• 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

木薯MePYL12基因克隆及采后生理性变质过程的表达分析

Cloning and Post-harvest Physiological Deterioration Expression of MePYL12 of Cassava

  • 摘要:
      目的  脱落酸受体PYL家族基因为ABA信号通路的重要成员。研究PYL基因在木薯块根的采后生理性变质(post-harvest physiological deterioration, PPD)和非生物胁迫中的功能,能够为进一步研究ABA信号在木薯抗逆和PPD过程中的功能奠定基础。
      方法  本研究从木薯品种SC124中通过RT-PCR技术克隆得到了MePYL12基因,对它的蛋白质进行相关生信分析,如遗传进化关系、结构域、蛋白质结构预测、理化性质及基因的启动子元件分析,同时对MePYL12基因在相关处理和木薯PPD过程中的表达量进行分析。
      结果  (1)克隆得到的MePYL12长度为567 bp,氨基酸数量为188,理论等电点为5.55,三级结构预测显示含有典型PYL螺旋手柄结构,与蓖麻和橡胶树中PYL蛋白序列的相似性较高,达到了86.77 %和94.68 %。MePYL12基因的蛋白序列含有PYL蛋白家族的保守结构域,特别是ABA结合的区域“Latch”和“Gate”序列100%一致,这些结果表明MePYL12基因编码的蛋白质属于PYL家族成员并且高度保守。(2)10个木薯组织中的MePYL12基因表达分析显示,该基因在分生组织和叶片中的表达水平最高。(3)MePYL12基因主要的启动子元件为光应答元件(Light-responsive motifs)、干旱诱导元件(Drought-induced motif)、ABA应答元件(ABA responsive motif)等元件。(4)MePYL12基因的表达水平显著受到干旱胁迫和ABA处理诱导。在木薯块根的PPD过程也被显著诱导,在6 h达到最高,随后慢慢下降。
      结论  MePYL12基因具有提高植物应对非生物胁迫能力的潜能,同时可能参与了木薯块根的PPD过程,也为后续研究相关功能奠定了基础。

     

    Abstract:
      Objective  Functions of PYL gene in the post-harvest physiological deterioration (PPD) and under abiotic stresses of cassava (Manihot esculenta Crantz) were studied for further research on the key component of the plant abscisic acid (ABA) signaling pathway, PYR/PYL/RCARs family.
      Method  The PYL gene, MePYL12 was cloned from cassava SC 124 by RT-PCR. Bioinformatics was used to analyze the physicochemical properties, conserved domain, genetic evolutionary relationship, protein structure prediction, promoter elements as well as the expressions of MePYL12 in PPD and under abiotic stresses.
      Result  (1) The full-length cDNA of MePYL12 was 567 bp encoded a polypeptide of 188 amino acids with a predicted relative molecular mass of 20.8kD and an isoelectric point of 5.55. The predicted tertiary structure contained a helix. Its multiple protein sequence alignment showed high similarities with the PYL proteins in Hevea brasiliensis (94.68%) and Ricinus communis (86.77 %). MePYL12 consisted of the conserved motifs of the PYL family, such as the ABA binding region "Latch" and "Gate" suggesting it to be a genuine member from the family and highly conserved. (2) The expressions of MePYL12 in 10 different types of cassava tissue/organ were high in the root apical meristem, shoot apical meristem, and leaf. (3) The promoter element analysis showed that the gene contained the light-responsive, drought-induced MBS, and ABA responsive ABRE motifs. (4) The expression of MePYL12 was significantly upregulated by ABA treatment and drought stress as well as during PPD that peaked in 6h followed by downregulation.
      Conclusion  MePYL12 might participate in the PPD process improving the ability of cassava plants in dealing with abiotic stresses. Further study on the functions of MePYL12 in cassava is in order.

     

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