小拟南芥激素相关基因及GH3.6的克隆与表达  

Analysis of hormone-related genes of Arabidopsis pumila and cloning and expression of GH3.6 gene

作　　者：黄薇 孙琦 刘芳 金玉环 罗先梅 黄先忠[1] Huang Wei;Sun Qi;Liu Fang;Jin Yuhuan;Luo Xianmei;Huang Xianzhong(Special plant genomics laboratory,College of Life Sciences,Shihezi University,Shihezi,Xinjiang 832003,China)

机构地区：[1]石河子大学生命科学学院特色植物基因组学实验室

出　　处：《石河子大学学报（自然科学版）》2019年第3期323-331,共9页Journal of Shihezi University(Natural Science)

基　　金：国家自然科学基金项目(U1303302)

摘　　要：植物激素是植物体内产生的一些微量的、能调节自身生理过程的有机化合物,生长素是一种重要的植物激素,在植物生长发育的过程中起着重要作用。为了探索激素相关基因在短命植物小拟南芥(Arabidopsis pumila)逆境适应过程中的作用,本研究基于小拟南芥响应高盐胁迫叶片转录组数据库,首先筛选出2156个激素相关基因,包括生长素、脱落酸、乙烯、水杨酸、茉莉酸、油菜素内酯、细胞分裂素及赤霉素相关的基因,其中生长素和脱落酸相关基因所占比例最多并且多为上调表达。进一步通过分层聚类(H-Cluster),K均值聚类(K-means Cluster)和密度聚类(SOM Cluster)分析持续上调的基因,发现一个编码吲哚乙酸酰胺合成酶的基因GRETCHEN HAGEN 3.6(GH3.6)在高盐胁迫过程中明显上调表达。采用RT-PCR克隆小拟南芥Ap GH3.6基因,其开放阅读框长为1839 bp,编码612个氨基酸。系统进化分析表明,Ap GH3.6与山"菜(Eutrema salsugineum)GH3.6进化关系最近,属于同一进化分支。转录组数据分析表明在250 m M Na Cl下,Ap GH3.6持续上调表达;实时荧光定量PCR分析表明Ap GH3.6在小拟南芥各个组织中均有表达,但在花中表达量最高;高盐胁迫表达分析表明,随着胁迫时间增加,Ap GH3.6表达量不断升高。为了进一步研究该基因的功能,构建了植物过量表达载体35S∶Ap GH3.6并转化农杆菌GV3101。本研究为深入分析激素相关基因在小拟南芥响应盐胁迫中的功能机制奠定了基础。Phytohormones are an organic compounds produced within plants that occur in extremely low concentrations.Auxin is an important plant hormone,which plays an important role in plant growth and development.To explore the roles of hormone-related genes in stress adaptation in ephemeral plant Arabidopsis pumila 2156 hormone-related genes were screened based on the transcriptome database of Arabidopsis pumila leaves in response to high salinity.They were divided into auxin,abscisic acid,ethylene,salicylic acid,jasmonic acid,brassinosteroids,cytokinin and gibberellin associated-genes,of which auxin and abscisic acid-related genes account for the most proportion and most were up-regulated.Further analysis of the genes that were continuously up-regulated by H-Cluster.K-means Cluster and SOM Cluster revealed that a gene encoding indole acetic acid-amido synthetase GRETCHEN HAGEN 3.6(GH3.6)was significantly up-regulated during high salt stress.The ApGH 3.6 gene was cloned by RT-PCR and the length of its open reading frame contains is 1839 bp and encodes 612 amino acids.Phylogenetic analysis revealed that ApGH3.6 is closely related to the Eutrema salsugineum GH3.6 and they belong to the same evolutionary branch.Transcriptome data revealed that ApGH 3.6 was continuously upregulated in response to 250 mM NaCl stress.Quantitative real-time PCR analysis showed that ApGH 3.6 was expressed in all the tissues of Arabidopsis pumila and its expression was highest in flowers.To further investigate the function of this gene,a plant overexpression vector 35S∶ApGH3.6 was constructed and transformed into Agrobacterium tumerfaciens strain GV3101.This study is helpful to better understanding the molecular mechanism of hormone-related genes in response to salt stress in Arabidopsis pumila.

关 键 词：小拟南芥 GRETCHEN HAGEN3(GH3) 生长素 基因表达 

分 类 号：Q943.2[生物学—植物学]