结缕草转录因子基因ZjDREB4.1克隆和逆境表达模式

doi:10.13560/j.cnki.biotech.bull.1985.2017.02.012

摘要/Abstract

摘要： 以日本结缕草（Zoysia japonica）‘Meyer’品种为材料,克隆获得了1个DREB（dehydration responsive element binding）类转录因子基因,命名为ZjDREB4.1。该基因编码区长651 bp,编码216个氨基酸,推测的蛋白质ZjDREB4.1分子量为22.9 kD,等电点pI为5.74,第50-110位氨基酸组成一个典型的AP2 保守结构域。在基因的系统发生树中,ZjDREB4.1蛋白与拟南芥AtTINY蛋白和玉米ZmDBF2蛋白聚为一支,属于DREB亚家族A-4组。在叶组织中ZjDREB4.1为组成型表达,受低温诱导上调表达,在干旱和高盐胁迫下表达先下调后恢复至正常水平。

关键词: 转录因子, DREB, 半定量RT-PCR, 非生物胁迫

Abstract: A novel DREB（dehydration responsive element binding）transcription factor gene,designated as ZjDREB4.1,was isolated from cultivar ‘Meyer’ of Zoysia japonica. The open reading frame（ORF）of ZjDREB4.1 was 651 bp in length,encoding 216 amino acid residues. The putative protein ZjDREB4.1 was 22.9 kD in molecular weight and with a theoretical isoelectric point（pI）of 5.74. Amino acids from the 50^th to the 110^th were predicted to build up a conserved AP2 domain. The phylogenetic tree analysis showed that ZjDREB4.1 was grouped with AtTINY of Arabidopsis thaliana and ZmDBF2 of maize,belonging to the A-4 group of DREB subfamily. ZjDREB4.1 was expressed constitutively in leaves. The expression was up-regulated under cold stress,and was first down-regulated then returned to normal level under drought and salt stresses.

Key words: transcription factor, DREB, semi-quantitative PCR, abiotic stress

李京, 吴奇, 张琳婕, 李旭婷, 周敏琪, 韦善君. 结缕草转录因子基因ZjDREB4.1克隆和逆境表达模式[J]. 生物技术通报, 2017, 33(2): 80-88.

LI Jing, WU Qi, ZHANG Lin-jie, LI Xu-ting, ZHOU Min-qi, WEI Shan-jun. Cloning and Expression Profiles of a Transcription Factor Gene ZjDREB4.1 in Zoysia japonica Under Adversity[J]. Biotechnology Bulletin, 2017, 33(2): 80-88.

