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以秦艽转录组数据的目标序列为模版,经反转录扩增和RACE等方法,获得一条长1 260 bp的WRKY30转录因子cDNA序列,开放阅读框为975 bp,编码324个氨基酸残基。WRKY转录因子是植物特有的一类大家族,参与植物防卫反应,介导植物应答,在植物生长发育和代谢中起调控作用,参与植物的多种生物学过程,该类家族成员中含有绝对保守的WRKYGQK氨基酸残基,且因此得名。该转录因子含有一个WRKY保守结构域和一个C2HC锌指结构,隶属于WRKY基因家族的第Ⅲ类成员,经序列相似性比对将其命名为GmWRKY30。相对定量2-△△Ct方法分析结果显示,GmWRKY30在秦艽根中的相对表达水平最高,茎中最低,和对照相比有显著性差异。在根中,受花生四烯酸胁迫后GmWRKY30的相对表达水平显著降低,银离子胁迫后表达水平显著升高。这些研究结果说明GmWRKY30基因具有组织表达特异性,对不同诱导子的响应强度有差异,推测其可能参与诱导响应的调控机制不同。
Abstract：The target sequence of transcriptome data of Gentiana macrophylla was used as template, the cDNA sequence of WRKY30 transcription factor was isolated by reverse transcription amplification and RACE, which was 1 260 bp in length, with an open reading frame(ORF) of 975 bp which encoded polypeptide of 324 amino acid. The WRKY protein is a super family of transcription factors unique to plants which participates in plant defense and mediates response, plays a regulatory role in plant growth and metabolism, participates in many biological processes of plants, and was so named for all family members contained a conserved amino acid sequence WRKYGQK. WRKY30 transcription factor contained one conserved one WRKY domain and a specific C2HC zinc finger structure, which belonged to the class Ⅲ member of the WRKY gene family and was named as GmWRKY30 by sequence similarity comparison. The result of the method of 2-△△Ct relative quantitation showed that the relative expression of Gm WRKY30 in root of G. macrophylla was the highest, and the lowest in stem which was significantly different from that of the contrast. In roots, the relative expression level of GmWRKY30 was significantly decreased after arachidonic acid stress, and significantly increased after silver ions stress. These findings suggest that the GmWRKY30 gene has tissue-specific expression and differences response to different elicitors, speculated that GmWRKY30 may participate in the induction of response to different regulatory mechanisms.
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基本信息:
DOI：10.13417/j.gab.037.000366
中图分类号:Q943.2;S567.239
引用信息:
[1]何懿菡,刘玉萍,韩立敏,等.秦艽转录因子WRKY30的克隆及表达模式分析[J].基因组学与应用生物学,2018,37(01):366-372.DOI:10.13417/j.gab.037.000366.
基金信息:
国家自然科学基金项目(No.32170338;No.31670299)资助
2018-01-25
2018-01-25