• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

铁皮石斛DobHLH63基因的克隆及表达分析

Cloning and Expression of DobHLH63 Gene in Dendrobium officinale

  • 摘要:
    目的 碱性螺旋-环-螺旋(basic helix-loop-helix, bHLH)蛋白是真核生物中一类重要的转录因子,在植物生长发育、应答胁迫过程中发挥重要作用。本研究旨在通过克隆铁皮石斛(Dendrobium officinale)DobHLH转录因子,预测该蛋白的结构和功能,并分析该基因在不同条件下的表达模式,为系统阐明该基因的功能奠定基础。
    方法 通过RT-PCR技术克隆DobHLH63基因,并运用生物信息学方法对其进行基因和蛋白水平分析。通过荧光定量PCR技术检测DobHLH63基因对多种植物生长调节剂(如脱落酸、水杨酸、茉莉酸甲酯)以及非生物胁迫(如干旱、盐碱)的响应。
    结果 DobHLH63基因编码区长度为966 bp,编码321个氨基酸,属于植物bHLH蛋白家族;无明显信号肽和跨膜结构域,可能在细胞核和细胞质中发挥作用,有29个磷酸化位点;DobHLH63与紫金龙舌兰(Platanthera zijinensis)中的同源基因PzbHLH63及同基因家族的DobHLH74的亲缘关系最近,推测功能相似;WD40转录因子可能是其重要的互作的蛋白。荧光定量PCR结果表明,DobHLH63基因在叶片中的表达量大于茎,该基因的表达能够响应多种非生物胁迫及植物生长调节剂,尤其是碱性盐胁迫和水杨酸。
    结论 DobHLH63基因可能通过与WD40等转录因子结合调控铁皮石斛对非生物胁迫及植物生长调节剂的响应过程。

     

    Abstract:
    Objective Structure, function, and expression of the transcription factor in the eukaryotes crucial in plant growth, development, and stress response of the basic helix loop helix (bHLH) proteins in Dendrobium officinale were analyzed.
    Methods DobHLH63 was cloned using RT-PCR and bioinformatically analyzed. Spatiotemporal expressions of the gene under various conditions were determined by qRT-PCR.
    Results The coding region of DobHLH63 was 966 bp long and encoded 321 amino acids. The gene was possibly located in the nucleus and cytoplasm with 29 phosphorylation sites but had no apparent signal peptide or transmembrane domain. It closely related to and shared functions with PzbHLH63 and paralog DobHLH74 of Aponogeton madagascariensis. The WD40 transcription factor appeared to be an important interacting protein. The higher expression in the leaves than in the stems responded to abiotic stress and hormone treatments, especially alkaline salt stress and salicylic acid treatment.
    Conclusion DobHLH63 regulated the response of D. officinale to abiotic stress and hormones by binding to the transcription factors, such as WD40.

     

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