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WRKY转录因子是植物中广泛存在的一种转录因子,参与植物的生物胁迫与非生物胁迫。采用RTPCR克隆黄瓜CsWRKY23基因cDNA序列,对序列进行结构域、基因结构分析,构建系统发育树,通过实时荧光定量PCR(qRT-PCR)分析该基因在黄瓜根、叶组织中的表达。结果表明:CsWRKY23的cDNA开放读码框全长1 431bp,含有5个外显子,4个内含子;编码476个氨基酸,含有两个WRKY结构域,编码蛋白的分子量为52.2kDa、等电点为6.43;系统发育分析表明CsWRKY23与拟南芥AtWRKY33同源。qRTPCR结果表明该基因响应低温胁迫,并在根、叶组织中的表达均上调。本研究为进一步鉴定该基因在非生物胁迫的功能奠定了基础。
WRKY transcription factor is a widespread plant transcription factor, involved in plant biotic and abiotic stress. The cDNA sequence of CsWRKY23 gene was cloned by RTPCR. The sequence and structure of CsWRKY23 gene were analyzed. The phylogenetic tree was constructed. The expression of CsWRKY23 gene in cucumber root and leaf tissue was analyzed by real-time quantitative PCR (qRT-PCR). The results showed that the full length cDNA of CsWRKY23 cDNA was 1 431 bp in length and contained 5 exons and 4 introns. It encoded 476 amino acids and contained two WRKY domains. The molecular weight of the encoded protein was 52.2 kDa and its isoelectric Point was 6.43; Phylogenetic analysis showed that CsWRKY23 was homologous to Arabidopsis AtWRKY33. qRTPCR results showed that the gene was upregulated in response to low temperature stress and in roots and leaves. This study laid the foundation for further identification of the function of this gene in abiotic stress.