研究论文

Fol-milR1调控SlyFRG5甲基化水平介导番茄抗枯萎病免疫应答的功能解析

  • 王月 ,
  • 李彦婷 ,
  • 肖颖 ,
  • 叶红霞 ,
  • 陈舒曼 ,
  • 欧阳寿强
展开
  • 浙江师范大学生命科学学院, 金华 321004

收稿日期: 2025-07-21

  修回日期: 2025-08-19

  网络出版日期: 2025-09-05

基金资助

浙江省自然科学基金(No.KYZ34423025)和国家自然科学基金面上项目(No.31972351)

SlyFRG5 methylation level activated by Fol-milR1 mediates immune response against tomato wilt disease

  • YU Ru ,
  • LI Pan-Ting ,
  • XIAO Ying ,
  • YE Hong-Xia ,
  • CHEN Shu-Man ,
  • OU Yang-ShouJiang
Expand
  • School of Life Sciences, Zhejiang Normal University, Jinhua 321004

Received date: 2025-07-21

  Revised date: 2025-08-19

  Online published: 2025-09-05

摘要

番茄(Solanum lycopersicum)作为一种营养丰富的蔬菜作物, 在全球范围内被广泛种植。由尖孢镰刀菌番茄专化型(Fusarium oxysporum f.sp. lycopersici, Fol)引起的番茄枯萎病是一种土传真菌病害, 给全球番茄农业生产造成了极大的经济损失。Small RNA (sRNA)介导的DNA甲基化(RdDM)植物抗病分子机制研究备受关注, 但关于病原菌sRNA跨界调控宿主DNA甲基化介导番茄抗枯萎病的分子机制尚未见报道。本课题组前期研究结果表明, 尖孢镰刀菌的效应分子Fol-milR1在病原菌侵染过程中跨界转运至番茄体内, 并通过与SlyAGO4a结合挟持番茄免疫体系。在此基础上, 本研究结果进一步表明SlyAGO4a以负调控的方式参与番茄枯萎病的抗性; 利用全基因组甲基化测序及分子生物学手段鉴定到Solyc02g081370 (SlyFRG5)的甲基化水平与Fol-milR1与直接关联, 其甲基化类型以CG为主, 且发生在基因编码区内; SlyFRG5通过调控宿主ROS的积累参与番茄对枯萎病的免疫。综上所述, 本研究解析了Fol-milR1-SlyAGO4a-SlyFRG5功能模块介导番茄抗枯萎病的分子机制, 为探索番茄抗枯萎病品种的培育和品质改良提供新的思路。

本文引用格式

王月 , 李彦婷 , 肖颖 , 叶红霞 , 陈舒曼 , 欧阳寿强 . Fol-milR1调控SlyFRG5甲基化水平介导番茄抗枯萎病免疫应答的功能解析[J]. 植物学报, 0 : 1 -0 . DOI: 10.11983/CBB25129

Abstract

INTRODUCTION: Tomato wilt disease, caused by Fusarium oxysporum f. sp. lycopersici (Fol), is a soil borne fungal disease that causes significant economic losses to global tomato agricultural production. The molecular mechanism of pathogen sRNA cross-border regulation of host DNA methylation mediated tomato resistance to wilt disease is still unclear. Here, we illustrate that SlyFRG5 methylation level activated by Fol-milR1 mediates immune response against tomato wilt disease. 


RATIONALE: The latest research shows that sRNA derived from pathogens can serve as novel effectors for cross-border participation in host-pathogen interactions. Our previous research documented that Fol-milR1 was transferred horizontally into host cells as an effector. Fol-milR1 repressed the expression of a resistant gene SlyFRG4 in tomato, as well as binded to SlyAGO4a leading to interfere with the host immune system to achieve successful infection. Therefore, this research further determines whether transmission of?Fol-milR1 into tomato causes DNA methylation in the tomato genome and elucidate other aspects of the molecular mechanism involving the action of Fol-milR1.


RESULTS: The results indicate that SlyAGO4a negatively participates in the resistance to tomato wilt disease. The methylation level of Solyc02g081370 (SlyFRG5) was identified to be directly associated with Fol-milR1 using whole genome methylation sequencing and molecular biology methods. SlyFRG5 loss-of-function alleles created using CRISPR/Cas9 in resistant cultivar tomato Motelle exhibited enhanced disease susceptibility to Fol, further supporting the idea that SlyFRG4 is essential for tomato wilt disease resistance. SlyFRG5 contributed wilt disease resistance by regulating the accumulation of host ROS in tomato. 