参考文献

[1] 任继周. 草坪业是我国全民共有、全民共建、全民共享的伟大事业——在中国草学会草坪专业委员会第七届全国代表大会暨十一届学术研讨会上的发言[J]. 草地学报, 2008, 16（6）：545-546.
[2] 单华佳, 李梦璐, 孙彦, 等. 近10年中国草坪业发展现状[J]. 草地学报, 2003, 21（2）：222-227.
[3] 董厚德, 宫莉君, 等. 中国结缕草生态学及其资源开发与应用[M]. 北京：中国林业出版社, 2001, 105-113.
[4] 王艳, 李建龙, 邓蕾, 等. 4 种结缕草在南京地区的田间表现及耐寒性差异[J]. 中国草地学报, 2009, 31（6）：34-38.
[5] Barah P, Jayavelu ND, Rasmussen S, et al. Genome-scale cold stress response regulatory networks in ten Arabidopsis thaliana ecotypes[J]. BMC Genomics, 2013, 14：722.
[6] Huang GT, Ma SL, Bai LP, et al. Signal transduction during cold, salt, and drought stresses in plants[J]. Molecular Biology Reports, 2012, 39（2）：969-987.
[7] Nakashima K, Yamaguchi-Shinozaki K, Shinozaki K. The transcriptional regulatory network in the drought response and its crosstalk in abiotic stress responses including drought, cold, and heat[J]. Front Plant Sci, 2014, 5：170.
[8] Dietz KJ, Vogel MO, Viehhauser A. AP2/EREBP transcription factors are part of gene regulatory networks and integrate metabolic, hormonal and environmental signals in stress acclimation and retrograde signalling[J]. Protoplasma, 2010, 245（1）：3-14.
[9] Hu W, Yang H, Yan Y, et al. Genome-wide characterization and analysis of bZIP transcription factor gene family related to abiotic stress in cassava[J]. Sci Rep, 2016, 6：22783.
[10] Roy S. Function of MYB domain transcription factors in abiotic stress and epigenetic control of stress response in plant genome[J]. Plant Signal Behav, 2016, 11（1）：e1117723.
[11] Chen L, Song Y, Li S, et al. The role of WRKY transcription factors in plant abiotic stresses[J]. Biochim Biophys Acta, 2012, 1819（2）：120-128.
[12] Nakashima K, Takasaki H, Mizoi J, et al. NAC transcription factors in plant abiotic stress responses[J]. Biochim Biophys Acta, 2011, 1819（2）：97-103.
[13] Sakuma Y, Liu Q, Dubouzet JG, et al. DNA-binding specificity of the ERF/AP2 domain of Arabidopsis DREBs, transcription factors involved in dehydration- and cold- inducible gene expression[J]. Biochemical and Biophysical Research Communications, 2002, 290（3）：998-1009.
[14] Chen J, Xia X, Yin W. Expression profiling and functionalcharacterization of a DREB2-type gene from Populus euphratica[J]. Biochemical and Biophysical Research Communications, 2009, 378（3）：483-487.
[15] Zhao T, Liang D, Wang P, et al. Genome-wide analysis and expression profiling of the DREB transcription factor gene family in Malus under abiotic stress[J]. Mol Genet Genomics, 2012, 287（5）：423-436.
[16] Egawa C, Kobayashi F, Ishibashi M, et al. Differential regulation of transcript accumulation andalternative splicing of a DREB2 homolog under abiotic stress conditions in common wheat[J]. Genes and Genetic Systems, 2006, 81（2）：77-91.
[17] Yang Y, Wu J, Zhu K, et al. Identification and characterization of two chrysanthemum（Dendronthema × moriforlium）DREB genes, belonging to the AP2/EREBP family[J]. Mol Biol Rep, 2009, 36（1）：71-81.
[18] Zhao H, Bughrara SS. Isolation and characterization of cold-regulated transcriptional activator LpCBF3 gene from perennial ryegrass（Lolium perenne L.）[J]. Mol Genet Genomics, 2008, 279（6）：585-594.
[19] Sun S, Yu JP, Chen F, et al. TINY, a dehydration-responsive element（DRE）-binding protein-like transcription factor connecting the DRE- and ethylene-responsive element-mediated signaling pathways in Arabidopsis[J]. J Biol Chem, 2008, 283（10）：6261-6271.
[20] Wei G, Pan Y, Lei J, et al. Molecular cloning, phylogenetic analysis, expressional profiling and in vitro studies of TINY2 from Arabidopsis thaliana[J]. J Biochem Mol Biol, 2005, 38（4）：440-446.
[21] Huang B, Liu JY. Cloning and functional analysis of the novel gene GhDBP3 encoding a DRE-binding transcription factor from Gossypium hirsutum[J]. Biochim Biophys Acta, 2006, 1759（6）：263-269.
[22] Kizis D, Pagès M. Maize DRE-binding proteins DBF1 and DBF2 are involved in rab17 regulation through the drought-responsive element in an ABA-dependent pathway[J]. Plant J, 2002, 30（6）：679-689.
[23] 黄方, 何慧, 迟英俊, 等. 大豆GmTINY1基因的克隆与表达分析[J]. 作物学报, 2009, 35（12）：2174-2179.
[24] Wilson K, Long D, Swinburne J, et al. A dissociation insertion causes a semidominant mutation that increases expression of TINY, an Arabidopsis gene related to APETALA2[J]. Plant Cell, 1996, 8（4）：659-671.
[25] Liu XQ, Liu CY, Guo Q, et al. Mulberry transcription factor MnDREB4A confers tolerance to multiple abiotic stresses in transgenic tobacco[J]. PLoS One, 2015, 10（12）：e0145619.
[26] Wei S, Du Z, Gao F, et al. Global transcriptome profiles of ‘Meyer’ zoysiagrass in response to cold stress[J]. PLoS One, 2015, 10（6）：e0131153.
[27] 冯勋伟, 才宏伟. 结缕草CBF 基因的同源克隆及其转基因拟南芥的抗寒性验证[J]. 作物学报, 2014, 40（9）：1572-1578.
[28] Wang Z, Zhang F, Xuan JP, et al. Isolation and expression profiles of the ZjDREB1 gene encoding a DRE-binding transcription factor from zoysiagrass Zoysia japonica[J]. Journal of Horticultural Science and Biotechnology, 2012, 87（1）：77-83.
[29] 可祥, 农钧琇, 石大林, 等. 日本结缕草’胶东青’DREB2. 2基因克隆及表达模式研究[J]. 生物技术通报, 2016, 32（1）：115-123.
[30] Murray MG, Thompson WF. Rapid isolation of high molecular weight plant DNA[J]. Nucleic Acids Res, 1980, 8（19）：4321-4325.
[31] Lata C, Prasad M. Role of DREBs in regulation of abiotic stress responses in plants[J]. J Exp Bot, 2011, 62（14）：4731-4748.
[32] Agarwal PK, Agarwal P, Reddy MK, et al. Role of DREB transcription factors in abiotic and biotic stress tolerance in plants[J]. Plant Cell Rep, 2006, 25（12）：1263-1274.
[33] Chen Y, Yang J, Wang Z, et al. Gene structures, classification, and expression models of the DREB transcription factor subfamily in Populus trichocarpa[J]. The Scientific World Journal, 2013, 2013：954640.
[34] Tian XH, Li XP, Zhou HL, et al. OsDREB4 genes in rice encode AP2-containing proteins that bind specifically to the dehydration-responsive element[J]. Acta Botanica Sinica, 2005, 47（4）：467-476.
[35] Medina J, Bgarues M, Terol J, et al. The Arabidopsis CBF gene family is composed of three genes encoding AP2 domain containing proteins whose expression is regulated by low temperature but not by ABA or dehydration[J]. Plant Physiology, 1999, 119（2）：463-470.
[36] 谢登雷, 崔江慧, 常金华. 高粱中SbDREB2基因的克隆与表达分析[J]. 作物学报, 2013, 39（8）：1352-1359.
[37] Allen MD, Yamasaki K, Ohme-Takagi M, et al. A novel mode of DNA recognition by a β-sheet revealed by the solution structure of the GCC-box binding domain in complex with DNA[J]. EMBO J, 1998, 17（18）：5484-5496.
[38] Hao D, Ohme-Takagi M, Sarai A. Unique mode of GCC box recognition by the DNA-binding domain of ethylene responsive element-binding factor（ERF domain）in plants[J]. J Biol Chem, 1998, 273（41）：26857-26861.