In Situ Identification of Plant-Invasive Bacteria with MALDI-TOF Mass Spectrometry

Rhizobia form a disparate collection of soil bacteria capable of reducing atmospheric nitrogen in symbiosis with legumes. The study of rhizobial populations in nature involves the collection of large numbers of nodules found on roots or stems of legumes, and the subsequent typing of nodule bacteria....

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Main Authors:	Ziegler, Dominik, Mariotti, Anna, Pflüger, Valentin, Saad, Maged, Vogel, Guido, Tonolla, Mauro, Perret, Xavier
Format:	Online
Language:	English
Published:	Public Library of Science 2012
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3355115/

id	pubmed-3355115
recordtype	oai_dc
spelling	pubmed-33551152012-05-21 In Situ Identification of Plant-Invasive Bacteria with MALDI-TOF Mass Spectrometry Ziegler, Dominik Mariotti, Anna Pflüger, Valentin Saad, Maged Vogel, Guido Tonolla, Mauro Perret, Xavier Research Article Rhizobia form a disparate collection of soil bacteria capable of reducing atmospheric nitrogen in symbiosis with legumes. The study of rhizobial populations in nature involves the collection of large numbers of nodules found on roots or stems of legumes, and the subsequent typing of nodule bacteria. To avoid the time-consuming steps of isolating and cultivating nodule bacteria prior to genotyping, a protocol of strain identification based on the comparison of MALDI-TOF MS spectra was established. In this procedure, plant nodules were considered as natural bioreactors that amplify clonal populations of nitrogen-fixing bacteroids. Following a simple isolation procedure, bacteroids were fingerprinted by analysing biomarker cellular proteins of 3 to 13 kDa using Matrix Assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) mass spectrometry. In total, bacteroids of more than 1,200 nodules collected from roots of three legumes of the Phaseoleae tribe (cowpea, soybean or siratro) were examined. Plants were inoculated with pure cultures of a slow-growing Bradyrhizobium japonicum strain G49, or either of two closely related and fast-growing Sinorhizobium fredii strains NGR234 and USDA257, or with mixed inoculants. In the fully automatic mode, correct identification of bacteroids was obtained for >97% of the nodules, and reached 100% with a minimal manual input in processing of spectra. These results showed that MALDI-TOF MS is a powerful tool for the identification of intracellular bacteria taken directly from plant tissues. Public Library of Science 2012-05-17 /pmc/articles/PMC3355115/ /pubmed/22615938 http://dx.doi.org/10.1371/journal.pone.0037189 Text en Ziegler et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Ziegler, Dominik Mariotti, Anna Pflüger, Valentin Saad, Maged Vogel, Guido Tonolla, Mauro Perret, Xavier
spellingShingle	Ziegler, Dominik Mariotti, Anna Pflüger, Valentin Saad, Maged Vogel, Guido Tonolla, Mauro Perret, Xavier In Situ Identification of Plant-Invasive Bacteria with MALDI-TOF Mass Spectrometry
author_facet	Ziegler, Dominik Mariotti, Anna Pflüger, Valentin Saad, Maged Vogel, Guido Tonolla, Mauro Perret, Xavier
author_sort	Ziegler, Dominik
title	In Situ Identification of Plant-Invasive Bacteria with MALDI-TOF Mass Spectrometry
title_short	In Situ Identification of Plant-Invasive Bacteria with MALDI-TOF Mass Spectrometry
title_full	In Situ Identification of Plant-Invasive Bacteria with MALDI-TOF Mass Spectrometry
title_fullStr	In Situ Identification of Plant-Invasive Bacteria with MALDI-TOF Mass Spectrometry
title_full_unstemmed	In Situ Identification of Plant-Invasive Bacteria with MALDI-TOF Mass Spectrometry
title_sort	in situ identification of plant-invasive bacteria with maldi-tof mass spectrometry
description	Rhizobia form a disparate collection of soil bacteria capable of reducing atmospheric nitrogen in symbiosis with legumes. The study of rhizobial populations in nature involves the collection of large numbers of nodules found on roots or stems of legumes, and the subsequent typing of nodule bacteria. To avoid the time-consuming steps of isolating and cultivating nodule bacteria prior to genotyping, a protocol of strain identification based on the comparison of MALDI-TOF MS spectra was established. In this procedure, plant nodules were considered as natural bioreactors that amplify clonal populations of nitrogen-fixing bacteroids. Following a simple isolation procedure, bacteroids were fingerprinted by analysing biomarker cellular proteins of 3 to 13 kDa using Matrix Assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) mass spectrometry. In total, bacteroids of more than 1,200 nodules collected from roots of three legumes of the Phaseoleae tribe (cowpea, soybean or siratro) were examined. Plants were inoculated with pure cultures of a slow-growing Bradyrhizobium japonicum strain G49, or either of two closely related and fast-growing Sinorhizobium fredii strains NGR234 and USDA257, or with mixed inoculants. In the fully automatic mode, correct identification of bacteroids was obtained for >97% of the nodules, and reached 100% with a minimal manual input in processing of spectra. These results showed that MALDI-TOF MS is a powerful tool for the identification of intracellular bacteria taken directly from plant tissues.
publisher	Public Library of Science
publishDate	2012
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3355115/
_version_	1611530790950141952

In Situ Identification of Plant-Invasive Bacteria with MALDI-TOF Mass Spectrometry

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