大豆中一个WRKY28-like基因的克隆与功能分析  

Cloning and Functional Analysis of a WRKY28-like Gene in Soybean

作　　者：王婷婷[1] 丛亚辉 柳聚阁[1] 王宁[1] 帅琴[1] 李艳[1] 盖钧镒[1] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室/国家大豆改良中心/农业部大豆生物学与遗传育种重点实验室/江苏省现代作物生产协同创新中心,江苏南京210095

出　　处：《作物学报》2016年第4期469-481,共13页Acta Agronomica Sinica

基　　金：国家高技术研究发展计划(863计划)(2013AA102602)项目;教育部长江学者和创新团队发展计划(PCSIRT13073)项目;中央高校基本科研业务费专项资金;教育部新世纪优秀人才支持计划(NCET-12-0891);农业部大豆生物学与遗传育种创新团队和江苏省双创计划资助~~

摘　　要：WRKY转录因子参与调节植物生长发育、生物与非生物胁迫应答等多种过程,At WRKY28是拟南芥中与抗病和耐逆相关的重要转录因子。为探讨大豆中一个At WRKY28同源基因Gm WRKY28-like(Glyma.14G028900)的生物学功能,本文对该基因进行了克隆、生物信息学分析、亚细胞定位、组织表达等试验,并对其在ABA、PEG、Na Cl胁迫下的表达水平进行了分析。结果显示,GmWRKY28-like基因的编码区(CDS)为1008 bp,编码335个氨基酸。GmWRKY28-like蛋白具有保守的WRKY结构域,含有22个丝氨酸(Serine)、1个苏氨酸(Threonine)、2个酪氨酸(Tyrosine),不含跨膜结构与信号肽;进化树分析表明大豆GmWRKY28-like与菜豆(Phaseolus vulgaris)WRKY28的相似性最高;亚细胞定位显示GmWRKY28-like定位在细胞核中。该基因在根、种子中表达量很低,在真叶、花、及茎尖分生组织表达量较高。GmWRKY28-like启动子中含有多种与生物和非生物逆境胁迫应答相关的元件,如MBS、W-box、ABRE、Box-W1等,且表达受到ABA、PEG、Na Cl的诱导。此外,过表达GmWRKY28-like显著增强了拟南芥的耐盐性。WRKY transcription factors play important roles in regulating plant growth and development, and participate in plant response to abiotic and biotic stresses. Arabidopsis WRKY28 is an essential transcription factor for plant response to necrotrophic pathogen and abiotic stress. To explore the function of the homologues gene of At WRKY28 in soybean, we cloned GmWRKY28-like（Glyma.14G028900） and performed bioinformatics analysis. Its sub-cellular localization and tissue expression patterns were further investigated. We also analyzed the expression levels of Gm WRKY28-like under ABA, PEG, and Na Cl stress treatments. Our results showed that the coding DNA sequence（CDS） of GmWRKY28-like gene is 1008 bp in length, encoding 335 amino acids. GmWRKY28-like contains a conserved WRKY domain with 22 serine, one threonine and two tyrosine, without any transmembrane domain or signal peptide. Phylogenetic analysis showed that the WRKY28 from Phaseolus vulgaris was most similar to GmWRKY28-like. GmWRKY28-like was verified to be located in the nucleus. This gene expressed at low levels in root and seed while high levels in true leaf, flower and shoot apical meristem. The 1500 bp upstream region of GmWRKY28-like contains a variety of cis-elements, such as MBS, W box, ABRE, and box-W1, which are involved in the response to biotic and abiotic stresses, and its expression was induced by ABA, PEG, and NaCl treatments. In addition, overexpressing Gm WRKY28-like in Arabidopsis thaliana enhanced plant tolerance to Na Cl. This study would provide reference basis to further study the functions of GmWRKY28-like in soybean tolerance to abiotic stresses.

关 键 词：GmWRKY28 进化树分析 亚细胞定位 非生物胁迫 

分 类 号：Q943.2[生物学—植物学]