Abstract
Pseudocercospora fijiensis causes black Sigatoka disease, the most important threat to banana. The cell wall is crucial for fungal biological processes, including pathogenesis. Here, we performed cell wall proteomics analyses of two P. fijiensis strains, the highly virulent Oz2b, and the less virulent C1233 strains. Strains were starved from nitrogen to mimic the host environment. Interestingly, in vitro cultures of the C1233 strain grew faster than Oz2b in PDB medium, suggesting that C1233 survives outside the host better than the highly virulent Oz2b strain. Both strains were submitted to nitrogen starvation and the cell wall proteins were isolated and subjected to nano-HPLC–MS/MS. A total of 2686 proteins were obtained from which only 240 had a known function and thus, bioinformatics analyses were performed on this group. We found that 90 cell wall proteins were shared by both strains, 21 were unique for Oz2b and 39 for C1233. Shared proteins comprised 24 pathogenicity factors, including Avr4 and Ecp6, two effectors from P. fijiensis, while the unique proteins comprised 16 virulence factors in C1233 and 11 in Oz2b. The P. fijiensis cell wall proteome comprised canonical proteins, but thirty percent were atypical, a feature which in other phytopathogens has been interpreted as contamination. However, a comparison with the identities of atypical proteins in other reports suggests that the P. fijiensis proteins we detected were not contaminants. This is the first proteomics analysis of the P. fijiensis cell wall and our results expands the understanding of the fundamental biology of fungal phytopathogens and will help to decipher the molecular mechanisms of pathogenesis and virulence in P. fijiensis.
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Acknowledgements
The authors would like to thank Dr. Marco A. Villanueva, for his critical reading, as well as to the three anonymous reviewers, whose comments have helped to improve the quality of the data analysis in the manuscript. We would also wish to acknowledge E. Góngora-Castillo and A. Enríquez-Valencia for their advice regarding bioinformatics analysis. Y. Burgos-Canul was supported by scholarship No. 255427 for Ph. D. studies from CONACYT México. This work was funded by Grants 220957 from CONACyT and 247355 from FOMIX.
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YB-C, II-F and BC-C conceived, designed and wrote the paper; VL-V and APB gave support in the design of proteomic experiments; YB-C, performed cell wall isolation and protein extractions; MB and GM performed MS analysis, searched for protein IDs in the Uniprot database and edited the preliminary list of proteins; LB-A, MC-P and GM-P, provided P. fijiensis strains and technically supported the research; MT-S assisted with infection experiments, photography and sample preparation, II-F and BC-C; supervised all the work. All authors participated in the data analysis, wrote and read the manuscript.
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Burgos-Canul, Y.Y., Canto-Canché, B., Berezovski, M.V. et al. The cell wall proteome from two strains of Pseudocercospora fijiensis with differences in virulence. World J Microbiol Biotechnol 35, 105 (2019). https://doi.org/10.1007/s11274-019-2681-2
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DOI: https://doi.org/10.1007/s11274-019-2681-2