大豆转录因子GmMYB52的克隆、表达及结合功能分析  

Isolation, Expression and Binding Function Analysis of the Transcription Factor GmMYB52 in Soybean

作　　者：许玲[1] 王元琮[1] 何晓兰[1] 黄益洪[1] 徐照龙[1] 邵宏波[1] 张大勇[1] XU Ling;WANG Yuan-Cong;HE Xiao-Lan;HUANG Yi-Hong;XU Zhao-Long;SHAO Hong-Bo;ZHANG Da-Yong(Institute of Agricultural Resources and Environment, Jiangsu Academy of Agricultural Sciences / Salt-soil Agricultural Research Center, Nanjing 210014, China)

机构地区：[1]江苏省农业科学院农业资源与环境研究所/盐土农业研究中心,江苏南京210014

出　　处：《作物学报》2017年第10期1458-1467,共10页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31600211;31101166);江苏省自然科学基金项目(BK20151364);江苏省农业科技自主创新项目(CX(15)1005)资助~~

摘　　要：MYB类转录因子在植物的生长发育和逆境响应中有重要的调控作用。依据课题组前期获得的盐胁迫相关的数字表达谱(DGEP)数据,获得盐胁迫响应显著上调基因GmMYB52,利用RT-PCR方法从栽培大豆(Williams 82)中克隆该基因片段,并与已公布的Williams 82基因组数据库序列比对,该基因与GmMYB52序列一致。生物信息学分析表明,GmMYB52编码区(CDS)全长1083 bp,编码360个氨基酸。其编码的氨基酸序列具有MYB类转录因子的共同特征,其距N端110~160氨基酸残基处有MYB结构域;系统进化树分析表明,该基因编码的蛋白与GmMYB62、拟南芥AtMYBSt1、苜蓿Mt MYB52、水稻Os MYBS3及木豆Cc MYB-like protein J的亲缘关系最近;实时荧光定量PCR结果表明,大豆根部GmMYB52的转录水平受外界非生物逆境的调控,用脱落酸(ABA)和低温(4℃)处理后12 h明显上调,用氯化钠和PEG处理0.5~24.0 h后检测到GmMYB52的转录水平在50%~600%区间内呈现出先上调后下调再上调的趋势。GmMYB52为组成型表达,在大豆的幼苗期和开花期表达较多,在成熟期表达相对较低。GmMYB52在茎叶与开花期的花中表达较强,在根的表达较弱,而在成熟期的豆荚中几乎不表达。亚细胞定位的结果表明,GmMYB52定位于细胞核,符合典型转录因子的定位特征。酵母杂交系统检测表明,GmMYB52具有转录激活特征,并且能够与MYB相关顺式作用元件基序相结合。本研究结果表明,GmMYB52编码典型的MYB转录因子,具有转录激活活性及DNA结合活性,在大豆中的表达可能与大豆的非生物胁迫和ABA信号转导途径有关,推测其可能参与了大豆对非生物胁迫的响应。The MYB type transcription factors are involved in plant development and response to abiotic stress.GmMYB52was significantly up-regulated after salt treatment.In order to gain more information about GmMYB52,GmMYB52of Williams82was cloned by RT-PCR.Bioinformatic analysis showed the CDS of GmMYB52was1083bp,encoding360amino acid residues.A MYB domain was found in the region of110to160amino acid residues from N-terminal.Blast results showed that GmMYB52are highly homologous to GmMYB62,AtMYBSt1from Arabidopsis,MtMYB52from Medicago sativa,OsMYBS3from Oryza sativa,and CcMYB-like protein J from Cajanus cajan.Quantitative PCR(qPCR)results indicated that the transcription level of GmMYB52was upregulated under ABA and low temperature stresses,under salt,drought and cold stresses,show a bimodal pattern.GmMYB52was nearly expressed in all detected tissues,except in pods at maturing stage,and its expression level was relatively higher at seedling or flowering stages than those at maturing stage.The transcription level of GmMYB52was high in stem,leaf,and flower,during seedling and blooming stages,and low in root and pods during maturity stage.Subcellular localization results showed that GmMYB52was located in the nucleus which is in agreement with the localization characteristics of typical transcription factors.Yeast hybrid assay indicated that GmMYB52had transcriptional activation functions and could bind to several MYB cis-acting element motifs.In conclusion GmMYB52is a typical1R-MYB transcription factor,and able to bind MYB cis-acting element motifs.We speculate GmMYB52is involved in response to the abiotic stress and ABA signal transduction pathway.

关 键 词：大豆 GmMYB52 表达分析 转录激活功能 结合功能 

分 类 号：Q943.2[生物学—植物学] S565.1[农业科学—作物学]