植物学报 ›› 2024, Vol. 59 ›› Issue (3): 452-462.DOI: 10.11983/CBB23046 cstr: 32102.14.CBB23046
收稿日期:2023-04-07
接受日期:2023-08-07
出版日期:2024-05-10
发布日期:2024-05-10
通讯作者:
孙永伟, 内蒙古大学生命科学学院副教授、硕士生导师。入选内蒙古自治区高等学校青年科技英才计划。先后主持国家自然科学基金项目3项, 内蒙古自然科学基金面上项目3项。主要从事牧草生物学及牧草基因组编辑体系建立相关研究。在Molecular Plant、Plant Biotechnology Journal和Frontiers in Plants Science等期刊发表学术论文10余篇, 授权国家发明专利10余项。E-mail: 基金资助:Received:2023-04-07
Accepted:2023-08-07
Online:2024-05-10
Published:2024-05-10
Contact:
E-mail: 摘要: CRISPR/Cas作为一种新兴的靶向基因组编辑技术, 具有操作过程简便、编辑效率高和支持多靶点编辑等优势, 在植物遗传育种中应用前景广阔。然而对于一些尚未建立遗传转化体系和再生体系的植物, 基因组编辑技术的应用仍然受限。病毒介导的植物基因组编辑技术可不依赖遗传转化和再生等步骤, 即可快速获得无外源转基因成分的基因组编辑植物, 受到广泛关注。该文主要介绍了病毒介导的CRISPR/Cas植物基因组编辑技术的工作原理及优势, 系统总结了该技术在植物基因组编辑领域的应用现状, 并重点讨论了该技术体系存在的问题及挑战, 以期为深入开展这一领域研究提供参考。
胡丹玲, 孙永伟. 病毒介导的植物基因组编辑技术研究进展. 植物学报, 2024, 59(3): 452-462.
Danling Hu, Yongwei Sun. Advances in Virus-mediated Genome Editing Technology in Plants. Chinese Bulletin of Botany, 2024, 59(3): 452-462.
图1 病毒介导的植物基因组编辑技术流程图 (A) 获得可实现基因组编辑的病毒载体; (B) 农杆菌注射法表达植物病毒途径; (C) 无法直接用农杆菌侵染的植物表达病毒途径
Figure 1 Flow chart of virus-mediated genome editing in plants (A) Engineering of viral vectors for transient expression of genome editing reagent; (B) Pathway of using Agrobacterium inoculation method to express plant virus for genome editing; (C) Pathway of delivery of virus to plants which cannot be infected by Agrobacterium inoculation
| 病毒类型 | 病毒名称 | 受体植物 | 递送成分 | 非组织培养稳定遗传效率 | 参考文献 |
|---|---|---|---|---|---|
| 正链RNA病毒 | 烟草脆裂病毒 | 本氏烟草(Nicotiana benthamiana) (Cas9过表达) | sgRNAs | 0.2% | Ali et al., |
| 烟草脆裂病毒和豌豆早枯病毒 | 本氏烟草和拟南芥(Arabidopsis thaliana) (Cas9过表达) | sgRNAs | N.A | Ali et al., | |
| 烟草脆裂病毒 | 本氏烟草(Cas9过表达) | sgRNAs和可移动sgRNAs | 65%-100% | Ellison et al., | |
| 烟草脆裂病毒 | 拟南芥(过表达SunTag系统) | sgRNAs和可移动 sgRNAs | 5%-8% | Ghoshal et al., | |
| 烟草脆裂病毒 | 拟南芥(Cas9过表达) | sgRNAs和可移动sgRNAs | 39%-60% | Nagalakshmi et al., | |
| 烟草脆裂病毒 | 烟草(N. tabacum) (Cas9过表达) | sgRNAs | N.A | Oh et al., | |
