葡萄扩展蛋白家族全基因组分析及VvEXPA27功能研究
Genome-wide Analysis of the Expansin Gene Family in Grape(Vitis vinifera)and Functional Study of VvEXPA27
张建鹏 1尚霄丽1
作者信息
摘要
果粒大小是决定葡萄(Vitis vinifera)外观品质与产量的重要因素之一,细胞膨大对于决定葡萄果实最终大小至关重要.扩展蛋白(expansin,EXP)是细胞壁松弛蛋白,通过破坏细胞壁中纤维素微纤维和相关基质多糖之间的非共价键促进细胞膨大.本研究以'黑比诺PN40024'基因组为参考,基于Pfam数据库采用HMMER对葡萄扩展蛋白家族成员进行全基因组鉴定,剔除冗余序列后,共筛到31个成员.通过生物信息学分析筛选到两个候选基因VvEXPA06和VvEXPA27.RT-qPCR结果显示,VvEXPA06仅在花后20d上调表达,VvEXPA27在花后20和48d均上调表达.将VvEXPA27进行拟南芥(Arabidopsis thaliana)异源表达,并对其转基因植株进行功能验证.结果表明,与对照组相比,过表达VvEXPA27的T3代纯合转基因拟南芥的第3片叶的细胞面积、果荚长度和莲座叶直径均明显增加,表明VvEXPA27在细胞膨大过程中起到积极的作用.本研究为解析葡萄果实细胞膨大的分子机制提供理论参考,为其分子育种提供基因资源.
Abstract
The size of berry is one of the important factors determining the appearance quality and yield of grapes(Vitis vinifera),and cell expansion is crucial for determining the final size of berry.Expansin(EXP)is a cell wall relaxation protein,which promotes cell expansion by breaking the noncovalent bond between cellulose microfibers and related matrix polysaccharides in the cell wall.This study used the'Pinot Noir PN4002'genome as a reference and used HMMER to identify the whole genome of grape expansin family members based on Pfam database.After eliminating redundant sequences,a total of 31 members were screened.Two candidate genes,VvEXPA06 and VvEXPA27,were identified through bioinformatics analysis.The RT-qPCR results showed that VvEXPA06 was only upregulated at 20 d after flowering,while VvEXPA27 was upregulated at both 20 and 48 d after flowering.Subsequently,the VvEXPA27 were subjected to heterologous expression in Arabidopsis thaliana and their transgenic plants were functionally validated.Compared with the vector control(VC),the cell area of the third leaf,silique length,and rosette diameter of T3-generation homozygous transgenic A.thaliana overexpressing(OE)the VvEXPA27 gene showed a significant increase.The results showed that VvEXPA27 played a positive role in cell expansion.This study provides a theoretical basis for analyzing the molecular mechanism of berry cell expansion and genetic resources for molecular breeding.
关键词
葡萄/果实膨大/生物信息学/扩展蛋白Key words
Grape/Berry expansin/Bioinformatics/Expansin引用本文复制引用
基金项目
河南省高等学校重点科研项目(22B210009)
出版年
2024
国家科技期刊平台
NSTL
万方数据