Suppression of Chitin-Triggered Immunity by a New Fungal Chitin-Binding Effector Resulting from Alternative Splicing of a Chitin Deacetylase Gene
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plants, Fungi, Bacteria, and Culture Conditions
2.2. Sequence Analysis and Protein Modelling
2.3. RNA Extraction and cDNA Synthesis
2.4. Plasmid Construction
2.5. Protein Expression and Purification
2.6. Chitin-Binding Activity Assays
2.7. Chitinase Activity and Chitin-Triggered Oxidative Burst Assay
2.8. Transformation by Growth onto Agroinfiltrated Tissues (TGAT) and Confocal Laser Scanning Microscopy (CLSM)
2.9. Real Time-Quantitative Polymerase Chain Reaction (RT-qPCR)
3. Results
3.1. Alternative Splicing of the PxCDA Gene Results in a Chitin-Binding Protein
3.2. PxCDA3 Protein Prevents Chitin Recognition by Sequestering Immunogenic Oligomers
3.3. PxCHBE Protein Is Deployed at the Plant Papilla Where Chitin Is Densely Accumulated
3.4. CHBE Is a Novel Fungal Effector That May Be Present in Other Powdery Mildew Fungi
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Assay | Substrate a | Products (µg mL−1) b | ||||||
---|---|---|---|---|---|---|---|---|
Mono | Di | Tri | Tetra | Penta | Hexa | Hepta | ||
Experiment 1 | (GlcNAc)1–7 | - c | - | - | - | 57 | 235 | Nd d |
Experiment 2 | (GlcNAc)1–7 | - | - | - | - | 50 | 186 | nd |
Experiment 3 | (GlcNAc)1–7 | - | - | - | - | 49 | 193 | nd |
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Martínez-Cruz, J.M.; Polonio, Á.; Ruiz-Jiménez, L.; Vielba-Fernández, A.; Hierrezuelo, J.; Romero, D.; de Vicente, A.; Fernández-Ortuño, D.; Pérez-García, A. Suppression of Chitin-Triggered Immunity by a New Fungal Chitin-Binding Effector Resulting from Alternative Splicing of a Chitin Deacetylase Gene. J. Fungi 2022, 8, 1022. https://doi.org/10.3390/jof8101022
Martínez-Cruz JM, Polonio Á, Ruiz-Jiménez L, Vielba-Fernández A, Hierrezuelo J, Romero D, de Vicente A, Fernández-Ortuño D, Pérez-García A. Suppression of Chitin-Triggered Immunity by a New Fungal Chitin-Binding Effector Resulting from Alternative Splicing of a Chitin Deacetylase Gene. Journal of Fungi. 2022; 8(10):1022. https://doi.org/10.3390/jof8101022
Chicago/Turabian StyleMartínez-Cruz, Jesús M., Álvaro Polonio, Laura Ruiz-Jiménez, Alejandra Vielba-Fernández, Jesús Hierrezuelo, Diego Romero, Antonio de Vicente, Dolores Fernández-Ortuño, and Alejandro Pérez-García. 2022. "Suppression of Chitin-Triggered Immunity by a New Fungal Chitin-Binding Effector Resulting from Alternative Splicing of a Chitin Deacetylase Gene" Journal of Fungi 8, no. 10: 1022. https://doi.org/10.3390/jof8101022