TY  - JOUR
AU  - Gašić, Katarina
AU  - Obradović, Mina
AU  - Kuzmanović, Nemanja
AU  - Zlatković, Nevena
AU  - Ivanović, Milan
AU  - Ristić, Danijela
AU  - Obradović, Aleksa
PY  - 2022
UR  - http://aspace.agrif.bg.ac.rs/handle/123456789/6013
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
SP  - 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, 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°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",
pages = "803789",
volume = "12",
doi = "10.3389/fmicb.2021.803789"
}

Gašić, 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 S.A.., 12, 803789.
https://doi.org/10.3389/fmicb.2021.803789

Gašić 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;12:803789.
doi:10.3389/fmicb.2021.803789 .

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" in Frontiers in Microbiology, 12 (2022):803789,
https://doi.org/10.3389/fmicb.2021.803789 . .

TY  - JOUR
AU  - Mirković, Nemanja
AU  - Obradović, Mina
AU  - O’Connor, Paula M.
AU  - Filipić, Brankica
AU  - Jovčić, Branko
AU  - Cotter, Paul D.
AU  - Kojić, Milan
PY  - 2021
UR  - http://aspace.agrif.bg.ac.rs/handle/123456789/5908
AB  - Screening for producers of potent antimicrobial peptides, resulted in the isolation of Bacillus cereus BGNM1 with strong antimicrobial activity against Listeria monocytogenes. Genome sequence analysis revealed that BGNM1 contains the gene cluster associated with the production of the lantibiotic, thusin, previously identified in B. thuringiensis. Purification of the antimicrobial activity confirmed that strain BGMN1 produces thusin. Both thusin sensitive and resistant strains were detected among clinical isolates of Streptococcus agalactiae. Random mutagenesis of a thusin sensitive strain, S. agalactiae B782, was performed in an attempt to identify the receptor protein for thusin. Three independent thusin resistant mutants were selected and their complete genomes sequenced. Comparative sequence analysis of these mutants with the WT strain revealed that duplication of a region encoding a 79 amino acids repeat in a C-protein α-antigen was a common difference, suggesting it to be responsible for increased resistance to thusin. Since induced thusin resistant mutants showed higher level of resistance than the naturally resistant B761 strain, complete genome sequencing of strain B761 was performed to check the integrity of the C-protein α-antigen-encoding gene. This analysis revealed that this gene is deleted in B761, providing further evidence that this protein promotes interaction of the thusin with receptor.
PB  - Springer Science and Business Media Deutschland GmbH
T2  - Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology
T1  - C-protein α-antigen modulates the lantibiotic thusin resistance in Streptococcus agalactiae
EP  - 1607
IS  - 10
SP  - 1595
VL  - 114
DO  - 10.1007/s10482-021-01626-3
ER  - 

@article{
author = "Mirković, Nemanja and Obradović, Mina and O’Connor, Paula M. and Filipić, Brankica and Jovčić, Branko and Cotter, Paul D. and Kojić, Milan",
year = "2021",
abstract = "Screening for producers of potent antimicrobial peptides, resulted in the isolation of Bacillus cereus BGNM1 with strong antimicrobial activity against Listeria monocytogenes. Genome sequence analysis revealed that BGNM1 contains the gene cluster associated with the production of the lantibiotic, thusin, previously identified in B. thuringiensis. Purification of the antimicrobial activity confirmed that strain BGMN1 produces thusin. Both thusin sensitive and resistant strains were detected among clinical isolates of Streptococcus agalactiae. Random mutagenesis of a thusin sensitive strain, S. agalactiae B782, was performed in an attempt to identify the receptor protein for thusin. Three independent thusin resistant mutants were selected and their complete genomes sequenced. Comparative sequence analysis of these mutants with the WT strain revealed that duplication of a region encoding a 79 amino acids repeat in a C-protein α-antigen was a common difference, suggesting it to be responsible for increased resistance to thusin. Since induced thusin resistant mutants showed higher level of resistance than the naturally resistant B761 strain, complete genome sequencing of strain B761 was performed to check the integrity of the C-protein α-antigen-encoding gene. This analysis revealed that this gene is deleted in B761, providing further evidence that this protein promotes interaction of the thusin with receptor.",
publisher = "Springer Science and Business Media Deutschland GmbH",
journal = "Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology",
title = "C-protein α-antigen modulates the lantibiotic thusin resistance in Streptococcus agalactiae",
pages = "1607-1595",
number = "10",
volume = "114",
doi = "10.1007/s10482-021-01626-3"
}

Mirković, N., Obradović, M., O’Connor, P. M., Filipić, B., Jovčić, B., Cotter, P. D.,& Kojić, M.. (2021). C-protein α-antigen modulates the lantibiotic thusin resistance in Streptococcus agalactiae. in Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology
Springer Science and Business Media Deutschland GmbH., 114(10), 1595-1607.
https://doi.org/10.1007/s10482-021-01626-3

Mirković N, Obradović M, O’Connor PM, Filipić B, Jovčić B, Cotter PD, Kojić M. C-protein α-antigen modulates the lantibiotic thusin resistance in Streptococcus agalactiae. in Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology. 2021;114(10):1595-1607.
doi:10.1007/s10482-021-01626-3 .

Mirković, Nemanja, Obradović, Mina, O’Connor, Paula M., Filipić, Brankica, Jovčić, Branko, Cotter, Paul D., Kojić, Milan, "C-protein α-antigen modulates the lantibiotic thusin resistance in Streptococcus agalactiae" in Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology, 114, no. 10 (2021):1595-1607,
https://doi.org/10.1007/s10482-021-01626-3 . .