Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli
Authors
Gašić, Katarina
Obradović, Mina
Kuzmanović, Nemanja
Zlatković, Nevena
Ivanović, Milan
Ristić, Lela
Obradović, Aleksa

Article (Published version)
Metadata
Show full item recordAbstract
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 pl...aques, 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.
Keywords:
Acidovorax citrulli / bacteriophage / genome analysis / host specificity / phage therapy / biocontrolSource:
Frontiers in Microbiology, 2022, 12, 803789Publisher:
- Frontiers Media S.A.
Funding / projects:
- Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 200010 (Institute for Plant Protection and Environment, Belgrade) (RS-200010)
- Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 200116 (University of Belgrade, Faculty of Agriculture) (RS-200116)
- Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 200042 (University of Belgrade, Institute of Molecular Genetics and Genetic Engineering) (RS-200042)
- Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – Projektnummer 429677233
DOI: 10.3389/fmicb.2021.803789
ISSN: 1664-302X
PubMed: 35185829
WoS: 000759925600001
Scopus: 2-s2.0-85124755022
Collections
Institution/Community
IZBISTY - JOUR AU - Gašić, Katarina AU - Obradović, Mina AU - Kuzmanović, Nemanja AU - Zlatković, Nevena AU - Ivanović, Milan AU - Ristić, Lela AU - Obradović, Aleksa PY - 2022 UR - https://plantarum.izbis.bg.ac.rs/handle/123456789/725 AB - 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. PB - Frontiers Media S.A. T2 - Frontiers in Microbiology T1 - Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli IS - 803789 VL - 12 DO - 10.3389/fmicb.2021.803789 ER -
@article{ author = "Gašić, Katarina and Obradović, Mina and Kuzmanović, Nemanja and Zlatković, Nevena and Ivanović, Milan and Ristić, Lela and Obradović, Aleksa", year = "2022", abstract = "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.", publisher = "Frontiers Media S.A.", journal = "Frontiers in Microbiology", title = "Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli", number = "803789", volume = "12", doi = "10.3389/fmicb.2021.803789" }
Gašić, K., Obradović, M., Kuzmanović, N., Zlatković, N., Ivanović, M., Ristić, L.,& Obradović, A.. (2022). Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli. in Frontiers in Microbiology Frontiers Media S.A.., 12(803789). https://doi.org/10.3389/fmicb.2021.803789
Gašić K, Obradović M, Kuzmanović N, Zlatković N, Ivanović M, Ristić L, Obradović A. Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli. in Frontiers in Microbiology. 2022;12(803789). doi:10.3389/fmicb.2021.803789 .
Gašić, Katarina, Obradović, Mina, Kuzmanović, Nemanja, Zlatković, Nevena, Ivanović, Milan, Ristić, Lela, Obradović, Aleksa, "Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli" in Frontiers in Microbiology, 12, no. 803789 (2022), https://doi.org/10.3389/fmicb.2021.803789 . .