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Functional and Pharmacological Analyses of the Role of Penicillium digitatum Proteases on Virulence

Penicillium digitatum is the major postharvest pathogen of citrus fruit under Mediterranean climate conditions. Previous results have shown that proteases is the largest enzyme family induced by P. digitatum during fruit infection. In the present work, we addressed the study of the role of P. digita...

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Autores principales: Ballester, Ana-Rosa, López-Pérez, Mario, de la Fuente, Beatriz, González-Candelas, Luis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6680461/
https://www.ncbi.nlm.nih.gov/pubmed/31336863
http://dx.doi.org/10.3390/microorganisms7070198
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author Ballester, Ana-Rosa
López-Pérez, Mario
de la Fuente, Beatriz
González-Candelas, Luis
author_facet Ballester, Ana-Rosa
López-Pérez, Mario
de la Fuente, Beatriz
González-Candelas, Luis
author_sort Ballester, Ana-Rosa
collection PubMed
description Penicillium digitatum is the major postharvest pathogen of citrus fruit under Mediterranean climate conditions. Previous results have shown that proteases is the largest enzyme family induced by P. digitatum during fruit infection. In the present work, we addressed the study of the role of P. digitatum’s proteases in virulence following two complementary approaches. In the first approach, we undertook the functional characterization of the P. digitatum prtT gene, which codes for a putative transcription factor previously shown to regulate extracellular proteases in other filamentous fungi. Deletion of prtT caused a significant loss in secreted protease activity during in vitro growth assays. However, there was no effect on virulence. Gene expression of the two major secreted acid proteases was barely affected in the ΔprtT deletant during infection of citrus fruit. Hence, no conclusion could be drawn on the role of these secreted acidic proteases on the virulence of P. digitatum. In the second approach, we studied the effect of different protease inhibitors and chelators on virulence. Co-inoculation of citrus fruit with P. digitatum conidia and a cocktail of protease inhibitors resulted in almost a complete absence of disease development. Analysis of individual inhibitors revealed that the metalloprotease inhibitor, 1,10-phenanthroline, was responsible for the observed effect. The application of metal ions reverted the protective effect caused by the metallopeptidase inhibitor. These results may set the basis for the development of new alternative treatments to combat this important postharvest pathogen.
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spelling pubmed-66804612019-08-09 Functional and Pharmacological Analyses of the Role of Penicillium digitatum Proteases on Virulence Ballester, Ana-Rosa López-Pérez, Mario de la Fuente, Beatriz González-Candelas, Luis Microorganisms Article Penicillium digitatum is the major postharvest pathogen of citrus fruit under Mediterranean climate conditions. Previous results have shown that proteases is the largest enzyme family induced by P. digitatum during fruit infection. In the present work, we addressed the study of the role of P. digitatum’s proteases in virulence following two complementary approaches. In the first approach, we undertook the functional characterization of the P. digitatum prtT gene, which codes for a putative transcription factor previously shown to regulate extracellular proteases in other filamentous fungi. Deletion of prtT caused a significant loss in secreted protease activity during in vitro growth assays. However, there was no effect on virulence. Gene expression of the two major secreted acid proteases was barely affected in the ΔprtT deletant during infection of citrus fruit. Hence, no conclusion could be drawn on the role of these secreted acidic proteases on the virulence of P. digitatum. In the second approach, we studied the effect of different protease inhibitors and chelators on virulence. Co-inoculation of citrus fruit with P. digitatum conidia and a cocktail of protease inhibitors resulted in almost a complete absence of disease development. Analysis of individual inhibitors revealed that the metalloprotease inhibitor, 1,10-phenanthroline, was responsible for the observed effect. The application of metal ions reverted the protective effect caused by the metallopeptidase inhibitor. These results may set the basis for the development of new alternative treatments to combat this important postharvest pathogen. MDPI 2019-07-12 /pmc/articles/PMC6680461/ /pubmed/31336863 http://dx.doi.org/10.3390/microorganisms7070198 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ballester, Ana-Rosa
López-Pérez, Mario
de la Fuente, Beatriz
González-Candelas, Luis
Functional and Pharmacological Analyses of the Role of Penicillium digitatum Proteases on Virulence
title Functional and Pharmacological Analyses of the Role of Penicillium digitatum Proteases on Virulence
title_full Functional and Pharmacological Analyses of the Role of Penicillium digitatum Proteases on Virulence
title_fullStr Functional and Pharmacological Analyses of the Role of Penicillium digitatum Proteases on Virulence
title_full_unstemmed Functional and Pharmacological Analyses of the Role of Penicillium digitatum Proteases on Virulence
title_short Functional and Pharmacological Analyses of the Role of Penicillium digitatum Proteases on Virulence
title_sort functional and pharmacological analyses of the role of penicillium digitatum proteases on virulence
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6680461/
https://www.ncbi.nlm.nih.gov/pubmed/31336863
http://dx.doi.org/10.3390/microorganisms7070198
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