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

doi:10.11983/CBB25129

摘要/Abstract

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。

关键词: 番茄枯萎病, DNA甲基化, 抗病基因, 抗病分子机制

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).

Key words: Tomato wilt disease, DNA methylation, Disease resistant genes, Molecular mechanism of disease resistance

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. 植物学报, DOI: 10.11983/CBB25129.

Yue Wang, Yanting Li, Ying Xiao, Hongxia Ye, Shuman Chen, Shouqiang Ouyang. SlyFRG5 methylation level activated by Fol-milR1 mediates immune response against tomato wilt disease. Chinese Bulletin of Botany, DOI: 10.11983/CBB25129.

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