生物技术通报2026，Vol.42Issue(9)：120-127,8.DOI:10.13560/j.cnki.biotech.bull.1985.2026-0021
拟南芥种子的胚与胚乳中pOp6/LhG4AtO表达系统的构建与验证
Construction and Validation of the pOp6/LhG4AtO Expression System in the Embryo and Endosperm of Arabidopsis Seed
摘要
Abstract
[Objective]To construct the pOp6/LhG4AtO expression system for studying gene function in the embryo and endosperm of Arabidopsis seeds,and to validate its feasibility to provide a reliable tool for tissue-specific gene expression.[Method]Based on the pOp6/LhG4AtO binary transcriptional activation system,previously reported embryo-specific(TWS1)and endosperm-specific(FWA,ESH1)promoters were selected to construct the driver vectors.Meanwhile,a fluorescent reporter gene with enhanced nuclear localization(H2B-Clover)was employed to generate the effector vector.Genetic transformation was performed via Agrobacterium-mediated floral dip method to obtain positive transgenic lines.By crossing the driver lines with the effector line,the tissue-specific expression of the reporter gene was observed in F1 seeds at 2-5 d after pollination,and the expression patterns were compared with publicly available microarray transcriptome data(NCBI GEO;accession number GSE12404).[Result]The pOp6/LhG4AtO expression system was successfully constructed,and the required independent transgenic lines were obtained,including driver lines(pTWS1::LhG4AtO,pFWA::LhG4AtO,pESH1::LhG4AtO)and an effector line(pOp6::H2B-Clover).After crossing the driver line with the effector line,green fluorescence signals driven by pTWS1::LhG4AtO,were specifically and continuously observed at high levels in the nuclei of embryos cells only at 3-5 d after pollination(DAP).Similarly,green fluorescence signals driven by pFWA::LhG4AtO and pESH1::LhG4AtO were observed specifically and continuously at high levels in the nuclei of endosperm cells at 2-5 DAP after pollination.These results demonstrate that the fluorescence expression patterns driven by the pOp6/LhG4AtO system are highly consistent with published microarray transcriptome data.[Conclusion]This study established and validated the pOp6/LhG4AtO system for tissue-specific driver gene expression in the embryo and endosperm of Arabidopsis seeds,providing a reliable tool for functional studies of key genes during seed development.关键词
pOp6/LhG4AtO系统/组织特异性表达/胚/胚乳/种子发育Key words
pOp6/LhG4AtO system/tissue-specific expression/embryo/endosperm/seed development引用本文复制引用
付彩霞,张瑞花,祝云俊,李靖,杨科,王轩鹏..拟南芥种子的胚与胚乳中pOp6/LhG4AtO表达系统的构建与验证[J].生物技术通报,2026,42(9):120-127,8.基金项目
海南大学启动经费项目(XJ2400005259) （XJ2400005259）
