Journal of Nuclear Agricultural Sciences ›› 2021, Vol. 35 ›› Issue (9): 1994-2001.DOI: 10.11869/j.issn.100-8551.2021.09.1994

• Induced Mutations for Plant Breeding·Agricultural Biotechnology • Previous Articles     Next Articles

Cloning and Expression Analysis of S-adenosylmethionine Synthetase Gene From Carthamus tinctorius L.

TAN Zhengwei1(), LI Lei1, YU Yongliang1, XU Lanjie1, YANG Hongqi1, DONG Wei1, MA Xinming2, LIANG Huizhen1,*()   

  1. 1 Henan Sesame Research Center, Henan Academy of Agricultural Sciences, Zhengzhou, Henan 450002
    2 College of Information and Management Science, Henan Agricultural University, Zhengzhou, Henan 450002
  • Received:2020-05-08 Accepted:2020-07-14 Online:2021-09-10 Published:2021-07-22
  • Contact: LIANG Huizhen

红花S-腺苷甲硫氨酸合成酶基因的克隆与表达分析

谭政委1(), 李磊1, 余永亮1, 许兰杰1, 杨红旗1, 董薇1, 马新明2, 梁慧珍1,*()   

  1. 1 河南省农业科学院芝麻研究中心,河南 郑州 450002
    2 河南农业大学信息与管理科学学院,河南 郑州 450002
  • 通讯作者: 梁慧珍
  • 作者简介:谭政委,男,助理研究员,主要从事分子生药学研究。E-mail: zhwtan@126.com
  • 基金资助:
    国家现代农业产业技术体系,河南省农科院优秀青年基金项目(2020YQ05);国家现代农业产业技术体系,河南省农科院优秀青年基金项目(2020YQ28);河南省博士后基金项目(001803053)

Abstract:

S-adenosylmethionine synthetase is a key enzyme in plant metabolism. In order to explore the mechanism in response of biotic and abiotic stresses, a full cDNA sequence of CtSAMS1 gene was cloned from Carthamus tinctorius L. cultivar Yuhonghua1hao based on the transcriptome data. Meanwhile, the bioinformatics analysis, tissue-specific expression analysis, and expression analysis under different abiotic stresses and hormone treatments were performed. The open reading frame (ORF) of CtSAMS1 gene was 1 173 bp, encoding a protein containing 390 amino acids with a calculated molecular mass (MW) of 42.7 kDa. Protein conserved domain analysis indicated CtSAMS1contained the conserved domain of methionine adenosyltransferase which belonged toS-adenosylmethionine synthase family. The phylogenetic analysis indicated that CtSAMS1 protein had the highest homology with SAMS protein from family Compositae. The results of real-time quantitative PCR showed that the expression of CtSAMS1 gene was the highest in flowers and stems, but very low in other tissues. The transcript level of CtSAMS1 gene increased with the degree of petal senescence. The transcript level of CtSAMS1 was induced by various abiotic stresses including salt, drought and cold. The transcript level of CtSAMS1 also induced by jasmonate (MeJA), salicylic acid (SA) and abscisic acid (ABA), but gibberellin 3 (GA3) can inhibited moderately the transcript ofCtSAMS1. These results provided valuable insights into the resistance mechanism of CtSAMS1 and cultivation of new varieties of Carthamus tinctorius L. stress resistance.

Key words: Carthamus tinctorius L., S-adenosylmethionine synthase, bioinformatics, expression analysis

摘要:

S-腺苷甲硫氨酸合成酶(SAMS)是植物代谢过程中的一个关键酶。为了探究SAMS在红花逆境胁迫方面的作用,本研究根据红花转录组数据从红花品种豫红花1号克隆得到1个SAMS的全长cDNA序列,命名为CtSAMS1,并对其进行生物信息学分析、组织表达特性分析及非生物胁迫和激素诱导表达分析。结果表明,红花CtSAMS1基因的开放阅读框(ORF)长1 173 bp,编码390个氨基酸,其蛋白分子质量为42.7 kDa,蛋白保守区预测表明CtSAMS1具有甲硫氨酸腺苷转移酶的保守结构域,属于S-腺苷甲硫氨酸合成酶家族。系统进化分析显示CtSAMS1与来自菊科的SAMS亲缘关系较近。实时荧光定量PCR(qRT-PCR)检测结果表明,CtSAMS1基因在花和茎中表达量最高,在其他组织部位中表达量极低;CtSAMS1基因表达量随着花瓣衰老程度的增加而升高。盐、干旱和低温胁迫均能够诱导CtSAMS1基因的表达,茉莉酸甲酯、水杨酸、脱落酸能够诱导红花花瓣中CtSAMS1基因表达,赤霉素对CtSAMS1基因表达有一定的抑制作用。本研究为进一步研究CtSAMS1在红花中的抗逆机制,以及为培育红花抗逆新品种奠定了理论基础。

关键词: 红花, S-腺苷甲硫氨酸合成酶, 生物信息学分析, 表达分析