白芨鲨烯合酶基因BsSS的克隆和表达分析  

Cloning and Expression Analysis of Squalene Synthase Gene BsSS in Bletilla striata

作　　者：王双双 窦全丽[1] 刘婷 彭宇龙 张柏林[1] 曾伯平[1] Wang Shuangshuang;Dou Quanli;Liu Ting;Peng Yulong;Zhang Bolin;Zeng Boping(College of Biological and Agricultural Science and Technology,Zunyi Normal University,Zunyi,563006)

机构地区：[1]遵义师范学院生物与农业科技学院,遵义563006

出　　处：《分子植物育种》2023年第12期3882-3888,共7页Molecular Plant Breeding

基　　金：贵州省委组织部人才项目赤水河流域环境保护与山地农业发展人才基地开放基金项目;贵州省教育厅专项科研资金项目(黔教合KY字[2012]01712号);遵义师范学院赤水河流域资源保护与开发研究院开放基金项目(遵师CSHKF[2019]06号);遵义市科技计划项目(遵市科合HZ字[2021]196号)共同资助。

摘　　要：鲨烯合酶(SS)在三萜化合物的合成途径中是关键的限速酶。为研究分析白芨鲨烯合酶BsSS的序列信息与表达模式,揭示其基因功能,基于白芨转录组数据,采用RT-PCR技术获得白芨BsSS基因CDS全长,利用生物信息学方法分析BsSS编码蛋白的分子特征,并进行氨基酸多序列比对和进化关系分析,采用实时荧光定量PCR技术检测BsSS基因的组织表达模式。结果表明克隆获得的BsSS基因CDS全长为1230 bp,编码409个氨基酸。BsSS蛋白包含1个跨膜结构域,属于Isoprenoid-Biosyn-C1超家族。与铁皮石斛DcSS和小兰屿蝴蝶兰PeSS基因编码蛋白序列一致性高,并聚为一支。BsSS基因的表达模式具有组织特异性,其转录本在白芨幼苗的根中表达量最高,叶片次之,而在茎和假鳞茎中表达水平最低。对白芨BsSS基因的序列特征与组织表达模式分析为进一步阐明BsSS基因在调控植物萜类物质合成途径中的调控机制提供理论基础。Squalene synthase(SS)is a key rate limiting enzyme in the triterpenoid biosynthesis pathway.To study the sequence information and expression pattern of squalene synthase BsSS in Bletilla striata and reveal its gene function,based on the transcriptome data of B.striata,the full length CDS of BsSS gene was obtained by RT-PCR,the molecular characteristics of BsSS were analyzed by bioinformatics tools.Amino acid sequence alignment and phylogenetic analysis of BsSS were performed.The quantitative real-time PCR(RT-qPCR)was performed to investigate the expression patterns of BsSS.The result showed that the BsSS cDNA contained an open reading frame of 1230 bp nucleotides encoding a 409 amino acids protein.It was predicated that the protein exhibited 1 transmembrane spanning domain belonging to Isoprenoid_Biosym_C1 superfamily.It was highly consistent with DcSS of Dendrobium officinale and PeSS gene of Phalaenopsis paolanthus,and clustered into one branch.The expression of BsSS revealed differential expression by tissue specificity,expression level of BsSS mRNA in root was the highest,followed by leaves,but in stem and pseudobulb were lower than that in other tissues.The analysis of sequence and tissue expression pattern of BsSS gene provided experimental basis for the study of terpenoids biosynthesis in B.striata.

关 键 词：白芨(Bletilla striata) 鲨烯合酶 序列分析 表达分析 

分 类 号：S567.239[农业科学—中草药栽培]