生物学杂志

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铁皮石斛硫氧还蛋白基因(DoTrxL2)的克隆及表达分析

  

  1. 西南交通大学 生命科学与工程学院, 成都610031
  • 出版日期:2018-10-18 发布日期:2018-10-18
  • 通讯作者: 王万军,博士,教授,主要从事植物发育生物学研究,E-mail: wanjunwang@home.swjtu.edu.cn
  • 作者简介:范静,硕士研究生,主要从事植物发育生物学研究,E-mail: 445817926@qq.com
  • 基金资助:
    国家自然科学基金委员会(31371232)

Cloning and expression analysis of thioredoxin-like 2 gene (DoTrxL2) in Dendrobium officinale Kimura et Migo

  1. School of Life Science and Engineering, Southwest Jiaotong University, Chengdu 610031, China
  • Online:2018-10-18 Published:2018-10-18

摘要: 运用cDNA末端快速扩增技术(RACE)和巢式PCR技术克隆出铁皮石斛硫氧还蛋白超家族基因(Thioredoxin like 2, DoTrxL2)。序列分析表明DoTrxL2基因开放阅读框(ORF)长度为639 bp,编码213个氨基酸,编码产物含有一个硫氧还蛋白家族保守的CXXC功能结构域,通过两个半胱氨酸的双硫键得失电子,起到氧化还原作用。分子进化分析显示,DoTrxL2同小兰屿蝴蝶兰、石刁柏亲缘关系较近。qRT-PCR分析显示,DoTrxL2在铁皮石斛原球茎不同发育时期及不同组织中均有表达差异,其中在原球茎形成早期表达量较高,表明该基因对原球茎形成有着促进作用;而在种子和花中表达量高,表明该基因对植物组织的生殖发育具有重要调控作用。

关键词: 铁皮石斛, 硫氧还蛋白基因, RACE, 实时荧光定量PCR

Abstract: The thioredoxin-like 2 gene (DoTrxL2) in Dendrobium officinale Kimura et Migo was cloned by RACE and nest PCR technologies. The full length of DoTrxL2 contained an open reading frame (ORF) of 639 bp encoding 213 amino acid residues, whose product contained a conserved CXXC domain. Phylogenetic analysis showed that the DoTrxL2 had the closest relationship with monocotyledons, such as Phalaenopsis equestris and Asparagus officinalis. qRT-PCR analysis showed that the expression levels of DoTrxL2 were significantly varied in different developmental stages of protocorm and different tissues, which indicated that the DoTrxL2 plays an important role in the development of protocorm and the morphogical establishment of tissues in plant.

Key words: Dendrobium officinale Kimura et Migo, thioredoxin 2 gene, RACE, qRT-PCR