• 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

辣椒海藻糖-6-磷酸合酶基因CaTPS9的克隆及表达分析

Bioinformatics and Expression of CaTPS9 in Chili Peppers

  • 摘要:
      目的  明确辣椒中海藻糖-6-磷酸合酶(TPS)基因CaTPS9的表达特性和生物学功能,进一步了解TPS对辣椒生长中调控非生物胁迫的作用。
      方法  以辣椒品种强丰101为试验材料,克隆CaTPS9基因,并对辣椒CaTPS9的理化性质、蛋白结构、顺式作用元件、系统进化树等进行分析;通过qRT-PCR分析CaTPS9基因在不同组织(商品果果肉、幼果果肉、成熟果果肉、商品果胎座、幼果胎座、成熟果胎座、叶、根、茎、花)和胁迫处理(低温和植物生长调节剂处理)中的表达模式。
      结果  CaTPS9基因CDS序列全长2604 bp,编码867个氨基酸。CaTPS9蛋白包含Glyco_transf_20和Trehalose_PPase两个保守结构域,分子质量为97.60 kDa,不稳定指数为44.27,理论等电点为5.63,亚细胞定位预测CaTPS9蛋白位于细胞质中。生物信息学分析表明,CaTPS9蛋白属于亲水性蛋白,且不存在跨膜结构和信号肽序列,蛋白结构主要由α-螺旋和无规则卷曲组成。系统进化关系分析表明,CaTPS9与烟草(Nicotiana tabacum L.)、番茄(Solanum lycopersicum L.)和马铃薯(Solanum tuberosum L.)中的同源基因亲缘关系较近。启动子顺式作用元件分析表明,CaTPS9启动子区域含有与激素、胁迫及植物生长发育相关的顺式作用元件。此外,CaTPS9在叶片中表达量最高,在商品果胎座中表达量最低。在水杨酸(Salicylic acid,SA) 处理12 h后CaTPS9基因的表达量被显著提升,而低温、吲哚乙酸(3-indoleacetic acid,IAA)、脱落酸(Abscisic acid,ABA)、赤霉素(Gibberellin acid,GA3)和茉莉酸甲酯(Methyl jasmonate,MeJA)处理能够显著抑制CaTPS9基因表达量。
      结论  CaTPS9基因可能通过海藻糖生物合成途径响应逆境胁迫。

     

    Abstract:
      Objective   Bioinformatics and expression of trehalose-6-phosphate synthase (TPS) gene in chili peppers,CaTPS9, were studied to understand its role in response to abiotic stress during the plant growth.
      Method  CaTPS9 was cloned from Qiangfeng 101 to analyze the physicochemical properties, protein structure, cis-regulatory element, and phylogenetic tree. Expressions of the gene in the pericarp and placenta of a commercial pepper as well as the young pericarp, ripened pericarp, young placenta, ripened placenta, leaves, roots, stems, and flowers of chili pepper plants under varied stresses including low temperature, 3-indoleacetic acid(IAA), abscisic acid(ABA), salicylic acid(SA), gibberellin A3(GA3), and methyl jasmonate(MeJA) were determined using qRT PCR.
      Result  The full-length cDNA sequence of CaTPS9 was 2604 bp encoded 867 amino acids. Located in the cytoplasm, the protein contained two conserved domains, Glyco_transf_20 and Trehalose_PPase, with a molecular weight of 97.60 kDa, an instability index of 44.27, and a theoretical isoelectric point of 5.63. It was a hydrophilic protein free of transmembrane structure and signal peptide sequence with a structure consisting mainly of alpha helixes and random coils. Phylogenetically, CaTPS9 was close to the homologous genes in Nicotiana tabacum, Solanum lycopersicum and Solanum tuberosum. It had hormones response, stress response, and plant growth and development cis−elements and was expressed most highly in the leaves, lowly in the placentas of a commercially available pepper, and significantly by 12 h SA induction. Stress, such as low temperature, IAA, ABA, GA3, and MeJA, significantly inhibited the expression.
      Conclusion   It appeared that CaTPS9 might regulate the response of chili pepper plants to various abiotic stresses through the trehalose biosynthetic pathway.

     

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