茶树WRKY转录因子基因CsWRKY57的克隆及表达分析

摘要/Abstract

摘要： WRKY转录因子是植物特有的一类转录因子,在植物生长发育及胁迫应答过程中均发挥重要的调控作用。为探究WRKY转录因子与茶树抗旱及耐盐性的关系,本研究基于茶树转录组数据库中的检索结果,以陕茶1号1年生茶树为试验材料,克隆获得了1个WRKY转录因子基因,命名为CsWRKY57。生物信息学分析表明,CsWRKY57基因cDNA全长为1β222βbp,编码303个氨基酸,预测分子量为33.5βkD,理论等电点为5.49;另外,蛋白比对分析显示,CsWRKY57包含1个典型的WRKY核心序列和1个C2H2型锌指结构,属于WRKYIIc家族。实时荧光定量PCR分析结果显示,CsWRKY57基因在高盐、干旱、ABA胁迫下均被诱导表达,且表现出先增加后降低的趋势,表明CsWRKY57基因参与了茶树体内干旱、高盐和ABA的调控途径。转录激活活性试验表明,CsWRKY57无转录激活活性,意味着CsWRKY57可能需要与其他元件结合才能启动基因的表达。

关键词: 茶树, CsWRKY57, 克隆, 表达分析

Abstract: The WRKY is one of the characteristic transcription factors in plants, which play important roles in plant growth, development and stress regulation. In order to study the relationship between WRKY transcription factors and stress tolerance of tea plant (Camellia sinensis), a WRKY transcription factor was cloned from tea cultivar ‘Shanchayihao’ and named CsWRKY57, based on the searching result of tea plant transcriptome database. Bioinformatics analysis showed that the full-length sequences of CsWRKY57 was 1β222βbp encoding 303 amino acids. The molecular weight of CsWRKY57 was 33.5βkD and theoretical isoelectric point was 5.49. The BLAST results showed that CsWRKY57 contained one typical WRKY domain and one zinc finger motif (C2H2), suggesting that it was a member of the WRKYIIc family. In addition, quantitative real-time PCR (qRT-PCR) analysis showed that the expression of CsWRKY57 was induced by salt, drought and ABA stresses, and showed a tendency to increase first and then decrease, which implies that CsWRKY57 is involved in the process of tea plant responses to salt, drought and ABA. Furthermore, transcriptional activation activity assays indicated that CsWRKY57 didn't have transcriptional activation activity, which means that CsWRKY57 may be needed to combine with other elements to activate gene expression.

Key words: tea plant (Camellia sinensis), CsWRKY57, cloning, expression analysis

中图分类号:

S571.1
Q52

郭俊红, 王伟东, 谷星, 郭莎莎, 高岳芳, 杨亚军, 肖斌. 茶树WRKY转录因子基因CsWRKY57的克隆及表达分析[J]. 茶叶科学, 2017, 37(4): 411-419.

GUO Junhong, WANG Weidong, GU Xing, GUO Shasha, GAO Yuefang, YANG Yajun, XIAO Bin. Cloning and Expression Analysis of WRKY Transcription Factor Gene CsWRKY57 in Tea Plant (Camellia sinensis)[J]. Journal of Tea Science, 2017, 37(4): 411-419.

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