作物学报2024，Vol.50Issue(8)：1971-1988,18.DOI:10.3724/SP.J.1006.2024.31080
小麦泛素结合酶TaUBC16基因的克隆与功能分析
Cloning and functional analysis of ubiquitin-conjugating enzymes TaUBC16 gene in wheat
摘要
Abstract
E2 ubiquitin-conjugating enzyme plays an important role in regulating plant growth and development,and stress signal transduction.In this study,the TaUBC16 gene encoding E2 ubiquitin-conjugating enzyme was cloned from the cDNA of the drought-tolerant wheat cultivar Jinmai 47.The gene was 447 bp in length and encoded 148 amino acids.The cis-acting element analysis showed that the promoter region of TaUBC16 contained various cis-acting elements related to meristem development,stress responses,and plant hormone responses.By the wheat RNA-seq transcriptome data analysis combined with qRT-PCR vali-dation,it was found that TaUBC16 was generally expressed in different tissues/organs and at different growth stages of wheat,whereas the highest expression level was exhibited in developing grains at 30 days after anthesis.The relative expression level of TaUBC16 was highly induced by PEG,mannitol,and ABA stresses.The subcellular localization in tobacco leaves and wheat pro-toplasts showed that TaUBC16 proteins were located in both cytoplasm and nucleus.The phenotypic analysis of the heterologous expression of TaUBC16 in transgenic Arabidopsis revealed that the transgenic lines had earlier flowering time than the wild type,and its seeds were more plumpness with higher 1000-grain weight than the wild type.Based on the polymorphism of the promoter region-388 bp site(T-A),a kompetitive allele-specific PCR(KASP)marker of TaUBC16 gene was developed and its haplotypes were identified.The haplotype TaUBC16-Hap I had higher thousand-kernel weight,kernel length and width than TaUBC16-Hap II,and had been subjected to positive selection in wheat breeding processes in China.The results of this study provide a theoreti-cal basis for further revealing the involvement of the TaUBC16 gene in the regulation of wheat growth and development and the molecular mechanism responding to adverse stresses.关键词
小麦/TaUBC16/基因表达/亚细胞定位/转基因/KASP标记Key words
wheat/TaUBC16/gene expression/subcellular localization/transgenesis/KASP marker引用本文复制引用
高维东,胡城祯,张龙,张艳艳,张沛沛,杨德龙,陈涛..小麦泛素结合酶TaUBC16基因的克隆与功能分析[J].作物学报,2024,50(8):1971-1988,18.基金项目
本研究由中央引导地方科技发展资金项目(23ZYQA0322),甘肃省高校产业支持计划项目(2022CYZC-44),甘肃省重点科技专项(22ZD6NA009),国家自然科学基金项目(32360518,32160487)和甘肃省优秀研究生"创新之星"项目(2023CXZX-684)资助.This study was supported by the Central Guidance on Science&Technology Development of Gansu(23ZYQA0322),the Industrial Support Plan of Colleges and Universities in Gansu Province(2022CYZC-44),the Key Sci&Tech Special Project of Gansu Province(22ZD6NA009),the National Natural Science Foundation of China(32360518,32160487),and the"Innovation Star"Project of Gansu Province Outstanding Postgraduate Students(2023CXZX-684). （23ZYQA0322）
