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Characterization of Pseudomonas syringae Strains Associated with Shoot Blight of Raspberry and Blackberry in Serbia

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Authors
Ivanović, Milan
Prokić, Anđelka
Gašić, Katarina
Menković, Jelena
Kuzmanović, Nemanja
Zlatković, Nevena
Obradović, Aleksa
Article (Published version)
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Abstract
During May 2016, severe blight symptoms were observed in several raspberry and blackberry fields in Serbia. In total, 22 strains were isolated: 16 from symptomatic raspberry shoots, 2 from asymptomatic raspberry leaves, and 4 from symptomatic blackberry shoots. Additionally, eight raspberry strains, isolated earlier from two similar outbreaks, were included in the study. Pathogenicity of the strains was confirmed on detached raspberry and blackberry shoots by reproducing the symptoms of natural infection. The strains were Gram-negative, fluorescent on King’s medium B, ice nucleation positive, and utilized glucose oxidatively. All strains were levan positive, oxidase negative, nonpectolytic, arginine dihydrolase negative, and induced hypersensitivity in tobacco leaves (LOPAT + −−− +, Pseudomonas group Ia). Furthermore, all strains liquefied gelatin and hydrolyzed aesculin but did not show tyrosinase activity or utilize tartrate (GATTa + + − −). Tentative identification using morphology,... LOPAT, GATTa, and ice-nucleating ability tests suggested that isolated strains belong to Pseudomonas syringae. The syrB gene associated with syringomycin production was detected in all strains. DNA fingerprints with REP, ERIC, and BOX primers generated identical profiles for 29 strains, except for strain KBI 222, which showed a unique genomic fingerprint. In all, 9 of 10 selected strains exhibited identical sequences of four housekeeping genes: gyrB, rpoD, gapA, and gltA. Five nucleotide polymorphisms were found in strain KBI 222 at the rpoD gene locus only. In the phylogenetic tree based on a concatenated sequence of all four housekeeping genes, strains clustered within phylogroup 2 (i.e., genomospecies 1) of the P. syringae species complex, with pathotype strains of P. syringae pv. aceris and P. syringae pv. solidagae as their closest relatives. There was no correlation between genotype and geographic origin, particular outbreak, host, or cultivar.

Keywords:
epidemiology / genetic relatedness / multilocus sequence analysis / Pseudomonas blight / repetitive sequence-based PCR
Source:
PLANT DISEASE, 2023, 107, 3
Publisher:
  • APS Publications
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)

