农业生物技术学报2024，Vol.32Issue(8)：1729-1741,13.DOI:10.3969/j.issn.1674-7968.2024.08.002
水稻TFⅡB相关因子OsBRF1的功能探析
Functional Analysis of Rice(Oryza sativa)TFⅡB-related Factor OsBRF1
摘要
Abstract
In most eukaryotes,transcription factor ⅡB(TFⅡB)is a general transcription factor of RNA polymerase Ⅱ in transcription of DNA into RNA.TFⅡB-related factor 1(BRF1)serves as the general transcription factor for RNA polymerase Ⅲ,responsible for synthesizing tRNA and 5S rRNA.In Arabidopsis thaliana,AtBRF1 and AtBRF2 participate in gametogenesis in a functionally redundant manner.However,the function of BRF1 in rice(Oryza sativa),an important monocotyledonous model plant and staple crop,has not been reported yet.In this study,OsBRF1 gene promoter-driven β-glucuronidase(GUS)expression vector was constructed.The results of GUS staining showed that OsBRF1 exhibited high expression levels in root,leaf,leaf sheath,stigma,mature root,mature leaf,panicle,pollen,endosperm,and anther,but it was not expressed in the glume.The fusion expression vector of OsBRF1 and GFP was transfected into Nicotiana benthamiana leaves via Agrobacterium tumefaciens,and it was found that OsBRF1 protein predominantly localized in the nucleus.Unable to obtain homozygous frameshift mutant plants using CRISPR/Cas9 gene editing methods indicated that OsBRF1 gene was homozygous lethal.RNAi-mediated silencing led to a significant reduction in pollen activity and fertility(P＜0.05),suggesting a potential role of OsBRF1 in regulating rice fertility by modulating pollen viability.Furthermore,yeast two-hybrid and bimolecular fluorescence complementation assays demonstrated interaction between OsBRF1 and OsTBP1～3(TBP:TATA-binding protein),indicating its potential role as a general transcription factor.This study provides basic material for further exploration of the gene function and regulatory mechanisms of OsBRF1.关键词
水稻/TFⅡB(transcription factorⅡB)家族/表达分析/功能分析/蛋白互作Key words
Rice/Transcription factor ⅡB(TFⅡB)family/Expression analysis/Functional analysis/Protein interaction分类
农业科技引用本文复制引用
陈歆惠,陈广娜,宁鹤,陈家洛,王思怡,朱诚,杨素..水稻TFⅡB相关因子OsBRF1的功能探析[J].农业生物技术学报,2024,32(8):1729-1741,13.基金项目
国家重点研发计划(2018YFE0111900) （2018YFE0111900）
国家自然科学基金(32200251) （32200251）
浙江省博士后科研项目择优资助(一等资助)(ZJ2023038) （一等资助）
