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

doi:10.11869/j.issn.100-8551.2021.09.1994

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 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 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

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

TAN Zhengwei, LI Lei, YU Yongliang, XU Lanjie, YANG Hongqi, DONG Wei, MA Xinming, LIANG Huizhen. Cloning and Expression Analysis of S-adenosylmethionine Synthetase Gene From Carthamus tinctorius L.[J]. Journal of Nuclear Agricultural Sciences, 2021, 35(9): 1994-2001.

谭政委, 李磊, 余永亮, 许兰杰, 杨红旗, 董薇, 马新明, 梁慧珍. 红花S-腺苷甲硫氨酸合成酶基因的克隆与表达分析[J]. 核农学报, 2021, 35(9): 1994-2001.

Figures/Tables 8

Table 1 The primer sequences

引物 Primer	序列(5'→3') Squence(5'→3')		用途 Useness
CtSAMS-F	TGTTCACGGCCATCTCACCAAGA		CtSAMS1基因核心片段扩增
CtSAMS-R	CACGATACTCTTGGCGGCTTGCC		CtSAMS1基因核心片段扩增
CtGSP3-1		GACCAAATTGCTGCTGATCTTAA	CtSAMS1基因3'端扩增
CtGSP3-2		CATCTTCCACTTGAACCCATCG	CtSAMS1基因3'端扩增
CtGSP5-1		ATCAGGACGGTGTGAACCCTG	CtSAMS1基因5'端扩增
CtGSP5-2		TTCTGACTTCAGTGAGCTTGGC	CtSAMS1基因5'端扩增
CtSAMS-ORF-F		ATGGAGACCTTCCTATTCACATC	CtSAMS1基因全长扩增
CtSAMS-ORF-R		TCACGCCTTCGGCTTCAGGAC	CtSAMS1基因全长扩增
CtSAMS-qPCR-F		TCCTCATGGAGATGCAGGTC	CtSAMS1基因荧光定量PCR引物
CtSAMS- qPCR-R		CTCAGCCACACCAATAGCGTAC	CtSAMS1基因荧光定量PCR引物
Ct60s-F		GGCAACACTGGTCTTGATCCAT	荧光定量PCR内参引物
Ct60s-R		CTGCGATTGTTGGATAATGGATG	荧光定量PCR内参引物

Fig.1 Amplification of CtSAMS1 from Carthamus tinctorius L. Note: A: Amplification of CtSAMS1 core sequence. B: Amplification of CtSAMS1 3' RACE. C: Amplification of CtSAMS1 5' RACE. D: Amplification of full length of CtSAMS1.

Fig.2 Structural characteristics of CtSAMS1 gene encoding protein Note: A: Predicted secondary structure of CtSAMS1 protein. α: α-helices. β: β-turn. E: Extended strand. R:Random coil. B: The deduced three-dimensional structure of CtSAMS1 protein.

Fig.3 Multiple sequence alignment of CtSAMS1 and SAMS from other plant species Note: At: Arabidopsis thaliana. Mt: Medicago truncatula. Tb: Taraxacum brevicorniculatum. Zm: Zea mays L.

Fig.4 Phylogenetic analysis of CtSAMS1 protein

Fig.5 Relative expression level of CtSAMS1 gene in different tissues and in different developmentalstages of petals of Carthamus tinctorius L.

Fig.6 Relative expression level of CtSAMS1 gene under different abiotic stress

Fig.7 Relative expression level of CtSAMS1 gene under different hormone treatments

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创刊：1987年
主管部门：农业农村部
主办单位：中国原子能农学会与中国农业科学院农产品加工研究所（前中国农业科学院原子能利用研究所）
ISSN：1000-8551

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

红花S-腺苷甲硫氨酸合成酶基因的克隆与表达分析

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