植物DNA双链断裂修复机制及其在重离子诱变和基因编辑中的作用

doi:10.13560/j.cnki.biotech.bull.1985.2024-0033

摘要/Abstract

摘要：

在自然界中，植物会遭受各种环境或内源因素导致的DNA损伤，其中DNA双链断裂（double strand breaks, DSBs）的影响最为严重，如果修复不当，将导致基因组不稳定、基因突变甚至细胞死亡。一方面，植物进化出了强大且有序的损伤修复机制，以确保其存活及正常繁衍；另一方面，基于修复过程的容错性及致突变性，T-DNA插入、基因编辑、物理诱变等技术广泛应用于动植物品种改良。相较于哺乳动物，植物DSBs修复通路及其分子机制报道较为有限。本文综述了植物对DSBs损伤的响应、主要修复途径及关键因子，介绍了通路机制尚未完全解析的替代末端连接（alternative end joining, Alt-EJ）的最新研究进展；此外，探讨了重离子束引起的植物DSBs修复特征和多途径选择，以及基于不同DSBs修复途径的基因编辑技术的研究进展，旨在为深入了解植物DSBs损伤响应及修复的分子机制和研发高效生物育种技术提供参考。

关键词: 植物DSBs损伤修复, 同源重组, 非同源末端连接, 重离子束, 基因编辑

Abstract:

Plants are often subjected to DNA damage due to a range of environmental and internal factors in the natural world. Among these factors, DNA double strand breaks（DSBs）have the most serious consequences to plants. If these DSBs are repaired improperly, which may result in genome instability, gene mutations, and even cell death. On one hand, plants have evolved a powerful and well-organized damage repair mechanism to ensure overall survival and normal reproduction. On the other hand, based on the fault-tolerance and mutagenicity of the repair process, a series of techniques such as T-DNA insertion, gene editing, and physical mutagenesis have been widely used for variety improvement in plants and animals. Compared with mammals, reports on DSBs repair pathways and their molecular mechanisms in plants are limited. In this paper, we review the responses of plants to DSBs damage and the main repair mechanisms, and we also focus on the recent advances in alternative end joining（Alt-EJ）, whose mechanism has not yet been fully revealed. In addition, we discuss the characteristics and multi-pathway options of DSBs repair in plants induced by heavy ion beams irradiation, and the progress of gene editing technology based on different DSBs repair pathways. This study aims to provide reference for understanding the molecular mechanism of plant DSBs damage response and developing efficient biological breeding technology.

Key words: damage repair of plant DSBs, homologous recombination, non-homologous end joining, heavy ion beams, gene editing

隆静, 陈婧敏, 刘霄, 张一凡, 周利斌, 杜艳. 植物DNA双链断裂修复机制及其在重离子诱变和基因编辑中的作用[J]. 生物技术通报, 2024, 40(7): 55-67.

LONG Jing, CHEN Jing-min, LIU Xiao, ZHANG Yi-fan, ZHOU Li-bin, DU Yan. Repair Mechanisms of DNA Double-strand Breaks and Their Roles in Heavy Ion Mutagenesis and Gene Editing in Plants[J]. Biotechnology Bulletin, 2024, 40(7): 55-67.

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基因缩写名称 Abbreviation of gene name	英文全称 Full name in English	基因ID Gene ID	功能 Function	参考文献 Reference
MRE11 RAD50 NBS1	meiotic recombination 11 radiation sensitive 50 nijmegen breakage syndrome 1	At5g54260 At2g31970 At3g02680	形成MRE11-RAD50-NBS1复合物行使DNA结合、核酸酶激活和DSBs末端处理	[13⇓-15]
EXO1	exonuclease 1	At1g29630	DSBs末端切除	[16]
RPA1 RPA2 RPA3	replication protein a1 replication protein a2 replication protein a3	At2g06510 At2g24490 At3g52630	ssDNA结合	[17]
IDN2	involved in de novo 2	At3g48670	RPA结合	[17]
RAD51	radiation sensitive 51	At5g20850	链入侵	[18]
RAD52-1 RAD52-1	radiation sensitive 52-1 radiation sensitive 52-2	At1g71310 At5g47870	ssDNA结合和退火	[19]
RAD54	radiation sensitive 54	At3g19210	同源DNA的搜索	[20]
RAD51B RAD51C RAD51D	rad51 paralog b rad51 paralog c rad51 paralog d	At2g28560 At2g45280 At1g07745	ssDNA结合和稳定核蛋白丝	[21]
XRCC2 XRCC3	x-ray repair cross complementing 2 x-ray repair cross complementing 3	At5g64520 At5g54750	ssDNA结合	[21-22]
BRCA1	breast cancer 1	At4g21070	同源DNA的搜索	[23]
BRCA2A BRCA2B	breast cancer 2a breast cancer 2b	At4g00020 At5g01630	同源DNA的搜索	[24]
DMC1	disrupted meiotic cDNA 1	At3g22880	链入侵	[25]
COM1	compositum 1	At3g52115	链交换	[26]
RAD1 RAD10	radiation sensitive 1 radiation sensitive 10	At5g41150 At3g05210	互补链的修剪	[27]
FANCM	fanconi anemia complementation group m	At1g35530	D-Loop形成和单链退火延伸	[28]
PAF1	polymerase associated factor 1	At1g79730	DNA末端结合	[29]
DDRM2	DNA damage response mutants 2	At4g02110	招募RAD51	[30]
SMC5 SMC6A	structural maintenance of chromosomes 5 structural maintenance of chromosomes 6A	At5g15920 At5g07660	促进姐妹染色单体排列	[31]

