CsNAC17 enhances resistance to Colletotrichum gloeosporioides by interacting with CsbHLH62 in Camellia sinensis

Abstract

The pathogen Colletotrichum gloeosporioides causes anthracnose, a serious threat to tea trees around the world, particularly in warm and humid regions. RNA-Seq data have previously indicated NAC transcription factors are involved in anthracnose resistance, but underlying mechanisms remain unclear. The BiFC, Split-LUC, and Co-IP assays validated the interaction between CsbHLH62 and CsNAC17 identified through yeast two-hybrid (Y2H) screening. CsNAC17 or CsbHLH62 overexpression enhanced anthracnose resistance, as well as enhanced levels of H₂O₂, hypersensitivity, and cell death in Nicotiana benthamiana. The NBS-LRR gene CsRPM1 is regulated by CsNAC17 by binding directly to its promoter (i.e. CACG, CATGTG), while CsbHLH62 facilitates CsNAC17’s binding and increases transcriptional activity of CsRPM1. Additionally, transient silencing of CsNAC17 and CsbHLH62 in tea plant leaves using the virus-induced gene silencing (VIGS) system resulted in decreased resistance to anthracnose. Conversely, transient overexpression of CsNAC17 and CsbHLH62 in tea leaves significantly enhanced the resistance against anthracnose. Based on these results, it appears that CsbHLH62 facilitates the activity of CsNAC17 on CsRPM1, contributing to increased anthracnose resistance.

Acknowledgements

This work was supported financially by Key Research and Development Program of Jiangsu (BE2023364), National Key Research and Development Program of China (2022YFD1200505), Science and Technology Projects of Nanjing (202210013), Development of New Products from Summer and Autumn Tea in Wen County (2023), National Natural Science Foundation of China (32172628), Nanjing Agricultural Major Technology Collaborative Promotion Plan Project (2024NJXTTG 10) and Research and Demonstration Project of key technologies of tea garden photovoltaic power generation (HNKJ22- H135).

We extend our thanks to Dr Yuehua Ma (Central laboratory of College of Horticulture, Nanjing Agricultural University) for using their microscope (Olympus IX73, Japan); ZEISS LSM 800, Germany; Princeton PIXIS 1024B, USA, and Quantitative real-time PCR (CFX96，Bio-Rad，USA).

Author contributions

R.H. conducted the experiments. R.H., Y.Z., H.M., and A.J. analyzed the data. R.H. wrote the manuscript. S.L., C.M., Y.W., J.Z., Q.H., X.C., and X.L. revised the manuscript. X.L. and S.L. supervised the project. X.L. conceived the project. X.L. acquired funding.

Data availability

All relevant data are included in the article and its supporting materials.

Conflict of interest statement

The authors declare no competing financial interest.

Supplementary Data

Supplementary data is available at Horticulture Research online.

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