Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli
2022
Аутори
Gasić, KatarinaObradović, Mina
Kuzmanović, Nemanja
Zlatković, Nevena
Ivanović, Milan
Ristić, Danijela
Obradović, Aleksa
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
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 degrees 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.
Кључне речи:
phage therapy / host specificity / genome analysis / biocontrol / bacteriophage / Acidovorax citrulliИзвор:
Frontiers in Microbiology, 2022, 12Издавач:
- Frontiers Media Sa, Lausanne
Финансирање / пројекти:
- Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) [429677233]
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200010 (Институт за заштиту биља и животну средину, Београд) (RS-MESTD-inst-2020-200010)
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200116 (Универзитет у Београду, Пољопривредни факултет) (RS-MESTD-inst-2020-200116)
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200042 (Универзитет у Београду, Институт за молекуларну генетику и генетичко инжењерство) (RS-MESTD-inst-2020-200042)
DOI: 10.3389/fmicb.2021.803789
ISSN: 1664-302X
WoS: 000759925600001
Scopus: 2-s2.0-85124755022
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Gasić, Katarina AU - Obradović, Mina AU - Kuzmanović, Nemanja AU - Zlatković, Nevena AU - Ivanović, Milan AU - Ristić, Danijela AU - Obradović, Aleksa PY - 2022 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/1571 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 degrees 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 Sa, Lausanne T2 - Frontiers in Microbiology T1 - Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli VL - 12 DO - 10.3389/fmicb.2021.803789 ER -
@article{ author = "Gasić, Katarina and Obradović, Mina and Kuzmanović, Nemanja and Zlatković, Nevena and Ivanović, Milan and Ristić, Danijela 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 degrees 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 Sa, Lausanne", journal = "Frontiers in Microbiology", title = "Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli", volume = "12", doi = "10.3389/fmicb.2021.803789" }
Gasić, K., Obradović, M., Kuzmanović, N., Zlatković, N., Ivanović, M., Ristić, D.,& Obradović, A.. (2022). Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli. in Frontiers in Microbiology Frontiers Media Sa, Lausanne., 12. https://doi.org/10.3389/fmicb.2021.803789
Gasić K, Obradović M, Kuzmanović N, Zlatković N, Ivanović M, Ristić D, Obradović A. Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli. in Frontiers in Microbiology. 2022;12. doi:10.3389/fmicb.2021.803789 .
Gasić, Katarina, Obradović, Mina, Kuzmanović, Nemanja, Zlatković, Nevena, Ivanović, Milan, Ristić, Danijela, Obradović, Aleksa, "Isolation, Characterization and Draft Genome Analysis of Bacteriophages Infecting Acidovorax citrulli" in Frontiers in Microbiology, 12 (2022), https://doi.org/10.3389/fmicb.2021.803789 . .