| 烟草脆裂病毒和豌豆早枯病毒 | 烟草(Cas9过表达) | sgRNAs | N.A | Kim et al., | |
| 烟草花叶病毒 | 本氏烟草(Cas9过表达) | Cas9和sgRNAs | N.A | Cody et al., | |
| 烟草花叶病毒 | 本氏烟草 | Cas9和sgRNAs | N.A | Chiong et al., | |
| 番茄花叶病毒 | 本氏烟草(分开的SaCas9s部分过表达) | 部分Split-SaCas9s和sgRNAs | N.A | Kaya et al., | |
| 苹果潜隐球形病毒 | 本氏烟草和大豆(Glycine max) (Cas9, Csy4过表达) | sgRNAs | N.A | Luo et al., | |
| 甜菜坏死黄脉病毒 | 本氏烟草(Cas9过表达) | sgRNAs | N.A | Jiang et al., | |
| 大麦条纹花叶病毒 | 本氏烟草、小麦(Triticum aestivum)和玉米(Zea mays) (Cas9过表达) | sgRNAs | N.A | Hu et al., | |
| 大麦条纹花叶病毒 | 小麦(Cas9过表达) | sgRNAs和可移动sgRNAs | 0-17% | Wang et al., | |
| 大麦条纹花叶病毒 | 本氏烟草和小麦(Cas9过表达) | sgRNAs和可移动sgRNAs | 12.9%-100% | Li et al., | |
| 大麦条纹花叶病毒 | 本氏烟草和小麦(Cas9过表达) | sgRNAs和可移动sgRNAs | 0.5%-0.8% | Chen et al., | |
| 大麦条纹花叶病毒 | 本氏烟草和大麦(Hordeum vulgare) (Cas9过表达) | sgRNAs | 100% | Tamilselvan-Nattar- Amutha et al., 2023 | |
| 马铃薯X病毒 | 本氏烟草 | Cas9和sgRNAs | N.A | Ariga et al., | |
| 马铃薯X病毒 | 本氏烟草(Cas9过表达) | sgRNAs和可移动sgRNAs | 22% | Uranga et al., | |
| 狗尾草花叶病毒 | 本氏烟草 | Cas9和sgRNAs | N.A | Zhang et al., | |
| 狗尾草花叶病毒 | 本氏烟草、玉米和狗尾草(Setaria viridis) (Cas9过表达) | sgRNAs | N.A | Mei et al., | |
| 负链RNA 病毒 | 苦苣菜黄网弹状病毒 | 本氏烟草 | Cas9和sgRNAs | N.A | Ma et al., |
| 大麦黄条纹花叶病毒 | 本氏烟草 | Cas9和sgRNAs | N.A | Gao et al., | |
| 番茄斑萎病毒 | 本氏烟草、番茄(Solanum lycopersicum)、辣椒(Capsicum annuum)、灯笼果(Physalis peruviana)和花生(Arachis hypogaea) | Cas9和sgRNAs, Cas12a和crRNA | N.A | Liu et al., | |
| DNA 病毒 | 甘蓝曲叶病毒 | 本氏烟草(Cas9过表达) | sgRNAs | N.A | Yin et al., |
| 棉皱叶病毒 | 拟南芥(Cas9过表达) | sgRNAs和可移动sgRNAs | 4.35%-8.79% | Lei et al., | |
| 棉皱叶病毒 | 棉花(Gossypium spp.) (Cas9过表达) | sgRNAs和可移动sgRNAs | N.A | Lei et al., |
表1 病毒介导的植物CRISPR/Cas基因组编辑技术
Table 1 Virus-mediated CRISPR/Cas genome editing technology in plants
| 病毒类型 | 病毒名称 | 受体植物 | 递送成分 | 非组织培养稳定遗传效率 | 参考文献 |
|---|---|---|---|---|---|
| 正链RNA病毒 | 烟草脆裂病毒 | 本氏烟草(Nicotiana benthamiana) (Cas9过表达) | sgRNAs | 0.2% | Ali et al., |
| 烟草脆裂病毒和豌豆早枯病毒 | 本氏烟草和拟南芥(Arabidopsis thaliana) (Cas9过表达) | sgRNAs | N.A | Ali et al., | |
