辣椒CaTUA基因的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of CaTUA Gene in Capsicum annuum L.

作　　者：郝雪峰[1,2] 张子彤 贾晓宇 曹海艳 雷建军[2] HAO Xuefeng;ZHANG Zitong;JIA Xiaoyu;CAO Haiyan;LEI Jianjun(College of Biological Sciences and Technology,Taiyuan Normal University,Jinzhong 030619,Shanxi,China;College of Horticulture,South China Agricultural University,Guangzhou 510642,Guangdong,China)

机构地区：[1]太原师范学院生物科学与技术学院,山西晋中030619 [2]华南农业大学园艺学院,广东广州510642

出　　处：《中国蔬菜》2025年第4期57-65,共9页China Vegetables

基　　金：山西省自然科学基金(20210302123091,202403021212063)。

摘　　要：辣椒作为常异花授粉作物,在生产实际中利用核不育两用系进行制种是理想手段。α-微管蛋白(α-tubulin)是细胞骨架的主要成分,与植物育性相关。以辣椒核不育两用系AB114为材料,采用扩增片段长度多态性(amplifi ed fragment length polymorphism,AFLP)技术得到蕾期基因表达差异片段,利用cDNA末端快速扩增(rapid-amplifi cation of cDNA ends,RACE)技术克隆AFLP-F-57全长。ORF fi nder分析显示,该基因具有1350 bp的最大开放阅读框,编码450个氨基酸;经Standard Protein BLAST和NCBI Conserved Domain Search鉴定,该基因编码蛋白与烟草、辣椒的α-微管蛋白具有高度相似性,均含有α-tubulin保守结构域,故将其命名为CaTUA基因;RT-PCR分析表明,与可育株相比,不育株中该基因的表达量随花蕾发育呈现下降趋势,于大花蕾时期差异显著;系统进化树分析发现CaTUA蛋白与烟草TUA蛋白序列的进化距离较近。研究初步分析了CaTUA基因的功能,为进一步揭示辣椒核不育与CaTUA的关系提供了理论依据。Capsicum annuum L.is a predominantly cross-pollinated crop.In production,its seeds are often produced using nuclear sterility dual-purpose lines.Alpha tubulin(α-tubulin),a major component of the cytoskeleton,is closely related to plant fertility.Using the male-sterile lines‘AB114’of Capsicum annuum L.as the material,amplified fragment length polymorphism(AFLP)technology was employed to obtain differentially expressed gene fragments during the bud stage.Subsequently,the full-length AFLP-F-57 was cloned using rapid amplification of cDNA ends(RACE).The ORF finder results indicated that the gene had the largest open reading frame of 1350 bp,encoding 450 amino acids.Based on Standard Protein BLAST and NCBI Conserved Domain Search results,the protein encoded by this gene showed high similarity toα-tubulin proteins found in tobacco and pepper,containing an conservedα-tubulin domain.Thus,the gene was named CaTUA.RT-PCR analysis revealed that,compared with the fertile plants,the expression level of the gene in the sterile plants decreased with the development of flower buds and showed a significant difference at the large bud stage.Phylogenetic analysis revealed that the CaTUA protein was closely related to the TUA protein from tobacco in terms of sequence evolution.The study provided a preliminary functional analysis of the CaTUA gene,offering a theoretical basis for further elucidating the relationship between nuclear male sterility in Capsicum annuum L.and the CaTUA genes.

关 键 词：辣椒 核不育两用系 Α-微管蛋白 基因克隆 表达分析 CaTUA基因 

分 类 号：S641.3[农业科学—蔬菜学]