TY  - 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 . .

TY  - JOUR
AU  - Ivanović, Milan
AU  - Kuzmanović, Nemanja
AU  - Prokić, Anđelka
AU  - Blagojević, Nevena
AU  - Obradović, Aleksa
AU  - Gašić, Katarina
PY  - 2014
UR  - https://plantarum.izbis.bg.ac.rs/handle/123456789/1104
AB  - In this study, three bacterial DNA extraction procedures were compared prior to real-time PCR. Healthy pear leaves and twigs were crushed in antioxidant maceration buffer and spiked with Erwinia amylovora to final concentrations from 2.1 x 10(6) to 2.1 x 10(1) cells ml(-1). Bacterial DNA was extracted from aliquots of spiked crude extracts using (i) isopropanol, (ii) REDExtract-N-Amp (TM) Plant PCR kit, and (iii) Taylor's modified DNA purification procedure. The ams region of the chromosomal DNA was selected as target for the real-time PCR. In this study, the REDExtract-N-Amp (TM) and Taylor's modified DNA extraction procedure were most successful in removing PCR inhibitors, leading to detection of 2.1x10(2) E. amylovora CFU/ml. At this concentration, pathogen can be efficiently detected in less than 5 h in spite of inhibitors and plant DNA reducing sensitivity of the reaction. These two methods increased amplification efficiency in real-time PCR compared to a simple isopropanol DNA extraction procedure from plant tissues, where the lowest detected concentration was 2.1 x 10(4) CFU/ml. In our research, real-time PCR has proven to be very sensitive method for detection of E. amylovora in plant material. It was 100 times more sensitive compared to other conventional PCR procedures.
PB  - International Society for Horticultural Science
T2  - Acta Horticulturae
T1  - Evaluation of Three Extraction Methods for Detection of Erwinia amylovora from Pear Leaves by Real-Time PCR
EP  - 84
SP  - 81
VL  - 1056
DO  - 10.17660/ActaHortic.2014.1056.10
ER  - 

@article{
author = "Ivanović, Milan and Kuzmanović, Nemanja and Prokić, Anđelka and Blagojević, Nevena and Obradović, Aleksa and Gašić, Katarina",
year = "2014",
abstract = "In this study, three bacterial DNA extraction procedures were compared prior to real-time PCR. Healthy pear leaves and twigs were crushed in antioxidant maceration buffer and spiked with Erwinia amylovora to final concentrations from 2.1 x 10(6) to 2.1 x 10(1) cells ml(-1). Bacterial DNA was extracted from aliquots of spiked crude extracts using (i) isopropanol, (ii) REDExtract-N-Amp (TM) Plant PCR kit, and (iii) Taylor's modified DNA purification procedure. The ams region of the chromosomal DNA was selected as target for the real-time PCR. In this study, the REDExtract-N-Amp (TM) and Taylor's modified DNA extraction procedure were most successful in removing PCR inhibitors, leading to detection of 2.1x10(2) E. amylovora CFU/ml. At this concentration, pathogen can be efficiently detected in less than 5 h in spite of inhibitors and plant DNA reducing sensitivity of the reaction. These two methods increased amplification efficiency in real-time PCR compared to a simple isopropanol DNA extraction procedure from plant tissues, where the lowest detected concentration was 2.1 x 10(4) CFU/ml. In our research, real-time PCR has proven to be very sensitive method for detection of E. amylovora in plant material. It was 100 times more sensitive compared to other conventional PCR procedures.",
publisher = "International Society for Horticultural Science",
journal = "Acta Horticulturae",
title = "Evaluation of Three Extraction Methods for Detection of Erwinia amylovora from Pear Leaves by Real-Time PCR",
pages = "84-81",
volume = "1056",
doi = "10.17660/ActaHortic.2014.1056.10"
}

Ivanović, M., Kuzmanović, N., Prokić, A., Blagojević, N., Obradović, A.,& Gašić, K.. (2014). Evaluation of Three Extraction Methods for Detection of Erwinia amylovora from Pear Leaves by Real-Time PCR. in Acta Horticulturae
International Society for Horticultural Science., 1056, 81-84.
https://doi.org/10.17660/ActaHortic.2014.1056.10

Ivanović M, Kuzmanović N, Prokić A, Blagojević N, Obradović A, Gašić K. Evaluation of Three Extraction Methods for Detection of Erwinia amylovora from Pear Leaves by Real-Time PCR. in Acta Horticulturae. 2014;1056:81-84.
doi:10.17660/ActaHortic.2014.1056.10 .

Ivanović, Milan, Kuzmanović, Nemanja, Prokić, Anđelka, Blagojević, Nevena, Obradović, Aleksa, Gašić, Katarina, "Evaluation of Three Extraction Methods for Detection of Erwinia amylovora from Pear Leaves by Real-Time PCR" in Acta Horticulturae, 1056 (2014):81-84,
https://doi.org/10.17660/ActaHortic.2014.1056.10 . .