation of transcription factor activity in plants. Trends in Plant Science, 1998, 3(10): 378-383.

[25]Zhang J, Peng Y L, Guo Z J. Constitutive expression of pathogen- inducible OsWRKY31 enhances disease resistance and affects root growth and auxin response in transgenic rice plants. Cell Research, 2008, 18: 508-521.

[26]Wang H H, Hao J J, Chen X J, Hao Z N, Wang X, Lou Y G, Peng Y  L, Guo Z J. Overexpression of rice WRKY89 enhances ultraviolet B tolerance and disease resistance in rice plants. Plant Molecular Biology, 2007, 65(6): 799-815.

[27]Ramamoorthy R, Jiang S Y, Kumar N, Venkatesh P N, Ramachandran S. A comprehensive transcriptional profiling of the WRKY gene family in rice under various abiotic and phytohormone treatments. Plant & Cell Physiology, 2008, 49(6): 865-879.

[28]Besseau S, Li J, Palva E T. WRKY54 and WRKY70 co-operate as negative regulators of leaf senescence in Arabidopsis thaliana. Journal of Experimental Botany, 2012, 63(7): 2667-2679.

[29]Ülker B, Somssich I E. WRKY transcription factors: from DNA binding towards biological function. Current Opinion in Plant Biology, 2004, 7(5): 491-498.
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