[关键词]
[摘要]
目的 克隆铁皮石斛Dendrobium officinale倍半萜合成酶(sesquiterpene synthase,SES)基因,分析其在铁皮石斛不同组织中的表达差异以及在信号分子茉莉酸甲酯(methyl jasmonate,MeJA)诱导下的表达模式。方法 采用RACE技术克隆铁皮石斛SES(DoSES)cDNA全长,利用相关软件和在线网站对其进行生物信息学分析,并使用实时荧光定量PCR技术分析DoSES的表达模式。结果 获得的DoSES cDNA序列(GenBank登录号为KX278311)全长1 890 bp,含有1个1 659 bp的完整开放阅读框(ORF),编码552个氨基酸的多肽链,与其他科属植物的同源性达到60%左右。DoSES基因在铁皮石斛茎中表达量最高,从高到低依次是叶、花、原球茎、根;且受到MeJA的诱导。结论 克隆了铁皮石斛DoSES基因,为进一步研究调控铁皮石斛萜类代谢奠定了理论基础。
[Key word]
[Abstract]
Objective To clone the full-length cDNA encoding sesquiterpene synthase (SES) gene from Dendrobium officinale (DoSES), then to analyze the expression difference in different tissues and expression patterns of DoSES induced by methyl jasmonate (MeJA). Methods RACE was used to clone the full length cDNA of DoSES. Bioinformatics analysis was carried out by using related software and online website. Then, the expression patterns of DoSES were studied by quantitative real-time PCR (qRT-PCR). Results The DoSES gene was obtained (GenBank accession number KX278311), and the full-length cDNA was 1 890 bp, encoding the protein of 520 amino acids. DoSES had highest homology (≈ 60%) with SES proteins from other plants. Tissue expression analysis showed that DoSES had the highest expression in the stems, followed by leaves, flowers, protocorm and roots. The results of qRT-PCR analysis suggested that DoSES could be induced by MeJA. Conclusion The cDNA encoding DoSES was cloned. The research results provide basic materials for further function alanalysis of this gene in the future.
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[基金项目]
安徽省教育厅自然科学重点项目(KJ2015A005);安徽省大学生创新创业训练计划项目(201610364059)