The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant Defenses
Abstract
:1. Introduction
2. Results
2.1. MaCFEM85 Was Most Closely Related to Non-Pathogenic Fungal CFEMs and Localized to the Plasma Membrane
2.2. MaCFEM85 Was Upregulated during Interactions with M. sativa
2.3. MaCFEM85 Interacts with MsWAK16 In Vitro and In Vivo
2.4. Key Sites for MaCFEM85 and MsWAK16 Interactions
2.5. The Interaction of MaCFEM85 with MsWAK16 Activates the Plant Immune Response
2.5.1. Evaluating the Role of MaCFEM85 and MsWAK16 in Disease Resistance against B. cinerea
2.5.2. Evaluating the Role of MaCFEM85 and MsWAK16 in Aphid Defense in N. benthamiana
2.5.3. Evaluating the Role of MaCFEM85 and MsWAK16 in Hormone Accumulation and Hormone-Related Gene Expression
3. Discussion
3.1. The Conserved CFEM Protein Motif Serves Multiple Functions in Fungal Species
3.2. Disulfide Bonds Are Important Structures for Protein Function
3.3. The Interaction of MaCFEM85 with MsWAK16 Activated Plant Defenses
4. Materials and Methods
4.1. Fungal Strains, Plant Materials, and Culturing Methods
4.2. qRT-PCR and Plasmid Construction
4.3. Transient Expression of Proteins in N. benthamiana
4.4. Phylogenetic Analysis
4.5. Yeast Two-Hybrid Assays
4.6. BiFC Assay
4.7. GST Pull-Down Assay
4.8. Identification of the Key Site in MaCFEM85
4.9. Plant Resistance Assays
4.10. Aphid Performance Assays
4.11. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Cai, N.; Nong, X.; Liu, R.; McNeill, M.R.; Wang, G.; Zhang, Z.; Tu, X. The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant Defenses. Int. J. Mol. Sci. 2023, 24, 4037. https://doi.org/10.3390/ijms24044037
Cai N, Nong X, Liu R, McNeill MR, Wang G, Zhang Z, Tu X. The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant Defenses. International Journal of Molecular Sciences. 2023; 24(4):4037. https://doi.org/10.3390/ijms24044037
Chicago/Turabian StyleCai, Ni, Xiangqun Nong, Rong Liu, Mark Richard McNeill, Guangjun Wang, Zehua Zhang, and Xiongbing Tu. 2023. "The Conserved Cysteine-Rich Secretory Protein MaCFEM85 Interacts with MsWAK16 to Activate Plant Defenses" International Journal of Molecular Sciences 24, no. 4: 4037. https://doi.org/10.3390/ijms24044037