CONCLUSION: Combining previous research, we concluded that,during the infection Fol, Fol-milR1 was transferred into host cells to hijack SlyAGO4a which leads to methylation a Fusarium disease resistant gene SlyFRG5. This study analyzed how the Fol-milR1-SlyAGO4a-SlyFRG5 functional module mediated tomato resistance to wilt disease, which will provide a new ideas for exploring the cultivation and quality improvement of tomato resistance to Fusarium disease.



The Fol-milR1-SlyAGO4a-SlyFRG5 functional module mediated tomato resistance to wilt disease. (A) Distribution of methylation type in the CDS (Partial region) region of SlyFRG5; (B) The transcription level of SlyFRG5 at different time points after pathogen infection in MM; (C) Leaf DAB (H2O2) and NBT (•O₂⁻) staining (bar=1 cm).

参考文献

[1]Agorio A, Vera P(2007).ARGONAUTE4 is required for resistance to Pseudomonas syringae in Arabidopsis.Plant Cell, 19:3778-3790. [2]Baulcombe D(2004).RNA silencing in plants.Nature, 431:356-363. [3]Baxter A, Mittler R, Suzuki N(2014).ROS as key players in plant stress signalling.J Exp Bot., 65:1229-1240. [4]Bhattacharjee S, Zamora A, Azhar MT, Sacco MA, Lambert LH, Moffett P(2009).Virus resistance induced by NB-LRR proteins involves Argonaute4-dependent translational control.Plant J., 58:940-951. [5]Cao D, Ju Z, Gao C, Mei X, Fu D, Zhu H, Luo Y, Zhu B(2014).Genome-wide identification of cytosine-5 DNA methyltransferases and demethylases in Solanum lycopersicum.Gene, 550:230-237. [6]Chan SW, Henderson IR, Jacobsen SE(2005).Gardening the genome: DNA methylation in Arabidopsis thaliana.Nat Rev Genet., 6:351-360. [7]Chan SW, Zilberman D, Xie Z, Johansen LK, Carrington JC, Jacobsen SE(2004).RNA silencing genes control de novo DNA methylation..Science, 303:1336-1336. [8]Di X, Takken FL, Tintor N(2016).Fusarium oxysporum. Front Plant Sci. 7, 170..Front Plant Sci., 16:1-20. [9]Guo X, Xie Q, Li B, Su H(2020).Molecular characterization and transcription analysis of DNA methyltransferase genes in tomato (Solanum lycopersicum). Genet Mol Biol. 43, e20180295..Genet Mol Biol., 43:1-15. [10]Gao Y, Li SJ, Zhang SW, Feng T, Zhang ZY, Luo SJ, Mao HY, Borkovich KA, Ouyang SQ.(2021).SlymiR482e-3p mediates tomato wilt disease by modulating ethylene response pathway.Plant Biotechnol J., 19:17-19. [11]Havecker ER, Wallbridge LM, Hardcastle TJ, Bush MS, Kelly KA, Dunn RM, Schwach F, Doonan JH, Baulcombe DC(2010).The Arabidopsis RNA-directed DNA methylation argonautes functionally diverge based on their expression and interaction with target loci.The Plant cell, 22:321-334. [12]Henderson IR, Jacobsen SE(2007).Epigenetic inheritance in plants.Nature, 447:418-424. [13]Huang W, Xian Z, Hu G, Li Z(2016).SlAGO4A, a core factor of RNA-directed DNA methylation (RdDM) pathway, plays an important role under salt and drought stress in tomato.Mol Breed, 36:1-13. [14]Ji HM, Mao HY, Li SJ, Feng T, Zhang ZY, Cheng L, Luo SJ, Borkovich KA, Ouyang SQ(2021).Fol-milR1,a pathogenicity factor of Fusarium oxysporum,confers tomato wilt disease resistance by impairing host immune responses.New Phytol., 232:705-718. [15]Ji HM, Zhao M, Gao Y, Cao XX, Mao HY, Zhou Y, Fan WY, Borkovich KA, Ouyang SQ, Liu P(2018).FRG3, a Target of slmiR482e-3p, Provides Resistance against the Fungal Pathogen Fusarium oxysporum in