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...
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Frontiers Media S.A.
2022-02-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2021.803789/full |
| _version_ | 1818509821132931072 |
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| author | Katarina Gašić Mina Obradović Nemanja Kuzmanović Nevena Zlatković Milan Ivanović Danijela Ristić Aleksa Obradović |
| author_facet | Katarina Gašić Mina Obradović Nemanja Kuzmanović Nevena Zlatković Milan Ivanović Danijela Ristić Aleksa Obradović |
| author_sort | Katarina Gašić |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-10T22:50:41Z |
| format | Article |
| id | doaj.art-a1fbb24f896a4b18a6b5d6e3c406013e |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-10T22:50:41Z |
| publishDate | 2022-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-a1fbb24f896a4b18a6b5d6e3c406013e2022-12-22T01:30:26ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2022-02-011210.3389/fmicb.2021.803789803789Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulliKatarina Gašić0Mina Obradović1Nemanja Kuzmanović2Nevena Zlatković3Milan Ivanović4Danijela Ristić5Aleksa Obradović6Institute for Plant Protection and Environment, Belgrade, SerbiaInstitute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, SerbiaFederal Research Centre for Cultivated Plants, Institute for Plant Protection in Horticulture and Forests, Julius Kühn-Institut, Braunschweig, GermanyInstitute for Plant Protection and Environment, Belgrade, SerbiaFaculty of Agriculture, University of Belgrade, Belgrade, SerbiaInstitute for Plant Protection and Environment, Belgrade, SerbiaFaculty of Agriculture, University of Belgrade, Belgrade, SerbiaBacterial 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.https://www.frontiersin.org/articles/10.3389/fmicb.2021.803789/fullAcidovorax citrullibacteriophagegenome analysishost specificityphage therapybiocontrol |
| spellingShingle | Katarina Gašić Mina Obradović Nemanja Kuzmanović Nevena Zlatković Milan Ivanović Danijela Ristić Aleksa Obradović Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli Frontiers in Microbiology Acidovorax citrulli bacteriophage genome analysis host specificity phage therapy biocontrol |
| 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 | Acidovorax citrulli bacteriophage genome analysis host specificity phage therapy biocontrol |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2021.803789/full |
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