• Research in Plant Disease
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• v.9 no.3
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• pp.116-120
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• 2003

A PCR method that combines biological and enzymatic amplification of PCR targets was developed for the detection of Pseudomonas syringae pv. actinidiae on kiwifruit leaves. A nested PCR was performed with primers designes from the coding sequence of the cfl gene, which is involved in production of the phytotoxin coronatine. The first and second primer sets efficiently amplified expected 665 and 310-bp fragments, respectively. With two successive amplifications, as few as 20 CFU/ml of P. syringae pv. actinidiae could be detected on ethidium bromide-stained agarose gel. Leaf samples were collected from 4 kiwifruit trees showing yellow halo spots on leaves and incubated in pepton-sucrose broth for 12 h at $16^{\circ}$C before PCR amplification. Positive detection was obtained with one sample, which was proved as a diseased plant in the next spring.

• Journal of Microbiology
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• v.41 no.1
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• pp.16-21
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• 2003

We have collected eight high-level streptomycin-resistant strains of Pseudomonas marginalis and P. syringae pv. actinidiae which were isolated from kiwifruit orchards in Korea and Japan, The molecular mechanisms of resistance were investigated by the PCR, susceptibility tests, and nucleotide sequence analysis. Of the eight high-level streptomycin-resistant strains, four harbored strA-strB genes, which encode streptomycin-inactivating enzymes. While the three Korean strains of R marginalis did not have plasmid and carried the resistant genes in the chromosomes, the Japanese strain of P. syringae pv. actinidiae had a plasmid containing strA-strB genes. The myomycin susceptibility test demonstrated that the high-level resistance to streptomycin of the remaining four strains is associated with mutations in the rpsL gene. Nucleotide sequence analyses revealed that they contain a single base-pair mutation in codon 43 of their rpsL gene.

• The Plant Pathology Journal
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• v.33 no.1
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• pp.21-33
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• 2017

Citrus blast caused by bacterium Pseudomonas syringae is a very important disease of citrus occuring in many areas of the world, but with few data about genetic structure of the pathogen involved. Considering the above fact, this study reports genetic characterization of 43 P. syringae isolates obtained from plant tissue displaying citrus blast symptoms on mandarin (Citrus reticulata) in Montenegro, using multilocus sequence analysis of gyrB, rpoD, and gap1 gene sequences. Gene sequences from a collection of 54 reference pathotype strains of P. syringae from the Plant Associated and Environmental Microbes Database (PAMDB) was used to establish a genetic relationship with our isolates obtained from mandarin. Phylogenetic analyses of gyrB, rpoD, and gap1 gene sequences showed that P. syringae pv. syringae causes citrus blast in mandarin in Montenegro, and belongs to genomospecies 1. Genetic homogeneity of isolates suggested that the Montenegrian population might be clonal which indicates a possible common source of infection. These findings may assist in further epidemiological studies of this pathogen and for determining mandarin breeding strategies for P. syringae control.

• Research in Plant Disease
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• v.9 no.4
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• pp.213-216
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• 2003

A bacterial disease of tea plants(Camellia sinensis L.) was found in the graftage nursery grown under vinyl house conditions in Suncheon city, Korea, in spring of 2002. The primary symptoms of the disease include small, water-soaked and dark brown spot development on the young leaves. This spot gradually increases in size, especially taking on elongate shape along the midrib or vein of the leaf, and then turns black. The diseased leaves were defoliated easily. Ten strains were isolated from the infected leaf. Inoculation on tea leaf with these isolates produced the same symptoms of naturally infected plants. On the basis of stain reactions, morphological characterization, colony pattern, physiological and biochemical reactions, the bacterium was identified as Pseudomonas syringae pv. theae. This is the first report of brown blight of tea plant in Korea.

