The RsRlpA Effector Is a Protease Inhibitor Promoting Rhizoctonia solani Virulence through Suppression of the Hypersensitive Response
Abstract
:1. Introduction
2. Results
2.1. The RsRlpA Contains a DPBB Fold and Is Highly Induced upon Early Infection
2.2. Heterologous Expression of the RsRlpA Promotes Fungal Infection
2.3. The RsRplA is Localized to the Cell Periphery and Suppresses Hypersensitive Response Induced by the Avr4/Cf4 Complex
2.4. The RsRlpA is an Active Protease Inhibitor Suppressing Oxidative Burst
2.5. The RsRplA Associates with a Plant Cathepsin
2.6. The Ser120 is Involved in RsRplA Ability to Suppress Cell Death
3. Discussion
4. Materials and Methods
4.1. Fungal Isolates and Growth Conditions
4.2. RNA Isolation and qRT-PCR Analysis
4.3. Construction of Cercospora Beticola Transgenic Strains and Infection Assays
4.4. Transgenic Arabidopsis Plants and Pathogenicity Assays
4.5. Effector Subcellular Localization and Hypersensitive Response Assays
4.6. Prediction of Protein 3D Structure, Production and Purification
4.7. Protease Inhibition and ROS Burst Suppression Assays
4.8. Pull-Down and Mass Spectrometry
4.9. Protein-Protein Interaction
4.10. Statistical Analysis
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
AG | Anastomosis group |
DAMP | Damage-associated molecular patterns |
CAZy | Carbohydrate-Active enZYmes |
DPBB | Double psi beta barrel |
DPI | Days post inoculation |
ETI | Effector-triggered immunity |
HR | Hypersensitive response |
PAMPs | Pathogen-associated molecular patterns |
PLCP | Papain-like cysteine protease |
PRR | Pattern recognition receptors |
PTI | Pattern-triggered immunity |
RlpA | Rare lipoprotein A |
ROS | Reactive oxygen species |
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Charova, S.N.; Dölfors, F.; Holmquist, L.; Moschou, P.N.; Dixelius, C.; Tzelepis, G. The RsRlpA Effector Is a Protease Inhibitor Promoting Rhizoctonia solani Virulence through Suppression of the Hypersensitive Response. Int. J. Mol. Sci. 2020, 21, 8070. https://doi.org/10.3390/ijms21218070
Charova SN, Dölfors F, Holmquist L, Moschou PN, Dixelius C, Tzelepis G. The RsRlpA Effector Is a Protease Inhibitor Promoting Rhizoctonia solani Virulence through Suppression of the Hypersensitive Response. International Journal of Molecular Sciences. 2020; 21(21):8070. https://doi.org/10.3390/ijms21218070
Chicago/Turabian StyleCharova, Spyridoula N., Fredrik Dölfors, Louise Holmquist, Panagiotis N. Moschou, Christina Dixelius, and Georgios Tzelepis. 2020. "The RsRlpA Effector Is a Protease Inhibitor Promoting Rhizoctonia solani Virulence through Suppression of the Hypersensitive Response" International Journal of Molecular Sciences 21, no. 21: 8070. https://doi.org/10.3390/ijms21218070