甘蔗茉莉酸甲基转移酶基因<em>ScJMT</em>的克隆与表达分析

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摘要茉莉酸类物质作为植物体内的內源调节激素,对植物生长发育以及植物对逆境胁迫的响应起着重要作用。茉莉酸甲基转移酶(jasmonic acid carboxyl methyltransferase,JMT)是茉莉酸甲酯合成途径中最后催化茉莉酸合成茉莉酸甲酯的关键酶,控制着植物体内茉莉酸类物质的转化进而影响植物对生物胁迫和非生物胁迫的抵抗能力。本研究基于甘蔗(Saccharum spp. hybrids)转录组数据库,克隆出了甘蔗茉莉酸甲基转移酶ScJMT基因ORF全长序列(GenBank No. MK784565),对该基因进行生物信息学分析并通过qRT-PCR检测不同组织和不同胁迫处理下该基因的表达水平。生物信息学分析结果显示,ScJMT基因开放阅读框为1 110 bp,编码369个氨基酸,等电点为5.28,不稳定系数为56.76,平均疏水性值为-0.135,预测ScJMT基因编码的蛋白为不稳定酸性亲水蛋白。α-螺旋结构和无规则卷曲预测是其二级结构和三级结构主要元件,含有Methyltransf_7超家族的Methyltransf_7保守结构域。系统进化树结果显示,ScJMT与高粱(Sorghum vulgare)和玉米(Zea mays)的甲基转移酶在同一分支上。qRT-PCR结果显示,ScJMT在甘蔗根茎叶中均有表达,且差异不显著。ScJMT在脱落酸(abscisic acid,ABA)、干旱(PEG6000)、高盐(NaCl)、茉莉酸甲酯(methyl jasmonate,MeJA)和粘虫(Mythirnna separata)取食胁迫下表达水平均有显著上升;; 而在MeJA和燕麦食酸菌(Acidovorax avenae subsp. avenae)处理下其表达水平差异没有显著变化。本研究为进一步深入研究该基因的作用提供了基础资料,同时也为培育抗逆的甘蔗新品种提供基因资源资料。

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	陈丽兰
	王亚如
	郭燕芳
	邓祖湖
	王锦达

关键词 ：甘蔗, 茉莉酸甲基转移酶(JMT), 生物信息学, 表达分析

Abstract：Jasmonic acid substances,as endogenous regulatory hormones in plants,play important roles in plant growth and responding to stress. Jasmonic acid carboxyl methyltransferase (JMT) is a key enzyme that catalyzes synthesizing methyl jasmonate (MeJA) from jasmonate. It controls the transformation of jasmonate in plants and affects plants resistance to biological and abiotic stress. Based on sugarcane (Saccharum spp. hybrids) transcriptome database,a full-length of sugarcane jasmonate methyltransferase ScJMT (GenBank No.MK784565) was cloned and analyzed. The expression levels of ScJMT in different organs and expression profiles under different stresses were detected by qRT-PCR. Bioinformatics results showed that the open reading frame (ORF) of ScJMT gene was 1 110 bp encoding 369 aa. The isoelectric point,instability coefficient,and average hydrophobicity value of the ScJMT protein were 5.28,56.76,-0.135,respectively. The ScJMT was an unstable acid hydrophilic protein by prediction. The α-helix structure and random curl constituted its secondary structure and tertiary structure,and it also contained the methylation_7 conserved domain. Through phylogenetic tree analysis showed ScJMT was on the same branch as SvJMT from Sorghum vulgare and ZmJMT from Zea mays. In addition,the spatial expression level of ScJMT was detected and expression profiles under different stress were also determined. There were no significant differences among 3 sugarcane organs,and ScJMT was increased in abscisic acid (ABA),drought (PEG6000),high salinity treatments and oriental armyworm Mythirnna separata feeding stress,while treatments of MeJA and Acidovorax avenae subsp. avenae showed no effect on ScJMT expression. The results make foundations for further research on the role of this gene,and also provide gene resources for the cultivation of new resistant sugarcane varieties.

Key words： Sugarcane (Saccharum spp. hybrids) Jasmonate acid carboxyl methyltransferase (JMT) Bioinformatics Expression analysis

收稿日期: 2020-03-24

ZTFLH:

S33

基金资助:国家重点研发计划(2018YFD201100);国家糖料产业技术体系(CARS-17)

通讯作者: *jdwang@fafu.edu.cn

引用本文:

陈丽兰, 王亚如, 郭燕芳, 邓祖湖, 王锦达. 甘蔗茉莉酸甲基转移酶基因ScJMT的克隆与表达分析[J]. 农业生物技术学报, 2020, 28(11): 1936-1946.
CHEN Li-Lan, WANG Ya-Ru, GUO Yan-Fang, DENG Zu-Hu, WANG Jin-Da. Cloning and Expression Analysis of Jasmonic Acid Carboxyl Methyltransferase Gene (JMT) in Sugarcane (Saccharum spp. hybrids). 农业生物技术学报, 2020, 28(11): 1936-1946.

链接本文:

http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2020/V28/I11/1936

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