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核农学报  2014 

核桃WRKY4基因的克隆与表达分析

DOI: 10.11869/j.issn.100-8551.2014.07.1188, PP. 1188-1196

Keywords: 核桃,WRKY转录因子,基因克隆,实时荧光定量PCR

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Abstract:

WRKY转录调控因子广泛参与植物的生长发育、生物胁迫和非生物胁迫等生理过程。本研究利用reversetranscription-polymerasechainreaction(RT-PCR)及rapidamplificationofcDNAends(RACE)技术克隆了核桃(JuglansregiaL.)WRKY基因cDNA全长序列,命名为WRKY4(GenBank登录号为KC795551.1)。结果表明,克隆到的片段长度为2174bp,完整开放阅读框为1554bp,编码517个氨基酸。序列分析表明WRKY4基因含有2个WRKY保守结构域和2个C2H2锌指结构域,属于WRKY基因家族第Ⅰ类成员。利用实时荧光定量PCR分析4℃低温和自然低温条件对该基因表达的影响以及组织表达模式。结果表明WRKY4受低温诱导表达,能够在低温胁迫的早期响应这一生理过程。WRKY4在树皮(韧皮部)、雌花、花芽和叶片中均有表达,且在树皮中表达量最高,具有组织特异性。研究结果可为核桃抗寒基因工程育种提供基因资源。

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