Erwinia amylovora CRISPR elements provide new tools for evaluating strain diversity and for microbial source tracking.

Clustered regularly interspaced short palindromic repeats (CRISPRs) comprise a family of short DNA repeat sequences that are separated by non repetitive spacer sequences and, in combination with a suite of Cas proteins, are thought to function as an adaptive immune system against invading DNA. The n...

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Main Authors: Gayle C McGhee, George W Sundin
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2012-01-01
Series:PLoS ONE
Online Access:http://europepmc.org/articles/PMC3409226?pdf=render
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author Gayle C McGhee
George W Sundin
author_facet Gayle C McGhee
George W Sundin
author_sort Gayle C McGhee
collection DOAJ
description Clustered regularly interspaced short palindromic repeats (CRISPRs) comprise a family of short DNA repeat sequences that are separated by non repetitive spacer sequences and, in combination with a suite of Cas proteins, are thought to function as an adaptive immune system against invading DNA. The number of CRISPR arrays in a bacterial chromosome is variable, and the content of each array can differ in both repeat number and in the presence or absence of specific spacers. We utilized a comparative sequence analysis of CRISPR arrays of the plant pathogen Erwinia amylovora to uncover previously unknown genetic diversity in this species. A total of 85 E. amylovora strains varying in geographic isolation (North America, Europe, New Zealand, and the Middle East), host range, plasmid content, and streptomycin sensitivity/resistance were evaluated for CRISPR array number and spacer variability. From these strains, 588 unique spacers were identified in the three CRISPR arrays present in E. amylovora, and these arrays could be categorized into 20, 17, and 2 patterns types, respectively. Analysis of the relatedness of spacer content differentiated most apple and pear strains isolated in the eastern U.S. from western U.S. strains. In addition, we identified North American strains that shared CRISPR genotypes with strains isolated on other continents. E. amylovora strains from Rubus and Indian hawthorn contained mostly unique spacers compared to apple and pear strains, while strains from loquat shared 79% of spacers with apple and pear strains. Approximately 23% of the spacers matched known sequences, with 16% targeting plasmids and 5% targeting bacteriophage. The plasmid pEU30, isolated in E. amylovora strains from the western U.S., was targeted by 55 spacers. Lastly, we used spacer patterns and content to determine that streptomycin-resistant strains of E. amylovora from Michigan were low in diversity and matched corresponding streptomycin-sensitive strains from the background population.
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spelling doaj.art-a65c0eacfeb94a33b40b24ccbe7767432022-12-21T17:25:07ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0177e4170610.1371/journal.pone.0041706Erwinia amylovora CRISPR elements provide new tools for evaluating strain diversity and for microbial source tracking.Gayle C McGheeGeorge W SundinClustered regularly interspaced short palindromic repeats (CRISPRs) comprise a family of short DNA repeat sequences that are separated by non repetitive spacer sequences and, in combination with a suite of Cas proteins, are thought to function as an adaptive immune system against invading DNA. The number of CRISPR arrays in a bacterial chromosome is variable, and the content of each array can differ in both repeat number and in the presence or absence of specific spacers. We utilized a comparative sequence analysis of CRISPR arrays of the plant pathogen Erwinia amylovora to uncover previously unknown genetic diversity in this species. A total of 85 E. amylovora strains varying in geographic isolation (North America, Europe, New Zealand, and the Middle East), host range, plasmid content, and streptomycin sensitivity/resistance were evaluated for CRISPR array number and spacer variability. From these strains, 588 unique spacers were identified in the three CRISPR arrays present in E. amylovora, and these arrays could be categorized into 20, 17, and 2 patterns types, respectively. Analysis of the relatedness of spacer content differentiated most apple and pear strains isolated in the eastern U.S. from western U.S. strains. In addition, we identified North American strains that shared CRISPR genotypes with strains isolated on other continents. E. amylovora strains from Rubus and Indian hawthorn contained mostly unique spacers compared to apple and pear strains, while strains from loquat shared 79% of spacers with apple and pear strains. Approximately 23% of the spacers matched known sequences, with 16% targeting plasmids and 5% targeting bacteriophage. The plasmid pEU30, isolated in E. amylovora strains from the western U.S., was targeted by 55 spacers. Lastly, we used spacer patterns and content to determine that streptomycin-resistant strains of E. amylovora from Michigan were low in diversity and matched corresponding streptomycin-sensitive strains from the background population.http://europepmc.org/articles/PMC3409226?pdf=render
spellingShingle Gayle C McGhee
George W Sundin
Erwinia amylovora CRISPR elements provide new tools for evaluating strain diversity and for microbial source tracking.
PLoS ONE
title Erwinia amylovora CRISPR elements provide new tools for evaluating strain diversity and for microbial source tracking.
title_full Erwinia amylovora CRISPR elements provide new tools for evaluating strain diversity and for microbial source tracking.
title_fullStr Erwinia amylovora CRISPR elements provide new tools for evaluating strain diversity and for microbial source tracking.
title_full_unstemmed Erwinia amylovora CRISPR elements provide new tools for evaluating strain diversity and for microbial source tracking.
title_short Erwinia amylovora CRISPR elements provide new tools for evaluating strain diversity and for microbial source tracking.
title_sort erwinia amylovora crispr elements provide new tools for evaluating strain diversity and for microbial source tracking
url http://europepmc.org/articles/PMC3409226?pdf=render
work_keys_str_mv AT gaylecmcghee erwiniaamylovoracrisprelementsprovidenewtoolsforevaluatingstraindiversityandformicrobialsourcetracking
AT georgewsundin erwiniaamylovoracrisprelementsprovidenewtoolsforevaluatingstraindiversityandformicrobialsourcetracking