Determination of Salmonella enterica Leaf Internalization Varies Substantially According to the Method and Conditions Used to Assess Bacterial Localization

In a previous study, comparing the internalization of S. enterica serovar Typhimurium in various leaves by confocal microscopy, we have demonstrated that the pathogen failed to internalize tomato leaves. Numerous reasons may account for these findings, yet one such factor might be the methodology em...

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Autores principales:	Madhvi Chahar, Yulia Kroupitski, Rachel Gollop, Eduard Belausov, Maeli Melotto, Shlomo Sela-Saldinger
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	tomato lettuce Arabidopsis internalization attachment colonization Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/4740eef7e14c46328b8c0efb20a82db2
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id	oai:doaj.org-article:4740eef7e14c46328b8c0efb20a82db2
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spelling	oai:doaj.org-article:4740eef7e14c46328b8c0efb20a82db22021-11-05T16:24:14ZDetermination of Salmonella enterica Leaf Internalization Varies Substantially According to the Method and Conditions Used to Assess Bacterial Localization1664-302X10.3389/fmicb.2021.622068https://doaj.org/article/4740eef7e14c46328b8c0efb20a82db22021-10-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fmicb.2021.622068/fullhttps://doaj.org/toc/1664-302XIn a previous study, comparing the internalization of S. enterica serovar Typhimurium in various leaves by confocal microscopy, we have demonstrated that the pathogen failed to internalize tomato leaves. Numerous reasons may account for these findings, yet one such factor might be the methodology employed to quantify leaf internalization. To this end, we have systematically studied leaf localization of a Green-fluorescent protein-labeled Salmonella strain in tomato, lettuce, and Arabidopsis leaves by surface sterilization and enumeration of the surviving bacteria, side by side, with confocal microscopy observations. Leaf sterilization was performed using either sodium hypochlorite, silver nitrate, or ethanol for 1 to 7min. The level of internalization varied according to the type of disinfectant used for surface sterilization and the treatment time. Treatment of tomato leaves with 70% ethanol for up to 7min suggested possible internalization of Salmonella, while confocal microscopy showed no internalization. In the case of in lettuce and Arabidopsis leaves, both the plate-count technique and confocal microscopy demonstrated considerable Salmonella internalization thought different sterilization conditions resulted in variations in the internalization levels. Our findings highlighted the dependency of the internalization results on the specific disinfection protocol used to determine bacterial localization. The results underscore the importance of confocal microscopy in validating a particular surface sterilization protocol whenever a new pair of bacterial strain and plant cultivar is studied.Madhvi ChaharYulia KroupitskiRachel GollopEduard BelausovMaeli MelottoShlomo Sela-SaldingerFrontiers Media S.A.articletomatolettuceArabidopsisinternalizationattachmentcolonizationMicrobiologyQR1-502ENFrontiers in Microbiology, Vol 12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	tomato lettuce Arabidopsis internalization attachment colonization Microbiology QR1-502
spellingShingle	tomato lettuce Arabidopsis internalization attachment colonization Microbiology QR1-502 Madhvi Chahar Yulia Kroupitski Rachel Gollop Eduard Belausov Maeli Melotto Shlomo Sela-Saldinger Determination of Salmonella enterica Leaf Internalization Varies Substantially According to the Method and Conditions Used to Assess Bacterial Localization
description	In a previous study, comparing the internalization of S. enterica serovar Typhimurium in various leaves by confocal microscopy, we have demonstrated that the pathogen failed to internalize tomato leaves. Numerous reasons may account for these findings, yet one such factor might be the methodology employed to quantify leaf internalization. To this end, we have systematically studied leaf localization of a Green-fluorescent protein-labeled Salmonella strain in tomato, lettuce, and Arabidopsis leaves by surface sterilization and enumeration of the surviving bacteria, side by side, with confocal microscopy observations. Leaf sterilization was performed using either sodium hypochlorite, silver nitrate, or ethanol for 1 to 7min. The level of internalization varied according to the type of disinfectant used for surface sterilization and the treatment time. Treatment of tomato leaves with 70% ethanol for up to 7min suggested possible internalization of Salmonella, while confocal microscopy showed no internalization. In the case of in lettuce and Arabidopsis leaves, both the plate-count technique and confocal microscopy demonstrated considerable Salmonella internalization thought different sterilization conditions resulted in variations in the internalization levels. Our findings highlighted the dependency of the internalization results on the specific disinfection protocol used to determine bacterial localization. The results underscore the importance of confocal microscopy in validating a particular surface sterilization protocol whenever a new pair of bacterial strain and plant cultivar is studied.
format	article
author	Madhvi Chahar Yulia Kroupitski Rachel Gollop Eduard Belausov Maeli Melotto Shlomo Sela-Saldinger
author_facet	Madhvi Chahar Yulia Kroupitski Rachel Gollop Eduard Belausov Maeli Melotto Shlomo Sela-Saldinger
author_sort	Madhvi Chahar
title	Determination of Salmonella enterica Leaf Internalization Varies Substantially According to the Method and Conditions Used to Assess Bacterial Localization
title_short	Determination of Salmonella enterica Leaf Internalization Varies Substantially According to the Method and Conditions Used to Assess Bacterial Localization
title_full	Determination of Salmonella enterica Leaf Internalization Varies Substantially According to the Method and Conditions Used to Assess Bacterial Localization
title_fullStr	Determination of Salmonella enterica Leaf Internalization Varies Substantially According to the Method and Conditions Used to Assess Bacterial Localization
title_full_unstemmed	Determination of Salmonella enterica Leaf Internalization Varies Substantially According to the Method and Conditions Used to Assess Bacterial Localization
title_sort	determination of salmonella enterica leaf internalization varies substantially according to the method and conditions used to assess bacterial localization
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/4740eef7e14c46328b8c0efb20a82db2
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Determination of Salmonella enterica Leaf Internalization Varies Substantially According to the Method and Conditions Used to Assess Bacterial Localization

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