草地学报2026，Vol.34Issue(2)：437-446,10.DOI:10.11733/j.issn.1007-0435.2026.02.006
青绿苔草CbSH4基因克隆及基因编辑靶点筛选
Molecular Cloning and Gene Editing Targets Screening of CbSH4 in Carex breviculmis
摘要
Abstract
Carex breviculamis is an important native species that is water-saving and drought resistant,with advantages such as long green period,shade tolerance,and high fruiting rate.However,in the actual harvest-ing process,the problem of seeds easily falling off limits the mechanization of seed harvesting and large-scale production applications.In order to explore the function of SH4 gene in the process of seed-shattering in C.bre-viculmis and further use gene editing technology to create new germplasm resistant to seed shattering,the CbSH4 gene was cloned and the gene editing target was screened.The CbSH4 encoded 319 amino acid resi-dues,was a hydrophilic protein,and was most closely related to Carex littledalei,containing a highly con-served Myb4 DNA-binding domain,which belonged to a typical MYB transcription factor.The transient expression results of Nicotiana benthamiana showed that CbSH4 protein was localized in the nucleus and cyto-plasm.The real-time fluorescence quantification results showed that the relative expression level of CbSH4 in the spike was significantly higher than that in the roots and leaves,and it was expressed in the abscission zone at different developmental stages.At the same time,in order to achieve efficient gene editing in C.breviculmis,three specific targets were designed in this study,and sgRNA1 was obtained by sgRNA transcription and screening in vitro with a cleavage efficiency of 58.3%,which could be used for subsequent gene editing of C.breviculmis.In this study,CbSH4 was successfully cloned and the preliminary functional analysis was com-pleted,and the high-efficiency gene editing targets were screened,which provided a basis for the creation of new germplasm with low seed shattering.关键词
青绿苔草/SH4基因/亚细胞定位/表达分析/靶点筛选/落粒性Key words
Carex breviculmis/SH4 gene/Subcellular localization/Expression analysis/Gene target screening/Seed shattering分类
农业科技引用本文复制引用
黄扬洁,滕珂,范希峰,岳跃森,张辉,温海峰,尹淑霞,刘凌云..青绿苔草CbSH4基因克隆及基因编辑靶点筛选[J].草地学报,2026,34(2):437-446,10.基金项目
雄安新区科技创新专项(2022XAGG0100) （2022XAGG0100）
北京市农林科学院科技创新专项(KJCX20250917,KJCX20251208) （KJCX20250917,KJCX20251208）
国家自然科学基金项目(U20A2005,32573613)资助 （U20A2005,32573613）
