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铁皮石斛转录因子基因DoWRKY1的克隆与表达分析

Keywords: 铁皮石斛,WRKY转录因子,基因克隆,转录激活,表达分析

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Abstract:

WRKY转录因子是植物中发现的新型转录调控因子,在调控植物生长发育和代谢等生理过程中发挥重要作用。该研究采用RT-PCR和RACE技术从铁皮石斛Dendrobiumofficinale中克隆得到一个新的WRKY转录因子基因cDNA全长,命名为DoWRKY1,提交GenBank获得登录号KF716131。生物信息学分析表明,该基因全长cDNA为1704bp,其中开放阅读框(ORF)1629bp,编码1条由542个氨基酸组成的多肽,具有2个WRKY保守结构域,属于WRKY家族第Ⅰ类转录因子。酵母单杂交表明,DoWRKY1可以激活下游报告基因(His3,Ade2)的表达,具有转录激活功能,属于转录激活子。半定量RT-PCR显示,DoWRKY1在根、茎、叶和类原球茎中均有表达。qRT-PCR分析表明,DoWRKY1响应茉莉酸甲酯(MeJA)和壳聚糖(chitosan)诱导,分别在2h和1h,表达量达到最高峰。研究结果为进一步研究其在铁皮石斛次生代谢过程的调控功能奠定基础。

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