| 烟草脆裂病毒 | 本氏烟草(Cas9过表达) | sgRNAs和可移动sgRNAs | 65%-100% | Ellison et al., | |
| 烟草脆裂病毒 | 拟南芥(过表达SunTag系统) | sgRNAs和可移动 sgRNAs | 5%-8% | Ghoshal et al., | |
| 烟草脆裂病毒 | 拟南芥(Cas9过表达) | sgRNAs和可移动sgRNAs | 39%-60% | Nagalakshmi et al., | |
| 烟草脆裂病毒 | 烟草(N. tabacum) (Cas9过表达) | sgRNAs | N.A | Oh et al., | |
| 烟草脆裂病毒和豌豆早枯病毒 | 烟草(Cas9过表达) | sgRNAs | N.A | Kim et al., | |
| 烟草花叶病毒 | 本氏烟草(Cas9过表达) | Cas9和sgRNAs | N.A | Cody et al., | |
| 烟草花叶病毒 | 本氏烟草 | Cas9和sgRNAs | N.A | Chiong et al., | |
| 番茄花叶病毒 | 本氏烟草(分开的SaCas9s部分过表达) | 部分Split-SaCas9s和sgRNAs | N.A | Kaya et al., | |
| 苹果潜隐球形病毒 | 本氏烟草和大豆(Glycine max) (Cas9, Csy4过表达) | sgRNAs | N.A | Luo et al., | |
| 甜菜坏死黄脉病毒 | 本氏烟草(Cas9过表达) | sgRNAs | N.A | Jiang et al., | |
| 大麦条纹花叶病毒 | 本氏烟草、小麦(Triticum aestivum)和玉米(Zea mays) (Cas9过表达) | sgRNAs | N.A | Hu et al., | |
| 大麦条纹花叶病毒 | 小麦(Cas9过表达) | sgRNAs和可移动sgRNAs | 0-17% | Wang et al., | |
| 大麦条纹花叶病毒 | 本氏烟草和小麦(Cas9过表达) | sgRNAs和可移动sgRNAs | 12.9%-100% | Li et al., | |
| 大麦条纹花叶病毒 | 本氏烟草和小麦(Cas9过表达) | sgRNAs和可移动sgRNAs | 0.5%-0.8% | Chen et al., | |
| 大麦条纹花叶病毒 | 本氏烟草和大麦(Hordeum vulgare) (Cas9过表达) | sgRNAs | 100% | Tamilselvan-Nattar- Amutha et al., 2023 | |
| 马铃薯X病毒 | 本氏烟草 | Cas9和sgRNAs | N.A | Ariga et al., | |
| 马铃薯X病毒 | 本氏烟草(Cas9过表达) | sgRNAs和可移动sgRNAs | 22% | Uranga et al., | |
| 狗尾草花叶病毒 | 本氏烟草 | Cas9和sgRNAs | N.A | Zhang et al., | |
| 狗尾草花叶病毒 | 本氏烟草、玉米和狗尾草(Setaria viridis) (Cas9过表达) | sgRNAs | N.A | Mei et al., | |
| 负链RNA 病毒 | 苦苣菜黄网弹状病毒 | 本氏烟草 | Cas9和sgRNAs | N.A | Ma et al., |
| 大麦黄条纹花叶病毒 | 本氏烟草 | Cas9和sgRNAs | N.A | Gao et al., | |
| 番茄斑萎病毒 | 本氏烟草、番茄(Solanum lycopersicum)、辣椒(Capsicum annuum)、灯笼果(Physalis peruviana)和花生(Arachis hypogaea) | Cas9和sgRNAs, Cas12a和crRNA | N.A | Liu et al., | |
| DNA 病毒 | 甘蓝曲叶病毒 | 本氏烟草(Cas9过表达) | sgRNAs | N.A | Yin et al., |
| 棉皱叶病毒 | 拟南芥(Cas9过表达) | sgRNAs和可移动sgRNAs | 4.35%-8.79% | Lei et al., | |
| 棉皱叶病毒 | 棉花(Gossypium spp.) (Cas9过表达) | sgRNAs和可移动sgRNAs | N.A | Lei et al., |
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