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Visualization of localized pathogen-Induced pH modulation in almond tissues infected by Colletotrichum acutatum using confocal scanning laser microscopy.
Phytopathology 2008; 98(11):1171-8P

Abstract

Modulation of pH within the host during infection of almond by the anthracnose pathogen Colletotrichum acutatum was studied using confocal scanning laser microscopy and the dual emission fluorescence indicator SNARF-1. This highly sensitive method allowed visualization of the spatial distribution of localized pathogen-induced pH modulation within and in proximity to fungal infection structures in host tissue at the cellular level. Ratiometric measurement of fluorescence at two emission wavelengths and in situ calibration allowed the quantification of pH ranges. After incubation of leaf epidermal tissue with SNARF-1, distinct alkaline (pH 8 to > or =9), red-spectrum (650 nm wave length) fluorescent zones developed as partial or complete halos around many fungal appressoria and in infection vesicles at 24 to 36 h after inoculation. In samples taken after 48 to 72 h, colonizing hyphae in the biotrophic phase and subsequently in the necrotrophic phase were also emitting the red fluorescence that extended into the surrounding host tissue, as also verified by depth analyses. Host epidermal cells were intact and apparently alive during the fungal alkalization process, with no visible disruption of cell structure. Generally, the pH of epidermal cells in noninoculated samples or in areas away from the infection in inoculated samples was lower than pH 7 with green (i.e., 500 to 550 nm wave length) fluorescence detected. Using standard electrodes, a significant increase in pH and ammonia concentration in leaf and fruit tissue was also measured but only at advanced stages of disease. In contrast, hyphae of the pathogen Alternaria alternata were mostly acidic and no change in fluorescence was found inside invaded host cells. The sequence of events in the C. acutatum-almond interaction includes penetration, production of ammonia by C. acutatum, and subsequent pH modulation within almond epidermal tissue to an alkaline environment that leads to further colonization of the host.

Authors+Show Affiliations

Department of Mycology, Real Jardín Botánico CSIC, Madrid, Spain.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

18943405

Citation

Diéguez-Uribeondo, J, et al. "Visualization of Localized pathogen-Induced pH Modulation in Almond Tissues Infected By Colletotrichum Acutatum Using Confocal Scanning Laser Microscopy." Phytopathology, vol. 98, no. 11, 2008, pp. 1171-8.
Diéguez-Uribeondo J, Förster H, Adaskaveg JE. Visualization of localized pathogen-Induced pH modulation in almond tissues infected by Colletotrichum acutatum using confocal scanning laser microscopy. Phytopathology. 2008;98(11):1171-8.
Diéguez-Uribeondo, J., Förster, H., & Adaskaveg, J. E. (2008). Visualization of localized pathogen-Induced pH modulation in almond tissues infected by Colletotrichum acutatum using confocal scanning laser microscopy. Phytopathology, 98(11), pp. 1171-8. doi:10.1094/PHYTO-98-11-1171.
Diéguez-Uribeondo J, Förster H, Adaskaveg JE. Visualization of Localized pathogen-Induced pH Modulation in Almond Tissues Infected By Colletotrichum Acutatum Using Confocal Scanning Laser Microscopy. Phytopathology. 2008;98(11):1171-8. PubMed PMID: 18943405.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Visualization of localized pathogen-Induced pH modulation in almond tissues infected by Colletotrichum acutatum using confocal scanning laser microscopy. AU - Diéguez-Uribeondo,J, AU - Förster,H, AU - Adaskaveg,J E, PY - 2008/10/24/pubmed PY - 2009/3/24/medline PY - 2008/10/24/entrez SP - 1171 EP - 8 JF - Phytopathology JO - Phytopathology VL - 98 IS - 11 N2 - Modulation of pH within the host during infection of almond by the anthracnose pathogen Colletotrichum acutatum was studied using confocal scanning laser microscopy and the dual emission fluorescence indicator SNARF-1. This highly sensitive method allowed visualization of the spatial distribution of localized pathogen-induced pH modulation within and in proximity to fungal infection structures in host tissue at the cellular level. Ratiometric measurement of fluorescence at two emission wavelengths and in situ calibration allowed the quantification of pH ranges. After incubation of leaf epidermal tissue with SNARF-1, distinct alkaline (pH 8 to > or =9), red-spectrum (650 nm wave length) fluorescent zones developed as partial or complete halos around many fungal appressoria and in infection vesicles at 24 to 36 h after inoculation. In samples taken after 48 to 72 h, colonizing hyphae in the biotrophic phase and subsequently in the necrotrophic phase were also emitting the red fluorescence that extended into the surrounding host tissue, as also verified by depth analyses. Host epidermal cells were intact and apparently alive during the fungal alkalization process, with no visible disruption of cell structure. Generally, the pH of epidermal cells in noninoculated samples or in areas away from the infection in inoculated samples was lower than pH 7 with green (i.e., 500 to 550 nm wave length) fluorescence detected. Using standard electrodes, a significant increase in pH and ammonia concentration in leaf and fruit tissue was also measured but only at advanced stages of disease. In contrast, hyphae of the pathogen Alternaria alternata were mostly acidic and no change in fluorescence was found inside invaded host cells. The sequence of events in the C. acutatum-almond interaction includes penetration, production of ammonia by C. acutatum, and subsequent pH modulation within almond epidermal tissue to an alkaline environment that leads to further colonization of the host. SN - 0031-949X UR - https://www.unboundmedicine.com/medline/citation/18943405/Visualization_of_localized_pathogen_Induced_pH_modulation_in_almond_tissues_infected_by_Colletotrichum_acutatum_using_confocal_scanning_laser_microscopy_ L2 - http://apsjournals.apsnet.org/doi/full/10.1094/PHYTO-98-11-1171?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -