TY  - JOUR
AU  - Zlatković, Nevena
AU  - Gašić, Katarina
AU  - Kuzmanović, Nemanja
AU  - Prokić, Anđelka
AU  - Ivanović, Milan
AU  - Zivkovic, Svetlana
AU  - Obradović, Aleksa
PY  - 2022
UR  - https://plantarum.izbis.bg.ac.rs/handle/123456789/744
AB  - Acidovorax citrulli, the causal agent of bacterial fruit blotch (BFB), is an economically
important pathogen of watermelon and related plant species worldwide. In the period 2014–2018,
several outbreaks of BFB were observed in major watermelon production regions in Serbia. A total of
43 strains, isolated from symptomatic watermelon tissue, were analyzed by biochemical, pathogenic
and molecular tests. Based on the phenotypic characteristics, PCR assay, and 16S rRNA gene
sequence analysis, all strains were identified as A. citrulli. A multilocus sequence analysis of the four
housekeeping (adk, gyrB, pilT and gltA) and three virulence genes (Aave_1548, avrRx01 and luxR)
revealed that studied A. citrulli strains represent the homogeneous population and they clustered
together with group II reference strain AAC00-1. Strain affiliation to group II was confirmed by PCR
based on the putative type III secretion effector gene and by duplex PCR test. High homogeneity
of studied strains was also confirmed by BOX-PCR. Differences were observed for two strains in
their pathogenicity as well as susceptibility to copper compounds. Moreover, six major watermelon
varieties grown in Serbia showed high sensitivity to the pathogen, while cross inoculation assay
revealed that the strains were able to infect other species within the Cucurbitaceae family.
PB  - MDPI
T2  - Agronomy
T1  - Polyphasic Characterization of Acidovorax citrulli Strains Originating from Serbia
SP  - 235
VL  - 12
VL  - 2
DO  - 10.3390/agronomy12020235
ER  - 

@article{
author = "Zlatković, Nevena and Gašić, Katarina and Kuzmanović, Nemanja and Prokić, Anđelka and Ivanović, Milan and Zivkovic, Svetlana and Obradović, Aleksa",
year = "2022",
abstract = "Acidovorax citrulli, the causal agent of bacterial fruit blotch (BFB), is an economically
important pathogen of watermelon and related plant species worldwide. In the period 2014–2018,
several outbreaks of BFB were observed in major watermelon production regions in Serbia. A total of
43 strains, isolated from symptomatic watermelon tissue, were analyzed by biochemical, pathogenic
and molecular tests. Based on the phenotypic characteristics, PCR assay, and 16S rRNA gene
sequence analysis, all strains were identified as A. citrulli. A multilocus sequence analysis of the four
housekeeping (adk, gyrB, pilT and gltA) and three virulence genes (Aave_1548, avrRx01 and luxR)
revealed that studied A. citrulli strains represent the homogeneous population and they clustered
together with group II reference strain AAC00-1. Strain affiliation to group II was confirmed by PCR
based on the putative type III secretion effector gene and by duplex PCR test. High homogeneity
of studied strains was also confirmed by BOX-PCR. Differences were observed for two strains in
their pathogenicity as well as susceptibility to copper compounds. Moreover, six major watermelon
varieties grown in Serbia showed high sensitivity to the pathogen, while cross inoculation assay
revealed that the strains were able to infect other species within the Cucurbitaceae family.",
publisher = "MDPI",
journal = "Agronomy",
title = "Polyphasic Characterization of Acidovorax citrulli Strains Originating from Serbia",
pages = "235",
volume = "12, 2",
doi = "10.3390/agronomy12020235"
}

Zlatković, N., Gašić, K., Kuzmanović, N., Prokić, A., Ivanović, M., Zivkovic, S.,& Obradović, A.. (2022). Polyphasic Characterization of Acidovorax citrulli Strains Originating from Serbia. in Agronomy
MDPI., 12, 235.
https://doi.org/10.3390/agronomy12020235

Zlatković N, Gašić K, Kuzmanović N, Prokić A, Ivanović M, Zivkovic S, Obradović A. Polyphasic Characterization of Acidovorax citrulli Strains Originating from Serbia. in Agronomy. 2022;12:235.
doi:10.3390/agronomy12020235 .

Zlatković, Nevena, Gašić, Katarina, Kuzmanović, Nemanja, Prokić, Anđelka, Ivanović, Milan, Zivkovic, Svetlana, Obradović, Aleksa, "Polyphasic Characterization of Acidovorax citrulli Strains Originating from Serbia" in Agronomy, 12 (2022):235,
https://doi.org/10.3390/agronomy12020235 . .

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 . .