DNA甲基化调控植物组织培养过程的分子机制研究进展

doi:10.12302/j.issn.1000-2006.202302020

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南京林业大学学报（自然科学版） ›› 2023, Vol. 47 ›› Issue (6): 1-8.doi: 10.12302/j.issn.1000-2006.202302020

DNA甲基化调控植物组织培养过程的分子机制研究进展

国颖(), 杨港归(), 吴雨涵, 何杰, 何玉洁, 廖浩然, 薛良交()

林木遗传育种全国重点实验室,南方现代林业协同创新中心,江苏省杨树种质创新与品种改良重点实验室, 南京林业大学林草学院,江苏南京 210037

收稿日期:2023-02-18 修回日期:2023-06-21 出版日期:2023-11-30 发布日期:2023-11-23
通讯作者: *薛良交(lxue@njfu.edu.cn),教授。
基金资助:
国家自然科学基金项目(32171826);江苏省自然科学基金项目(BK20220411)

Recent advances in molecular regulatory mechanisms of DNA methylation in plant tissue culture

GUO Ying(), YANG Ganggui(), WU Yuhan, HE Jie, HE Yujie, LIAO Haoran, XUE Liangjiao()

State Key Laboratory of Tree Genetics and Breeding, Co-Innovation Center for Sustainable Forestry in Southern China, Jiangsu Key Laboratory for Poplar Germplasm Enhancement and Variety Improvement, College of Forestry and Grassland, Nanjing Forestry University, Nanjing 210037, China

Received:2023-02-18 Revised:2023-06-21 Online:2023-11-30 Published:2023-11-23

摘要/Abstract

摘要：

植物细胞具有全能性,创伤和外源激素能够诱导已分化细胞的重编程来再生新的植株,发展的植物组织培养技术已广泛应用于植物快速繁殖、种质保存和性状改良等多个方面。然而,对植物组织培养过程中细胞如何保持分化状态和发育可塑性的分子调控机制仍知之甚少,尤其是在表观遗传学水平上。DNA甲基化是一种进化上保守的表观遗传修饰,能够复杂地协调植物细胞全能性建立和影响其命运转变。在此,以组织培养过程中的愈伤组织形成、体细胞胚发生为切入点,总结了DNA甲基化参与植物再生过程的最新进展。首先,分析了不同植物再生过程中全基因组DNA甲基化变化模式,认为外植体类型和再生阶段均会对DNA甲基化水平产生影响;其次,重点研究了甲基化转移酶(MET1)等在植物再生过程中的作用,以及DNA甲基化调控再生基因表达的分子机制,包括BBM(baby boom),WOX(wuschel-related homeobox),WIN(wound induced dedifferentiation)等基因,最后,讨论了DNA甲基化在植物再生领域的未来研究方向,指出组织培养与基因工程的结合将为农作物和经济、用材林木的高效繁殖和精准培育提供机遇。

关键词: 植物组织培养, DNA甲基化, 愈伤组织, 体细胞胚胎发生

Abstract:

Exerting remarkable cell totipotence, plants are able to regenerate tissues/organs and even individuals from differentiated cells activated by wound stress and/or hormonal cues. Based on the theory of plant cell totipotency, techniques of plant tissue culture have been widely used in rapid propagation, germplasm conservation, and plant breeding as a type of conserved epigenetic modification. However, the understanding of how plant cells retain both differentiated status and developmental plasticity is still obscure, especially at the epigenetic level. DNA methylation is an evolutionarily conserved epigenetic modification that can intricately coordinate cell fate transition and pluripotency establishment during the plant regenerate process. In the work, the recent progress in the regulation of plant regeneration through DNA methylation was summarized, starting from the formation of callus and somatic embryogenesis during tissue culture. Firstly, the change patterns of DNA methylation in different plant regeneration processes were analyzed, showing that both explants type and regeneration phase had an effect on DNA methylation levels. The role of some DNA methyltransferase in plant regeneration was studied, such as DNA Methyltransferase1 (MET1), whose deletion can lead to increased WUS expression and promote shoot regeneration. RNA-directed DNA methylation (RdDM) is the main molecular pathway responsible for de novo DNA methylation in all contexts and is believed to play an important role in plant regeneration. Meanwhile, we analyzed the molecular regulatory mechanisms of DNA methylation on the expression of regenerative genes, such as BBM (baby boom), WOX (wuschel-related homeobox), WIN (wound induced dedifferentiation), etc. Finally, we discussed the future research directions of DNA methylation in the field of plant regeneration. The combination of tissue culture and genetic engineering will provide opportunities for efficient reproduction and precise cultivation of agricultural and forestry crops. Further, the regeneration-related genes reported in this study will provide candidates for plant regeneration research of genetic and molecular mechanisms.

