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野菊CibZIP43基因及其启动子的克隆与表达分析
Cloning and Expression Analysis of CibZIP43 Gene and Its Promoter in Chrysanthemum indicium
【摘要】 以野菊(Chrysanthemum indicum)为试材,基于镉胁迫转录组数据,采用RT-PCR克隆、生物信息学分析及启动子功能验证技术,解析CibZIP43基因在镉胁迫响应中的作用机制,以期为发掘野菊抗逆基因资源提供参考依据。结果表明:CibZIP43基因开放阅读框长516 bp,编码171个氨基酸,其蛋白相对分子质量为20 144.29,属不稳定亲水性蛋白,与菊科青蒿同源关系最近;该基因受镉胁迫显著诱导,在叶中的表达量于1 h降至最低,根中于8 h达最低值。启动子序列含MYB结合位点、脱落酸、乙烯及光响应等顺式作用元件。构建5′端缺失片段瞬时表达载体后通过GUS活性检测证实其启动子具有功能活性。
【Abstract】 Chrysanthemum indicum was used as test material to investigate the functional mechanism of CibZIP43 in cadmium stress response through RT-PCR cloning, bioinformatics analysis, and promoter functional verification based on transcriptomic data under cadmium stress, in order to provide reference for the exploration of Chrysanthemum indicium stress resistance gene resources.The results showed that the open reading frame of CibZIP43 gene spanned 516 bp, encoding 171 amino acids with a relative molecular mass of 20 144.29.The encoded protein was identified as an unstable hydrophilic protein, exhibiting closest homology to Artemisia annua.Cadmium stress significantly induced the expression of CibZIP43,with its transcript levels reaching the lowest point in leaves at 1 hour and in roots at 8 hours.The promoter sequence contained multiple cis-acting elements, included MYB-binding sites, abscisic acid, ethylene, and light-responsive elements.Functional activity of the promoter was confirmed by GUS histochemical assay after constructing 5′-end deletion fragments into transient expression vectors.
【Key words】 Chrysanthemum indicium; bZIP; cadmium stress; promoter; expression analysis;
- 【文献出处】 北方园艺 ,Northern Horticulture , 编辑部邮箱 ,2025年11期
- 【分类号】X53;S682.11
- 【下载频次】67