Tomato.Front Plant Sci., 9:1-11. [16]Jiang G, Liu D, Yin D, Zhou Z, Shi Y, Li C, Zhu L, Zhai W(2020).A rice NBS-arc gene conferring quantitative resistance to bacterial blight is regulated by a pathogen effector-inducible miRNA.Mol Plant, 13:1752-1767. [17]Law JA, Jacobsen SE(2010a).Establishing,maintaining and modifying DNA methylation patterns in plants and animals.Nat Rev Genet., 11:204-220. [18]Law JA, Jacobsen SE(2010b).Establishing,maintaining and modifying DNA methylation patterns in plants and animals.Nat Rev Genet., 11:204-220. [19]Li C, Schilmiller AL, Liu G, Lee GI, Jayanty S, Sageman C, Vrebalov J, Giovannoni JJ, Yagi K, Kobayashi Y, Howe GA(2005).Role of beta-oxidation in jasmonate biosynthesis and systemic wound signaling in tomato.Plant Cell, 17:971-986. [20]Matzke MA, Mosher RA(2014).RNA-directed DNA methylation: an epigenetic pathway of increasing complexity.Nat Rev Genet., 15:394-408. [21]Ouyang S, Park G, Atamian HS, Han CS, Stajich JE, Kaloshian I, Borkovich KA(2014).MicroRNAs suppress NB domain genes in tomato that confer resistance to Fusarium oxysporum..PLoS Pathog., 10:1-15. [22]Pradhan M, Pandey P, Baldwin IT, Pandey SP(2020).Argonaute4 modulates resistance to Fusarium brachygibbosum infection by regulating Jasmonic Acid signaling.Plant Physiol., 184:1128-1152. [23]Srinivas C, Nirmala Devi D, Narasimha Murthy K, Mohan CD, Lakshmeesha TR, Singh B, Kalagatur NK, Niranjana SR, Hashem A, Alqarawi AA, Tabassum B, Abd Allah EF, Chandra Nayaka S(2019).Fusarium oxysporum fsp. lycopersici causal agent of vascular wilt disease of tomato: Biology to diversity– A review. Saudi J Biol Sci..Saudi J Biol Sci., 26:1315-1324. [24]Wang B, Sun Y, Song N, Zhao M, Liu R, Feng H, Wang X, Kang Z(2017).Puccinia striiformis fsp. tritici microRNA-like RNA 1 (Pst-milR1), an important pathogenicity factor of Pst, impairs wheat resistance to Pst by suppressing the wheat pathogenesis-related 2 gene.New Phytol., 215:338-350. [25]Wang J, Liu X, Zhang A, Ren Y, Wu F, Wang G, Xu Y, Lei C, Zhu S, Pan T, Wang Y, Zhang H, Wang F, Tan YQ, Wang Y, Jin X, Luo S, Zhou C, Zhang X, Liu J, Wang S, Meng L, Wang Y, Chen X, Lin Q, Zhang X, Guo X, Cheng Z, Wang J, Tian Y, Liu S, Jiang L, Wu C, Wang E, Zhou JM, Wang YF, Wang H, Wan J(2019).A cyclic nucleotide-gated channel mediates cytoplasmic calcium elevation and disease resistance in rice.Cell Res., 29:820-831. [26]Wang M, Weiberg A, Jin H(2015).Pathogen small RNAs: a new class of effectors for pathogen attacks.Mol Plant Pathol., 16:219-223. [27]Weiberg A, Wang M, Lin FM, Zhao H, Zhang Z, Kaloshian I, Huang HD, Jin H(2013).Fungal small RNAs suppress plant immunity by hijacking host RNA interference pathways.Science, 342:118-123. [28]Yu A, Lepère G, Jay F, Wang J, Bapaume L, Wang Y, Abraham AL, Penterman J, Fischer RL, Voinnet O, Navarro L(2013).Dynamics and biological relevance of DNA demethylation in Arabidopsis antibacterial defense.Proc Natl Acad Sci U S A, 110:2389-2394. [29]Zilberman D, Cao X, Jacobsen SE(2003).ARGONAUTE4 control of locus-specific siRNA accumulation and DNA and histone methylation.Science, 299:716-719. [30]Zilberman D, Cao X, Johansen LK, Xie Z, Carrington JC, Jacobsen SE(2004).Role of Arabidopsis ARGONAUTE4 in RNA-directed DNA methylation triggered by inverted repeats.Curr Biol., 14:1214-1220.
文章导航

/