BrHDA6 mediates nonhistone deacetylation of BrSOT12 to positively regulate downy mildew resistance in Brassica rapa

Abstract

Downy mildew is a major disease that significantly impacts the yield and quality of Brassica rapa. While histone deacetylase (HDAC) family members are implicated in stress responses, their role in regulating downy mildew resistance in B. rapa remains unclear. Herein, we treated the susceptible B. rapa line R32 with Trichostatin A (TSA), a potent HDAC inhibitor. Notably, TSA application significantly enhanced the susceptibility of B. rapa seedlings to downy mildew infection, demonstrating that HDAC plays a crucial role in mediating resistance against this pathogen. Subsequently, we conducted phylogenetic analysis of HDAC family members and performed high-throughput sequencing to assess HDAC gene expression patterns in the resistant (R31) and susceptible (R32) lines following downy mildew inoculation. Notably, the expression of BrHDA6 was significantly higher in the resistant line R31 compared to the susceptible line R32, suggesting its potential role in disease resistance. Using a genetic transformation system, we generated stable transgenic B. rapa plants overexpressing or silenced for BrHDA6. Inoculation with the downy mildew pathogen revealed that BrHDA6 positively regulates disease resistance. Modification omics and parallel reaction monitoring analysis demonstrated that BrHDA6 directly reduces the acetylation level of sulphotransferase 12 (BrSOT12), which likely enhances sulfotransferase activity, consequently boosting salicylic acid production during downy mildew infection. Interaction between BrHDA6 and BrSOT12 was further validated through yeast two-hybrid and dual-luciferase assays. Our study reveals that BrHDA6 confers downy mildew resistance in B. rapa through nonhistone protein deacetylation of BrSOT12, uncovering a novel regulatory mechanism in plant-pathogen interactions.

Acknowledgments

This work was supported by grants from the Beijing Rural Revitalization Agricultural Science and Technology Project [grant number NY2401140000]; the Collaborative Innovation Center of BAAFS [grant number KJCX20240408]; the National Natural Science Foundation of China [grant number 32372688]; the China Agriculture Research System of MOF and MARA [grant number CARS-A03]; the Collaborative Innovation Program of the Beijing Vegetable Research Center [grant number XTCX202302]; and the Foundation for Reform and Development of BVRC [grant number KYCX202302].

Author Contributions

S.C.Y., J.X.W., and M.Z. conceived and designed the experiment. J.X.W. performed the majority of the experiments and data analysis with partial contributions from T.M. M.Z. and B.Z. provided significant assistance with the experimental work. S.C.Y. supervised the study. M.Z. offered ideas to address challenges encountered during the research. J.X.W. drafted the manuscript. All authors have read and approved the final version of the manuscript for publication.

Data availability

Material and data availability All data required for evaluating the conclusions in the paper are available. All materials generated in this study are available from the corresponding author S.C.Y.

Conflict of interest statement

The authors have no conflicts of interest to declare.

Supplementary Data

Supplementary data is available at Horticulture Research online.

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