青绿苔草CbSH4基因克隆及基因编辑靶点筛选

doi:10.11733/j.issn.1007-0435.2026.02.006

草地学报 ›› 2026, Vol. 34 ›› Issue (2): 437-446.DOI: 10.11733/j.issn.1007-0435.2026.02.006

• 研究论文 • 上一篇

青绿苔草CbSH4基因克隆及基因编辑靶点筛选

黄扬洁¹, 滕珂², 范希峰², 岳跃森², 张辉², 温海峰², 尹淑霞¹, 刘凌云²

1. 北京林业大学草业与草原学院, 北京 100083;
2. 北京市农林科学院草业花卉与景观生态研究所, 北京 100097

收稿日期:2025-08-12 修回日期:2025-10-20 发布日期:2026-01-22
通讯作者: 尹淑霞，E-mail:yinsx369@163.com;刘凌云，E-mail:liulingyun01@163.com
作者简介:黄扬洁（2001-），女，汉族，湖南永州人，硕士研究生，主要从事草坪草遗传育种研究，E-mail:Younge0712@163.com；
基金资助:
雄安新区科技创新专项（2022XAGG0100）；北京市农林科学院科技创新专项（KJCX20250917，KJCX20251208）；国家自然科学基金项目（U20A2005，32573613）资助

Molecular Cloning and Gene Editing Targets Screening of CbSH4 in Carex breviculmis

HUANG Yang-jie¹, TENG Ke², FAN Xi-feng², YUE Yue-sen², ZHANG Hui², WEN Hai-feng², YIN Shu-xia¹, LIU Ling-yun²

1. School of Grassland Science, Beijing Forestry University, Beijing 100083, China;
2. Institute of Grassland, Flowers, and Ecology, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China

Received:2025-08-12 Revised:2025-10-20 Published:2026-01-22

摘要/Abstract

摘要： 青绿苔草（Carex breviculmis）是一种节水抗旱的重要乡土物种，具有绿期长、耐阴、结实率高等优点。但在实际采收过程中，种子极易落粒的问题限制了种子机械化收获和大规模生产应用。为探究SH4基因在青绿苔草落粒过程中的功能，进一步利用基因编辑技术创制低落粒新种质，本研究克隆青绿苔草CbSH4基因并进行基因编辑靶点筛选。结果表明，CbSH4编码319个氨基酸残基，其编码产物是亲水性蛋白，与康藏嵩草（Carex littledalei）亲缘关系最近。CbSH4含有1个高度保守的Myb4 DNA结合域，属于典型的MYB转录因子。烟草瞬时表达结果显示，CbSH4定位于细胞核和细胞质。实时荧光定量结果表明，CbSH4在穗中的相对表达量显著高于根和叶，在不同发育时期的离区中均有表达。同时，为了在青绿苔草中实现高效的基因编辑，本研究设计了3个特异性靶点，利用sgRNA体外转录和筛选，获得切割效率58.3%的sgRNA1，可用于后续青绿苔草基因编辑。本研究成功克隆了CbSH4并完成初步功能分析，筛选出了高效基因编辑靶点，为创制低落粒的青绿苔草新种质提供了基础。

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

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 harvesting 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. breviculmis 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 residues， was a hydrophilic protein， and was most closely related to Carex littledalei， containing a highly conserved 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 cytoplasm. 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 completed， and the high-efficiency gene editing targets were screened， which provided a basis for the creation of new germplasm with low seed shattering.

Key words: Carex breviculmis, SH4 gene, Subcellular localization, Expression analysis, Gene target screening, Seed shattering

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.

HUANG Yang-jie, TENG Ke, FAN Xi-feng, YUE Yue-sen, ZHANG Hui, WEN Hai-feng, YIN Shu-xia, LIU Ling-yun. Molecular Cloning and Gene Editing Targets Screening of CbSH4 in Carex breviculmis[J]. Acta Agrestia Sinica, 2026, 34(2): 437-446.

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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

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