DOI: 10.1094/PDIS-06-22-1425-RE

ISSN: 0191-2917; 1943-7692

Scopus: 2-s2.0-85152105647
[ Google Scholar ]
URI
https://plantarum.izbis.bg.ac.rs/handle/123456789/715
Collections
  • Radovi istraživača / Researchers' publications
Institution/Community
IZBIS
TY  - JOUR
AU  - Ivanović, Milan
AU  - Prokić, Anđelka
AU  - Gašić, Katarina
AU  - Menković, Jelena
AU  - Kuzmanović, Nemanja
AU  - Zlatković, Nevena
AU  - Obradović, Aleksa
PY  - 2023
UR  - https://plantarum.izbis.bg.ac.rs/handle/123456789/715
AB  - During May 2016, severe blight symptoms were observed in several raspberry and blackberry fields in Serbia. In total, 22 strains were isolated: 16 from symptomatic raspberry shoots, 2 from asymptomatic raspberry leaves, and 4 from symptomatic blackberry shoots. Additionally, eight raspberry strains, isolated earlier from two similar outbreaks, were included in the study. Pathogenicity of the strains was confirmed on detached raspberry and blackberry shoots by reproducing the symptoms of natural infection. The strains were Gram-negative, fluorescent on King’s medium B, ice nucleation positive, and utilized glucose oxidatively. All strains were levan positive, oxidase negative, nonpectolytic, arginine dihydrolase negative, and induced hypersensitivity in tobacco leaves (LOPAT + −−− +, Pseudomonas group Ia). Furthermore, all strains liquefied gelatin and hydrolyzed aesculin but did not show tyrosinase activity or utilize tartrate (GATTa + + − −). Tentative identification using morphology, LOPAT, GATTa, and ice-nucleating ability tests suggested that isolated strains belong to Pseudomonas syringae. The syrB gene associated with syringomycin production was detected in all strains. DNA fingerprints with REP, ERIC, and BOX primers generated identical profiles for 29 strains, except for strain KBI 222, which showed a unique genomic fingerprint. In all, 9 of 10 selected strains exhibited identical sequences of four housekeeping genes: gyrB, rpoD, gapA, and gltA. Five nucleotide polymorphisms were found in strain KBI 222 at the rpoD gene locus only. In the phylogenetic tree based on a concatenated sequence of all four housekeeping genes, strains clustered within phylogroup 2 (i.e., genomospecies 1) of the P. syringae species complex, with pathotype strains of P. syringae pv. aceris and P. syringae pv. solidagae as their closest relatives. There was no correlation between genotype and geographic origin, particular outbreak, host, or cultivar.
PB  - APS Publications
T2  - PLANT DISEASE
T1  - Characterization of Pseudomonas syringae Strains Associated with Shoot Blight of Raspberry and Blackberry in Serbia
IS  - 3
VL  - 107
DO  - 10.1094/PDIS-06-22-1425-RE
ER  - 
@article{
author = "Ivanović, Milan and Prokić, Anđelka and Gašić, Katarina and Menković, Jelena and Kuzmanović, Nemanja and Zlatković, Nevena and Obradović, Aleksa",
year = "2023",
abstract = "During May 2016, severe blight symptoms were observed in several raspberry and blackberry fields in Serbia. In total, 22 strains were isolated: 16 from symptomatic raspberry shoots, 2 from asymptomatic raspberry leaves, and 4 from symptomatic blackberry shoots. Additionally, eight raspberry strains, isolated earlier from two similar outbreaks, were included in the study. Pathogenicity of the strains was confirmed on detached raspberry and blackberry shoots by reproducing the symptoms of natural infection. The strains were Gram-negative, fluorescent on King’s medium B, ice nucleation positive, and utilized glucose oxidatively. All strains were levan positive, oxidase negative, nonpectolytic, arginine dihydrolase negative, and induced hypersensitivity in tobacco leaves (LOPAT + −−− +, Pseudomonas group Ia). Furthermore, all strains liquefied gelatin and hydrolyzed aesculin but did not show tyrosinase activity or utilize tartrate (GATTa + + − −). Tentative identification using morphology, LOPAT, GATTa, and ice-nucleating ability tests suggested that isolated strains belong to Pseudomonas syringae. The syrB gene associated with syringomycin production was detected in all strains. DNA fingerprints with REP, ERIC, and BOX primers generated identical profiles for 29 strains, except for strain KBI 222, which showed a unique genomic fingerprint. In all, 9 of 10 selected strains exhibited identical sequences of four housekeeping genes: gyrB, rpoD, gapA, and gltA. Five nucleotide polymorphisms were found in strain KBI 222 at the rpoD gene locus only. In the phylogenetic tree based on a concatenated sequence of all four housekeeping genes, strains clustered within phylogroup 2 (i.e., genomospecies 1) of the P. syringae species complex, with pathotype strains of P. syringae pv. aceris and P. syringae pv. solidagae as their closest relatives. There was no correlation between genotype and geographic origin, particular outbreak, host, or cultivar.",
publisher = "APS Publications",
journal = "PLANT DISEASE",
title = "Characterization of Pseudomonas syringae Strains Associated with Shoot Blight of Raspberry and Blackberry in Serbia",
number = "3",
volume = "107",
doi = "10.1094/PDIS-06-22-1425-RE"
}
Ivanović, M., Prokić, A., Gašić, K., Menković, J., Kuzmanović, N., Zlatković, N.,& Obradović, A.. (2023). Characterization of Pseudomonas syringae Strains Associated with Shoot Blight of Raspberry and Blackberry in Serbia. in PLANT DISEASE
APS Publications., 107(3).
https://doi.org/10.1094/PDIS-06-22-1425-RE
Ivanović M, Prokić A, Gašić K, Menković J, Kuzmanović N, Zlatković N, Obradović A. Characterization of Pseudomonas syringae Strains Associated with Shoot Blight of Raspberry and Blackberry in Serbia. in PLANT DISEASE. 2023;107(3).
doi:10.1094/PDIS-06-22-1425-RE .
Ivanović, Milan, Prokić, Anđelka, Gašić, Katarina, Menković, Jelena, Kuzmanović, Nemanja, Zlatković, Nevena, Obradović, Aleksa, "Characterization of Pseudomonas syringae Strains Associated with Shoot Blight of Raspberry and Blackberry in Serbia" in PLANT DISEASE, 107, no. 3 (2023),
https://doi.org/10.1094/PDIS-06-22-1425-RE . .

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