浙江农业学报

• 作物科学 •    下一篇

水稻WRKY转录因子OsWRKY71的cDNA分离和表达分析

  

  1. 1中国计量学院 生命科学学院, 浙江 杭州 310018;2浙江省农业科学院 浙江省植物有害生物防控重点实验室,浙江 杭州310021;3浙江大学 生物技术研究所,浙江 杭州 310058
  • 出版日期:2014-11-25 发布日期:2014-12-02

Isolation and expression analysis of OsWRKY71 transcription factor in rice

  1. 1 College of Life Science, China Jiliang University, Hangzhou 310018, China;2 State Key Laboratory Breeding Base for Zhejiang Sustainable Pest and Disease Control, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China;3 Institute of Biotechnology, Zhejiang University, Hangzhou 310058, China
  • Online:2014-11-25 Published:2014-12-02

摘要: WRKY蛋白是植物中最大的转录因子家族之一,对植物的生长发育具有重要调控作用。利用SMART\|RACE\|PCR技术分离获得水稻WRKY转录因子基因(OsWRKY71)的全长cDNA序列,并进行了相关的生物信息学分析。分离到的水稻WRKY转录因子cDNA全长为1 245 bp(GenBank登录号KJ137000),开放阅读框1 047 bp,编码348个氨基酸,分子量为3828 kDa,OsWRKY71具有WRKY转录因子家族典型的保守结构域,属于第Ⅱ组WRKY蛋白家族。系统进化分析表明,OsWRKY71氨基酸序列与禾本科作物小麦的亲缘关系最近,其中与小麦序列相似性为69%,和大麦的序列相似性为68%。荧光定量PCR检测表明,OsWRKY71在孕穗期剑叶中表达丰度最高,根中最低,具有表达的空间差异性。抗病品种‘IR28’在受到稻曲菌诱导的初期,OsWRKY71基因表达呈现先升高后下降趋势,易感品种‘甬优9号’在受到稻曲菌诱导后表达受到抑制。这些结果表明,OsWRKY71的表达对稻曲菌侵染有应答响应,很可能在水稻防御稻曲菌侵染的机制中发挥作用。

关键词: 水稻, WRKY转录因子, 表达分析, 实时定量PCR

Abstract: Proteins of WRKY family,one of the largest families of transcription factors,play an important role in regulation of plant growth and development.In this study,The full length cDNA sequence of OsWRKY71 gene was cloned by SMART\|RACE\|PCR from rice(Oryza sativa L.),which was named as OsWRKY71(GenBank Accession No.KJ137000). The full length cDNA of OsWRKY71 is 1 245 bp,which includes 1 047 bp complete CDS and encodes 348 amino acid residues with a putative molecular mass of 3828 kDa. The bioinformatics characterization indicated that the protein encoded by OsWRKY71 gene has a typically conserved domain structure of WRKY family,and it shared 69% amino acid sequences similarity with WRKYs of wheat. It shared over 68% amino acid sequences similarity with WRKYs of barley. The results of quantitative real\|time PCR(qRT\|PCR) showed that the highest expression level appeared in flag leaf at heading stage. The lowest expression level appeared in root. The expression of OsWRKY71 was up\|regulated at first and then down\|regulated when the resistant line IR28 was inoculated with Ustilaginoidea virens,and expression was suppressed in the infectious line Yongyou 9. These results implied that OsWRKY71 was a pathogen\|responsive gene and might play an important role in rice pathogen resistance.

Key words: rice, WRKY transcription factor, expression analysis, quantitative real\, time PCR