苎麻谷胱甘肽-S-转移酶基因BnGSTU1的克隆和表达分析
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宜春学院 生命科学与资源环境学院,江西 宜春 336000,宜春学院 生命科学与资源环境学院,江西 宜春 336000

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江西省教育厅科学技术研究项目(GJJ161008)


Cloning and Expression Analysis of a Glutathione-S-Transferase Gene BnGSTU1 from Ramie (Boehmeria nivea L.)
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College of Life Science and Resources and Environment, Yichun University, Yichun Jiangxi 336000,College of Life Science and Resources and Environment, Yichun University, Yichun Jiangxi 336000

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Science and technology research project of Jiangxi department of education (GJJ161008)

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    摘要:

    谷胱甘肽-S-转移酶(Glutathione S-transferases, GSTs)是一类存在于动植物中的多功能蛋白,对植物抵御逆境胁迫和解除细胞毒素具有重要作用。为探讨GST基因在苎麻耐镉机制中的作用,本研究在苎麻镉胁迫转录组的基础上,采用RACE方法从苎麻(Boehmeria nivea L.)品种中苎一号中克隆到BnGSTU1基因(GenBank登录号为MG941011)的全长cDNA序列,该基因开放读码框为636 bp,编码211个氨基酸,蛋白质分子量为24.07 kD,等电点为5.29。保守结构域分析表明,BnGSTU1蛋白具有GSTs所特有的N端和C端结构域,并含有典型的G位点和H位点,属于Tau类GST亚家族。系统进化树分析表明,BnGSTU1蛋白与拟南芥、橡胶树等物种Tau类GST蛋白的亲缘关系最近。组织表达特异性分析表明,BnGSTU1在苎麻的叶片中表达量最高,其次为茎,根中表达量最低,存在明显的组织特异性。胁迫诱导表达分析表明,BnGSTU1的表达显著受镉的诱导,且表达量随着镉浓度的升高表现出不断增加的趋势。以上结果表明BnMYB1是一个镉应答基因,可能在植物对镉胁迫的适应中发挥重要作用。

    Abstract:

    Glutathione S-transferases (GSTs) are superfamily of multifunctional enzymes which play important roles in stress tolerance and cellular detoxification. In this paper, a GST gene named BnGSTU1 (GenBank accession: MG941011) was isolated from ramie (Boehmeria nivea L.) by amplification of cDNA ends (RACE) based on the analysis of expression profiling of cadmium response genes in ramie. The open reading frame of BnGSTU1 is 636 bp, which encodes 211 amino acid residues. The predicted protein was 24.07 kD with isoelectic point of 5.29. Conserved domain search analysis revealed that BnGSTU1 protein contained a N-teminal domain and a C-teminal domain, as well as glutathione binding sites G and substrate binding pockets H, which showed typical characteristics of Tau class GSTs. Phylogenetic tree analysis showed that BnGSTU1 was closely to Tau class GST in Arabidopsis thaliana, Hevea brasiliensis and so on. Real-time quantitative PCR analysis indicated that BnGSTU1 expressed in root, stem and leaf of ramie, but the expression level of BnGSTU1 in leaf was significantly higher than that in roots. Furthermore, BnGSTU1 gene was significantly induced by cadmium stress, and the expression of BnGSTU1 increased along with Cd2+ concentration. Collectively, the results suggest that BnGSTU1 is a cadmium-responsive factor and may play potential roles in the plant adaption to cadmium stress.

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朱守晶,史文娟.苎麻谷胱甘肽-S-转移酶基因BnGSTU1的克隆和表达分析[J].植物遗传资源学报,2018,19(6):1197-1204.

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  • 收稿日期:2018-02-28
  • 最后修改日期:2018-06-15
  • 录用日期:2018-04-09
  • 在线发布日期: 2018-11-14
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