Key words: plant tissue culture, DNA methylation, callus, somatic embryogenesis

中图分类号:

Q943
S722

国颖,杨港归,吴雨涵,等. DNA甲基化调控植物组织培养过程的分子机制研究进展[J]. 南京林业大学学报（自然科学版）, 2023, 47(6): 1-8.

GUO Ying, YANG Ganggui, WU Yuhan, HE Jie, HE Yujie, LIAO Haoran, XUE Liangjiao. Recent advances in molecular regulatory mechanisms of DNA methylation in plant tissue culture[J].Journal of Nanjing Forestry University (Natural Science Edition), 2023, 47(6): 1-8.DOI: 10.12302/j.issn.1000-2006.202302020.

图/表 2

图1

表1

植物组织培养发育过程中DNA甲基化对植物再生关键基因影响"

序号 No.	基因名称 gene symbol	功能 function	物种 species
1	ARR3 (arabidopsis response regulator 3)	参与细胞分裂素调节;5-azaC处理后基因表达上调,发生低甲基化促进桃叶片愈伤组织诱导	桃 Prunus persica^[45]
2	BBM (baby boom)	影响体细胞胚胎发生;表达量升高,甲基化水平降低促进胚胎发生(胚性愈伤组织中高表达)	凹唇姜 Boesenbergia rotunda^[14]
3	CRY1 (cryptochrome 1)	调节细胞分裂素信号,促进芽再生器官的新生	拟南芥 Arabidopsis thaliana^[46]
4	CCD1 (carotenoid cleavage dioxygenases 1)	降解类胡萝卜素;5-azaC处理导致全基因组去甲基化,类胡萝卜素含量降低	柑橘 Citrus paradisi^[47]
5	CMT2/CMT3 (chromomethylase 2/chromomethylase 3)	参与mCHG维持;5-azaC处理抑制了叶外植体愈伤组织的形成和不定芽再生	草莓 Fragaria vesca^[9]
6	CMT3 (chromomethylase 3)	维持DNA甲基化;5-azaC处理后基因表达显著下调,DNA甲基化降低促进桃叶片愈伤组织诱导	桃 P. persica^[45]
		维持DNA甲基化;表达量升高DNA甲基化水平降低,促进体细胞胚胎的发生和再生	凹唇姜 B. rotunda^[48]
7	DRM2 (domains rearranged methyltransferase)	维持CHH甲基化	毛果杨 Populus trichocarpa^[49]
		表达量升高DNA甲基化水平降低,促进体细胞胚胎的发生和再生	凹唇姜 B. rotunda^[48]
8		维持CG甲基化;低甲基化,met1-3突变体芽再生能力更高	拟南芥 A. thaliana^[46]
	MET1 (methyltransferase 1)	维持DNA甲基化;表达量升高DNA甲基化水平降低,促进体细胞胚胎的发生和再生	凹唇姜 B. rotunda^[48]
		维持DNA甲基化;幼苗和嫩叶中偏好表达	柑橘 C. paradisi^[47]
9	ROS1 (repressor of silencing 1)	DNA甲基化水平降低,促进球状胚形成	龙眼 Dimocarpus longan^[30]
10	SERK (somatic embryogenesis receptor-like kinase)	影响体细胞胚胎发生;表达量升高,甲基化水平降低促进胚胎发生(胚性愈伤组织中高表达)	凹唇姜 B. rotunda^[14]
11	WIN (wound-induced)	诱导细胞去分化和增殖;发生去甲基化,基因表达上调促进愈伤组织形成	草莓 F. nilgerrensis^[50]
12	WOX (wuschel-related homeobox)	参与顶端分生组织发生;发生去甲基化,基因表达上调促进愈伤组织形成	草莓 F. nilgerrensis^[50]
13	WUS (wuschel-related homeobox)	调控植物再生;低甲基化激活了生长素和WUS相关基因表达,提高植物再生能力	棉花 Gossypium hirsutum^[51]
		影响体细胞胚胎发生;表达量升高,甲基化水平降低促进胚胎发生(分生组织中表达最高,其次是胚性愈伤)	凹唇姜 B. rotunda^[14]

表1

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DNA甲基化调控植物组织培养过程的分子机制研究进展

Recent advances in molecular regulatory mechanisms of DNA methylation in plant tissue culture

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