毛薯硒代半胱氨酸甲基转移酶SMT基因的克隆与表达分析
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海南大学热带农林学院,海口 570228

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薯蓣种质资源保护(zzzz002007168)


Cloning and Expression Analysis of a Selenocysteine Methyltransferase SMT Gene from Dioscorea Esculenta (Lour) Brukill
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Institute of Tropical Agriculture and Forestry, Hainan University,Haikou, Hainan570228

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    摘要:

    硒代半胱氨酸甲基转移酶(SMT,Selenocysteine methyltransferase)是植物硒代谢的关键酶,在植物硒积累的过程中发挥着重要作用。为了探究毛薯中SMT基因的功能,本研究以优质富硒毛薯Ds148为研究对象,利用 RT-PCR 技术从中克隆到了毛薯SMT基因,命名为DsSMT。DsSMT(登录号:MH046782)包含 1038 bp完整开放阅读框(OFR),预测编码 345 个氨基酸,理论分子量(Mw)为37.24 kD,等电点(pI)为5.01,编码的蛋白与其他植物的SMT蛋白同源性较高,具有较高的保守性。系统进化树分析显示,毛薯与海枣和菠萝的SMT蛋白的亲缘关系较近,聚为同一类。利用 PSORTⅡ在线软件对 DsSMT 蛋白进行预测,结果表明其可能定位于细胞质中。实时荧光定量PCR分析显示,DsSMT基因在毛薯生长发育的各个时期的根、茎、叶、块茎以及块茎的皮组织中均有所表达,其中在毛薯的根和成熟期表达量最高。本研究从毛薯中克隆出了硒代谢相关酶DsSMT基因的 cDNA序列,并对其进行了生物信息学分析;同时又对DsSMT基因在毛薯块茎形成的不同时期、不同组织中的表达情况进行相对定量的研究和分析,为深入研究DsSMT 基因的调控机理提供分子依据,同时也为毛薯硒吸收、同化的分子机理研究奠定基础。

    Abstract:

    Selenocysteine methyltransferase (SMT) is a key enzyme of selenium metabolism pathway in plants and plays important roles in accumulating for selenium in plants. To investigate the function of SMT in Dioscorea Esculenta (Lour) Brukill, the full-length cDNA was cloned from Dioscorea Esculenta (Lour) Brukill Ds148 using reverse transcription-PCR (RT-PCR) technique. This gene was named DsSMT with a GeneBank accession number MH046782. The full-length open reading frame (OFR) of DsSMT was 1038bp that encoded 345 amino acids. The predicted molecular weight is 37.24 kD with the hypothetical isoelectric point of 5.01. DsSMT was highly conserved and grouped with the homologous proteins from Phoenix dactylifera and Ananas comosus. Wolf Psort protection indicated that DsSMT protein was located in cytoplasm. Real-time quantitative PCR (RT-qPCR) analysis revealed the transcripts of DsSMT in different tissues of Dioscorea Esculenta (Lour) Brukill, including roots, stems, leaves, tubers and the peel of tubers. The levels of DsSMT were different among different tissues, and the level was the highest in roots and mature period during the process of Dioscorea Esculenta (Lour) Brukill development, respectively. Furthermore, the expression level of DsSMT was obviously down-regulated during the enlargemental period, and then increased during the mature period. These results isolated the coding sequence of DsSMT gene and unlocked its expressional profiles, which provided a base in future study of molecular regulation mechanism on selenium metabolism in Dioscorea Esculenta (Lour) Brukill.

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杨亚飞,黎丹,黄东益,等.毛薯硒代半胱氨酸甲基转移酶SMT基因的克隆与表达分析[J].植物遗传资源学报,2019,20(4):1087-1092.

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  • 收稿日期:2018-10-27
  • 最后修改日期:2019-01-09
  • 录用日期:2019-01-15
  • 在线发布日期: 2019-07-16
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