CaREM1.4 interacts with CaRIN4 to regulate Ralstonia solanacearum tolerance by triggering cell death in pepper

Abstract

Remorins, plant-specific proteins, have a significant role in conferring on plants the ability to adapt to adverse environments. However, the precise function of remorins in resistance to biological stress remains largely unknown. Eighteen CaREM genes were identified in pepper genome sequences based on the C-terminal conserved domain that is specific to remorin proteins in this research. Phylogenetic relations, chromosomal localization, motif, gene structures, and promoter regions of these remorins were analyzed and a remorin gene, CaREM1.4, was cloned for further study. The transcription of CaREM1.4 in pepper was induced by infection with Ralstonia solanacearum. Knocking down CaREM1.4 in pepper using virus-induced gene silencing (VIGS) technologies reduced the resistance of pepper plants to R. solanacearum and downregulated the expression of immunity-associated genes. Conversely, transient overexpression of CaREM1.4 in pepper and Nicotiana benthamiana plants triggered hypersensitive response-mediated cell death and upregulated expression of defense-related genes. In addition, CaRIN4-12, which interacted with CaREM1.4 at the plasma membrane and cell nucleus, was knocked down with VIGS, decreasing the susceptibility of Capsicum annuum to R. solanacearum. Furthermore, CaREM1.4 reduced ROS production by interacting with CaRIN4-12 upon co-injection in pepper. Taken together, our findings suggest that CaREM1.4 may function as a positive regulator of the hypersensitive response, and it interacts with CaRIN4-12, which negatively regulates plant immune responses of pepper to R. solanacearum. Our study provides new evidence for comprehending the molecular regulatory network of plant cell death.

Acknowledgements

We are grateful to the Natural Science Basic Research Program of Shaanxi Province-Key Project (2023-JC-ZD-12), the National Key R&D Program of China (2021YFD1401000), the Shaanxi Innovation Team Project (2018TD-004), and the 111 Project of the Ministry of Education of China (B07049) for their support of this study. We appreciate the advice of Professor Peng Chen and Minghui Lu (Northwest A&F University). We thank Zelong Li (Northwest A&F University) for improving the phylogenetic tree, Dr Hua Zhao and Dr Fengping Yuan (State Key Laboratory of Crop Stress Biology) for their support in confocal microscope.

Author contributions

Z.S.K., X.J.W., and X.M.Z. conceived the framework of the paper; G.G. and P.F.G. contributed original ideas. Y.Q.Z., S.Y.G., M.L., F.Z., C.W., and H.K.L. performed the experiments; Y.Q.Z. analyzed the data. X.M.Z. and Y.Q.Z. wrote the article.

Data availability

The Ensembl Plants database was used to search the genome sequence data used in this paper, with accession numbers as follows: CaREM1.4 (PHT78353) and CaRIN4-12 (PHT66856).

Conflict of interest statement

The authors declare no conflicting of interest.

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