白及蔗糖合成酶基因的克隆及表达分析  

Cloning and expression analysis of sucrose synthase gene in Bletilla striata

作　　者：蒋素华[1] 牛苏燕 周一冉 崔波[1] 梁芳[1] 袁秀云[1] 马杰[1] JIANG Suhua;NIU Suyan;ZHOU Yiran;CUI Bo;LIANG Fang;YUAN Xiuyun;MA Jie(Bioengineering Research Center,Zhengzhou Normal University,Zhengzhou 450044,China;College of Life Sciences,Zhengzhou University,Zhengzhou 450002,China)

机构地区：[1]郑州师范学院生物工程研究中心,郑州450044 [2]郑州大学生命科学学院,郑州450002

出　　处：《广西植物》2020年第2期192-199,共8页Guihaia

基　　金：河南省重大科技专项(181100110300);河南省产学研合作计划项目(172107000022);河南省科技厅重大专项(701519/702056);郑州师范学院科技创新团队支持计划资助项目~~

摘　　要：为揭示白及蔗糖合成酶基因与生长发育的关系,该研究以白及为材料,利用RT-PCR技术同源克隆白及蔗糖合成酶的关键基因SuSy,对SuSy基因的生物学特性及表达特征进行了分析,并利用实时荧光定量PCR检测SuSy基因在不同组织中的表达规律。结果表明:(1)白及SuSy基因长度为2215 bp,编码737个氨基酸,与铁皮石斛、文心兰和蝴蝶兰的蛋白质氨基酸序列的相似性分别为97%、92%和95%。(2)生物信息学分析表明,SuSy蛋白质序列具有较高的亲水性,与拟南芥SuSy蛋白质氨基酸三级结构一致性为75.2%;系统进化树分析发现,白及SuSy蛋白与铁皮石斛处于同一个分支上。(3)qRT-PCR结果表明,SuSy基因在叶片中的表达量最高,块茎中的表达量最低;成熟叶片的表达量高于未成熟叶片的表达量;数据差异性分析显示,SuSy基因在根、块茎中表达量具有极显著性差异,但在一年生叶和二年生叶中的表达量无显著性差异,幼苗叶和一、二年生叶中表达量具有极显著性差异。由此推测,SuSy基因可能受生长发育的诱导,是调控白及生长发育关键基因。To reveal the function of sucrose synthase(SuSy)gene during the growth and development in Bletilla striata species,the SuSy gene was cloned and its biological and expression characteristics were analyzed.In this study,a key gene(SuSy)for sucrose biosynthesis was homologously cloned from B.striata by RT-PCR technology.Bioinformatics analysis of SuSy gene was performed and real-time fluorescence quantitative PCR was used to detect the expression pattern of SuSy gene in different tissues.The results were as follows:(1)The length of SuSy gene was 2215 bp that encoding 737 amino acids.The similarity of the protein amino acid sequence in Bletilla striata to Dendrobium officinaleis,D.oncidium,and D.phalaenopsis were 97%,92%and 95%,respectively.(2)The results of bioinformatics analysis showed that the SuSy protein sequence was high hydrophily,and the consistency of amino acid tertiary structure of SuSy protein between the Bletilla striata and Arabidopsis thaliana was 75.2%.Phylogenetic tree analysis suggested that the SuSy protein of Bletilla striata and Dendrobium candidum were grouped in the same branch.(3)The qRT-PCR assay showed that the expression level of SuSy gene was the highest in leaves and lowest in tubers.Moreover,the expression of SuSy gene in mature leaves was higher than that in immature leaves.According to the statistical analysis,the expression level of SuSy gene in roots and tubers were very significant,but there were no significant difference in the expression of one-year leaves and two-year leaves,and the expression of seedling leaves were extremely significant.These findings suggested that SuSy gene may be induced by growth and development,and is the key gene for regulation of growth and development in Bletilla striata species.

关 键 词：白及 SUSY 基因 生物信息学分析 表达分析 

分 类 号：Q786[生物学—分子生物学]