基因缩写名称 Abbreviation of gene name	英文全称 Full name in English	基因ID Gene ID	功能 Function	参考文献 Reference
MRE11 RAD50 NBS1	meiotic recombination 11 radiation sensitive 50 nijmegen breakage syndrome 1	At5g54260 At2g31970 At3g02680	形成MRE11-RAD50-NBS1复合物行使DNA结合、核酸酶激活和DSBs末端处理	[13⇓-15]
EXO1	exonuclease 1	At1g29630	DSBs末端切除	[16]
RPA1 RPA2 RPA3	replication protein a1 replication protein a2 replication protein a3	At2g06510 At2g24490 At3g52630	ssDNA结合	[17]
IDN2	involved in de novo 2	At3g48670	RPA结合	[17]
RAD51	radiation sensitive 51	At5g20850	链入侵	[18]
RAD52-1 RAD52-1	radiation sensitive 52-1 radiation sensitive 52-2	At1g71310 At5g47870	ssDNA结合和退火	[19]
RAD54	radiation sensitive 54	At3g19210	同源DNA的搜索	[20]
RAD51B RAD51C RAD51D	rad51 paralog b rad51 paralog c rad51 paralog d	At2g28560 At2g45280 At1g07745	ssDNA结合和稳定核蛋白丝	[21]
XRCC2 XRCC3	x-ray repair cross complementing 2 x-ray repair cross complementing 3	At5g64520 At5g54750	ssDNA结合	[21-22]
BRCA1	breast cancer 1	At4g21070	同源DNA的搜索	[23]
BRCA2A BRCA2B	breast cancer 2a breast cancer 2b	At4g00020 At5g01630	同源DNA的搜索	[24]
DMC1	disrupted meiotic cDNA 1	At3g22880	链入侵	[25]
COM1	compositum 1	At3g52115	链交换	[26]
RAD1 RAD10	radiation sensitive 1 radiation sensitive 10	At5g41150 At3g05210	互补链的修剪	[27]
FANCM	fanconi anemia complementation group m	At1g35530	D-Loop形成和单链退火延伸	[28]
PAF1	polymerase associated factor 1	At1g79730	DNA末端结合	[29]
DDRM2	DNA damage response mutants 2	At4g02110	招募RAD51	[30]
SMC5 SMC6A	structural maintenance of chromosomes 5 structural maintenance of chromosomes 6A	At5g15920 At5g07660	促进姐妹染色单体排列	[31]

基因缩写名称 Abbreviation of gene name	英文全称 Full name in English	基因ID Gene ID	功能 Function	参考文献 Reference
Ku70（XRCC6） Ku80（XRCC5）	X-ray repair cross complementing 6 X-ray repair cross complementing 5	At1g16970 At1g48050	结合并保护DSBs末端	[46]
Pol θ	DNA polymerase theta	At4g32700	DNA聚合酶，DSBs末端加工	[47]
MRE11 RAD50 NBS1	Meiotic recombination 11 Radiation sensitive 50 Nijmegen breakage syndrome 1	At5g54260 At2g31970 At3g02680	形成MRE11-RAD50-NBS1复合物行使 DNA结合、核酸酶激活和DSBs末端处理	[13⇓-15]
LIG1	DNA ligase 1	At1g08130	ATP依赖性DNA连接酶	[48]
LIG4	DNA ligase 4	At5g57160	ATP依赖性DNA连接酶	[49]
PARP1 PARP2	Poly（ADP-Ribose）polymerase 1 Poly（ADP-Ribose）polymerase 2	At2g31320 At4g02390	DNA末端结合	[50]
XRCC4	X-ray repair cross complementing 4	At3g23100	结合LIG4	[51]
Pol λ	DNA polymerase lambda	At1g10520	DNA聚合酶，DSBs末端加工	[52]
COM1	Compositum 1	At3g52115	链交换反应	[26]

基因缩写名称 Abbreviation of gene name	英文全称 Full name in English	基因ID Gene ID	功能 Function	参考文献 Reference
Ku70（XRCC6） Ku80（XRCC5）	X-ray repair cross complementing 6 X-ray repair cross complementing 5	At1g16970 At1g48050	结合并保护DSBs末端	[46]
Pol θ	DNA polymerase theta	At4g32700	DNA聚合酶，DSBs末端加工	[47]
MRE11 RAD50 NBS1	Meiotic recombination 11 Radiation sensitive 50 Nijmegen breakage syndrome 1	At5g54260 At2g31970 At3g02680	形成MRE11-RAD50-NBS1复合物行使 DNA结合、核酸酶激活和DSBs末端处理	[13⇓-15]
LIG1	DNA ligase 1	At1g08130	ATP依赖性DNA连接酶	[48]
LIG4	DNA ligase 4	At5g57160	ATP依赖性DNA连接酶	[49]
PARP1 PARP2	Poly（ADP-Ribose）polymerase 1 Poly（ADP-Ribose）polymerase 2	At2g31320 At4g02390	DNA末端结合	[50]
XRCC4	X-ray repair cross complementing 4	At3g23100	结合LIG4	[51]
Pol λ	DNA polymerase lambda	At1g10520	DNA聚合酶，DSBs末端加工	[52]
COM1	Compositum 1	At3g52115	链交换反应	[26]