紫色马铃薯StCHI基因的克隆与表达分析  

Cloning and Expression Analysis of StCHI Gene in Purple Potato

作　　者：马心萍 朱迪 贺苗苗 MA Xin-ping;ZHU Di;HE Miao-miao(Academy of Agriculture and Forestry Sciences,Qinghai University,Xining 810016,China;Key Laboratory of Qinghai-Tibetan Plateau Biotechnology of Ministry of Education,Xining 810016,China;Key Laboratory of Potato Breeding of Qinghai,Engineering Research Center of Potato in Northwest Region Ministry of Education,Xining 810016,China)

机构地区：[1]青海大学/青海省农林科学院,青海西宁810016 [2]青藏高原生物技术教育部重点实验室,青海西宁810016 [3]青海省马铃薯育种重点实验室/西北马铃薯教育部工程研究中心,青海西宁810016

出　　处：《江西农业学报》2023年第3期12-17,24,共7页Acta Agriculturae Jiangxi

基　　金：青海省科技厅基础研究计划项目(2022-ZJ-712);国家马铃薯产业技术体系(CARS-09)。

摘　　要：查尔酮异构酶是花青素生物合成上游途径的一个关键酶。为研究马铃薯StCHI基因的结构及功能,基于紫色马铃薯品系‘15-12-16’和白色马铃薯品种“下寨65”的转录组数据,利用RT-PCR技术在马铃薯品系“15-12-16”中克隆得到StCHI,并进行了生物信息学分析。利用qRT-PCR技术分析StCHI基因在“15-12-16”不同组织根、茎、叶、花瓣、薯皮、薯肉中的表达水平。结果表明:StCHI基因的CDS为633 bp,该基因编码210个氨基酸,蛋白分子量为23.852 kDa,理论等电点(pI)为4.75,脂溶性指数为91.81,为不稳定的亲水性蛋白。亚细胞定位预测该基因在叶绿体中。蛋白信号肽预测StCHI基因编码的蛋白质为非分泌蛋白,无信号肽,无切割位点。通过同源蛋白多序列比对及进化树分析发现,该基因与风铃辣椒、黄辣椒、黄果枸杞等同一茄科的物种亲缘关系较近,相似度分别为92.31%、91.83%、90.43%。通过实时荧光定量分析发现,该基因在“15-12-16”的各组织中都有所表达,并且该基因具有组织特异性,在花瓣中的表达量较高。为深入研究马铃薯的花青素生物合成提供坚实的理论基础,同时为培育彩色马铃薯以及提取天然色素提供参考价值。。Chalcone isomerase is a key enzyme in the upstream pathway of anthocyanin biosynthesis.In order to study the structure and function of potato StCHI gene,based on the transcriptome data of purple potato line‘15-12-16’and white potato‘Xiazhai 65’,the StCHI gene was cloned from‘15-12-16’by RT-PCR,and bioinformatics analysis was carried out.The expression level of StCHI gene in roots,stems,leaves,petals,skin and tuber of‘15-12-16’was analyzed by qRT-PCR.The results showed that the CDS of StCHI gene was 633 bp,the gene encoded 210 amino acids,the molecular weight of the protein was 23.852 kDa,the theoretical isoelectric point(pI)was 4.75,the fat-solubility index was 91.81,and it was an unstable hydrophilic protein.Subcellular localization predicts that the gene is in chloroplast.Protein signal peptide predicts that the protein encoded by StCHI gene is non-secretory protein,without signal peptide and cutting site.Through homologous protein multiple sequence alignment and phylogenetic tree analysis,it was found that the gene was closely related Capsicum baccatum,Capsicum chinense,Lycium barbarum var.auranticarpum,which were the same species of Solanaceae,with the similarity of 96.67%,92.31%and 91.83%,respectively.Through real-time fluorescence quantitative analysis,it was found that the gene was expressed in all tissues of“15-12-16”,indicated that the gene had tissue specific expression and was highly expressed in petals.This study provides a solid theoretical basis for the in-depth study of anthocyanin biosynthesis in potatoes,provides reference value for cultivating colored potatoes and extracting natural pigments.

关 键 词：马铃薯 花青素 StCHI 表达载体 表达分析 

分 类 号：S532[农业科学—作物学]