Cargando…

Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli

Bacterial fruit blotch and seedling blight, caused by Acidovorax citrulli, is one of the most destructive diseases of melon and watermelon in many countries. Pathogen-free seed and cultural practices are major pillars of the disease control. However, use of bacteriophages as natural biocontrol agent...

Descripción completa

Detalles Bibliográficos
Autores principales: Gašić, Katarina, Obradović, Mina, Kuzmanović, Nemanja, Zlatković, Nevena, Ivanović, Milan, Ristić, Danijela, Obradović, Aleksa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8851203/
https://www.ncbi.nlm.nih.gov/pubmed/35185829
http://dx.doi.org/10.3389/fmicb.2021.803789
_version_ 1784652776369815552
author Gašić, Katarina
Obradović, Mina
Kuzmanović, Nemanja
Zlatković, Nevena
Ivanović, Milan
Ristić, Danijela
Obradović, Aleksa
author_facet Gašić, Katarina
Obradović, Mina
Kuzmanović, Nemanja
Zlatković, Nevena
Ivanović, Milan
Ristić, Danijela
Obradović, Aleksa
author_sort Gašić, Katarina
collection PubMed
description Bacterial fruit blotch and seedling blight, caused by Acidovorax citrulli, is one of the most destructive diseases of melon and watermelon in many countries. Pathogen-free seed and cultural practices are major pillars of the disease control. However, use of bacteriophages as natural biocontrol agents might also contribute to the disease management. Therefore, we isolated 12 bacteriophages specific to A. citrulli, from phyllosphere and rhizosphere of diseased watermelon plants. The phage strains were characterized based on their host range, plaque and virion morphology, thermal inactivation point, adsorption rate, one step growth curve, restriction fragment length polymorphism (RFLP), and genomic analysis. Transmission electron microscopy of three phage strains indicated that they belong to the order Caudovirales, family Siphoviridae. All phages lysed 30 out of 32 tested A. citrulli strains isolated in Serbia, and did not lyse other less related bacterial species. They produced clear plaques, 2 mm in diameter, on bacterial lawns of different A. citrulli strains after 24 h of incubation. The thermal inactivation point was 66 or 67°C. They were stable at pH 5–9, but were sensitive to chloroform and inactivated in either 5 or 10 min exposure to ultraviolet (UV) light. RFLP analysis using EcoRI, BsmI and BamHI enzymes did not show genetic differences among the tested phages. Adsorption rate and one step growth curve were determined for the Acidovorax phage ACF1. Draft genome sequence of the ACF1 phage was 59.377 bp in size, with guanine-cytosine (GC) content 64.5%, including 89 open reading frames. This phage shared a very high genomic identity with Acidovorax phage ACPWH, isolated in South Korea. Evaluation of systemic nature of ACF1 strain showed that it can be absorbed by roots and translocated to upper parts of watermelon plants where it survived up to 10 days.
format Online
Article
Text
id pubmed-8851203
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-88512032022-02-18 Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli Gašić, Katarina Obradović, Mina Kuzmanović, Nemanja Zlatković, Nevena Ivanović, Milan Ristić, Danijela Obradović, Aleksa Front Microbiol Microbiology Bacterial fruit blotch and seedling blight, caused by Acidovorax citrulli, is one of the most destructive diseases of melon and watermelon in many countries. Pathogen-free seed and cultural practices are major pillars of the disease control. However, use of bacteriophages as natural biocontrol agents might also contribute to the disease management. Therefore, we isolated 12 bacteriophages specific to A. citrulli, from phyllosphere and rhizosphere of diseased watermelon plants. The phage strains were characterized based on their host range, plaque and virion morphology, thermal inactivation point, adsorption rate, one step growth curve, restriction fragment length polymorphism (RFLP), and genomic analysis. Transmission electron microscopy of three phage strains indicated that they belong to the order Caudovirales, family Siphoviridae. All phages lysed 30 out of 32 tested A. citrulli strains isolated in Serbia, and did not lyse other less related bacterial species. They produced clear plaques, 2 mm in diameter, on bacterial lawns of different A. citrulli strains after 24 h of incubation. The thermal inactivation point was 66 or 67°C. They were stable at pH 5–9, but were sensitive to chloroform and inactivated in either 5 or 10 min exposure to ultraviolet (UV) light. RFLP analysis using EcoRI, BsmI and BamHI enzymes did not show genetic differences among the tested phages. Adsorption rate and one step growth curve were determined for the Acidovorax phage ACF1. Draft genome sequence of the ACF1 phage was 59.377 bp in size, with guanine-cytosine (GC) content 64.5%, including 89 open reading frames. This phage shared a very high genomic identity with Acidovorax phage ACPWH, isolated in South Korea. Evaluation of systemic nature of ACF1 strain showed that it can be absorbed by roots and translocated to upper parts of watermelon plants where it survived up to 10 days. Frontiers Media S.A. 2022-02-03 /pmc/articles/PMC8851203/ /pubmed/35185829 http://dx.doi.org/10.3389/fmicb.2021.803789 Text en Copyright © 2022 Gašić, Obradović, Kuzmanović, Zlatković, Ivanović, Ristić and Obradović. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Gašić, Katarina
Obradović, Mina
Kuzmanović, Nemanja
Zlatković, Nevena
Ivanović, Milan
Ristić, Danijela
Obradović, Aleksa
Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli
title Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli
title_full Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli
title_fullStr Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli
title_full_unstemmed Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli
title_short Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli
title_sort isolation, characterization and draft genome analysis of bacteriophages infecting acidovorax citrulli
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8851203/
https://www.ncbi.nlm.nih.gov/pubmed/35185829
http://dx.doi.org/10.3389/fmicb.2021.803789
work_keys_str_mv AT gasickatarina isolationcharacterizationanddraftgenomeanalysisofbacteriophagesinfectingacidovoraxcitrulli
AT obradovicmina isolationcharacterizationanddraftgenomeanalysisofbacteriophagesinfectingacidovoraxcitrulli
AT kuzmanovicnemanja isolationcharacterizationanddraftgenomeanalysisofbacteriophagesinfectingacidovoraxcitrulli
AT zlatkovicnevena isolationcharacterizationanddraftgenomeanalysisofbacteriophagesinfectingacidovoraxcitrulli
AT ivanovicmilan isolationcharacterizationanddraftgenomeanalysisofbacteriophagesinfectingacidovoraxcitrulli
AT risticdanijela isolationcharacterizationanddraftgenomeanalysisofbacteriophagesinfectingacidovoraxcitrulli
AT obradovicaleksa isolationcharacterizationanddraftgenomeanalysisofbacteriophagesinfectingacidovoraxcitrulli