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Interaction of antifungal peptide BP15 with Stemphylium vesicarium, the causal agent of brown spot of pear

Peptide BP15 has shown antifungal activity against several plant pathogenic fungi, including Stemphylium vesicarium, the causal agent of brown spot of pear. BP15 inhibits the germination, growth and sporulation of S. vesicarium and displays post-infection activity by stopping fungal infection in pear leaves. In this work, live-cell imaging was undertaken to understand the antifungal mechanism of BP15. A double-staining method based on the combination of calcofluor white and SYTOX green coupled with epifluorescence microscopy was used to investigate fungal cell permeabilization and alterations in fungal growth induced by BP15. GFP-transformants of S. vesicarium were obtained and exposed to rhodamine-labelled BP15. Confocal laser microscopy provided evidence of peptide internalization by hyphae, resulting in fungal cell disorganization and death. S. vesicarium membrane permeabilization by BP15 was found to be peptide-concentration dependent. BP15 at MIC and sub-MIC concentrations (10 and 5 μM, respectively) inhibited S. vesicarium growth and produced morphological alterations to germ tubes, with slow and discontinuous compromise of fungal cell membranes. Fungal cell membrane disruption was immediately induced by BP15 at 100 μM, and this was accompanied by rapid peptide internalization by S. vesicarium hyphae. Peptide BP15 interacted with germ tubes and hyphae of S. vesicarium but not with conidial cells

This research was supported in part by grants from the Ministerio de Educaci on y Ciencia (AGL2009-09829/AGR and AGL2011-30354-C02-01) of Spain

© Fungal Biology, 2016, vol. 120, núm. 1, p. 61-71

Elsevier

Author: Puig Garcia, Mireia
Moragrega i Garcia, Concepció
Ruz Estévez, Lídia
Calderón, Claudia E.
Cazorla, Francisco M.
Montesinos Seguí, Emilio
Llorente i Cabratosa, Isidre
Date: 2016
Abstract: Peptide BP15 has shown antifungal activity against several plant pathogenic fungi, including Stemphylium vesicarium, the causal agent of brown spot of pear. BP15 inhibits the germination, growth and sporulation of S. vesicarium and displays post-infection activity by stopping fungal infection in pear leaves. In this work, live-cell imaging was undertaken to understand the antifungal mechanism of BP15. A double-staining method based on the combination of calcofluor white and SYTOX green coupled with epifluorescence microscopy was used to investigate fungal cell permeabilization and alterations in fungal growth induced by BP15. GFP-transformants of S. vesicarium were obtained and exposed to rhodamine-labelled BP15. Confocal laser microscopy provided evidence of peptide internalization by hyphae, resulting in fungal cell disorganization and death. S. vesicarium membrane permeabilization by BP15 was found to be peptide-concentration dependent. BP15 at MIC and sub-MIC concentrations (10 and 5 μM, respectively) inhibited S. vesicarium growth and produced morphological alterations to germ tubes, with slow and discontinuous compromise of fungal cell membranes. Fungal cell membrane disruption was immediately induced by BP15 at 100 μM, and this was accompanied by rapid peptide internalization by S. vesicarium hyphae. Peptide BP15 interacted with germ tubes and hyphae of S. vesicarium but not with conidial cells
This research was supported in part by grants from the Ministerio de Educaci on y Ciencia (AGL2009-09829/AGR and AGL2011-30354-C02-01) of Spain
Format: application/pdf
ISSN: 1878-6146
Document access: http://hdl.handle.net/10256/12345
Language: eng
Publisher: Elsevier
Collection: MICINN/PN 2010-2013/AGL2009-09829
Reproducció digital del document publicat a: http://dx.doi.org/10.1016/j.funbio.2015.10.007
Articles publicats (D-EQATA)
Is part of: © Fungal Biology, 2016, vol. 120, núm. 1, p. 61-71
Rights: Tots els drets reservats
Subject: Fungicides
Perera -- Malalties i plagues
Pear -- Diseases and pests
Fongs en l’agricultura
Fungi in agriculture
Title: Interaction of antifungal peptide BP15 with Stemphylium vesicarium, the causal agent of brown spot of pear
Type: info:eu-repo/semantics/article
Repository: DUGiDocs

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