• Journal of Life Science
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• v.25 no.2
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• pp.136-150
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• 2015

Pseudomonas syringae pathovar tabaci is a plant pathogenic bacterium that causes wildfire disease in tobacco plants. In P. syringae pv. tabaci, PsyI, a LuxI-type protein, acts as an AHL synthase, while primary and secondary sequence analysis of PsyR has revealed that it is a homolog of the LuxR-type transcriptional regulator that responds to AHL molecules. In this study, using phenotypic and genetic analyses in P. syringae pv. tabaci, we show the effect of PsyR protein as a quorum-sensing (QS) transcriptional regulator. Regulatory effects of PsyR on swarming motility and production of siderophores, tabtoxin, and N-acyl homoserine lactones were examined via phenotypic assays, and confirmed by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR). Further qRT-PCR showed that PsyR regulates expression of these virulence genes in response to environmental signals. However, an upstream region of the gene was not bound with purified MBP-PsyR protein; rather, PsyR was only able to shift the upstream region of psyI. These results suggested that PsyR may be indirectly controlled via intermediate-regulatory systems and that auto-regulation by PsyR does not occur.

• The Korean Journal of Pesticide Science
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• v.19 no.2
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• pp.119-124
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• 2015

Bacterial leaf spot, caused by Pseudomonas syringae pv. syrinage, is a very damaging disease to green pumpkin in Gong-ju and Non-san nursery. However, there is no good method to control the disease in Korea. Growth inhibition of pathogen on medium, control efficacy on seedling stage, and seed treatment effect of 6 anti-bacterial pesticides were investigated for selection of the best pesticide for seed treatment and control of the disease. Growth inhibition zone on King's B medium were the largest by oxytetracycline 170 ppm and oxytetracycline 15 ppm + streptomycin sulfate 188 ppm, oxolinic acid 200 ppm, streptomycin 200 ppm were next respectively. Control efficacy of oxytetracycline 1.5% + streptomycin sulfate 18.8% WP and oxytetracycline 17% WP on seedling stage were 71.4% and 49.4%, respectively. Seed treatment of oxytetracycline 15 ppm + streptomycin sulfate 188 ppm on the artificially inoculated seeds inhibits pathogen growth completely from the treated seeds and 96% control efficacy on grow-out test of the treated seeds. Seed treatment of streptomycin 100 ppm (2,000 dilution of streptomycin 20%) on the artificially inoculated seeds allow 280 cfu/g of pathogen growth from the treated seeds and 60% control efficacy on grow-out test of the treated seeds. Seed treatment of oxytetracycline 85 ppm (2000 dilution of oxytetracycline 17% WP) on the artificially inoculated seeds allow 80 cfu/g of pathogen growth from the treated seeds and 90% control efficacy on grow-out test of the treated seeds. These results suggested that oxytetracycline 1.5% + streptomycin sulfate 18.8% WP was the best pesticide for seed treatment to control of the bacterial spot disease by Pseudomonas syringae pv. syringae.

• Korean Journal of Microbiology
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• v.50 no.3
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• pp.245-248
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• 2014

Pseudomonas syringae pv. actinidiae is the causal agent of bacterial canker in kiwifruit (genus Actinidia). Multilocus sequence analysis of seven housekeeping and 11 type III effector genes differentiated the virulent P. syringae pv. actinidiae isolates worldwide into three groups designated as Psa1-Psa3. In this work, a total of 12 P. syringae pv. Actinidiae strains, including three Psa1, three Psa2, three Psa3 strains isolated from Korea and three Psa3 strains from Italy, were compared based on their phenotypic properties. Strains with different geographic origins had unique growth patterns as demonstrated by growth rate at several temperatures; all tested strains exhibited maximum growth at temperatures below $22^{\circ}C$, while the growth of Psa3 strains was completely inhibited above $30^{\circ}C$. Psa3 strains isolated from Korea had longer lag phases than the Psa3 strains from Italy. The Psa2 strains were different from Psa1 and Psa3 strains in the API 20NE test, in which the Psa2 strains could not utilize potassium gluconate, capric acid and trisodium citrate. Psa3 strains isolated from Korea could hydrolyze esculin. The API ZYM test showed that ${\beta}$-glucosidase activity was detected only from Psa3 strains. The strains belonging to the three Psa groups differed with regard to their susceptibility to ampicillin, novobiocin, and oleandomycin.

• The Plant Pathology Journal
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• v.28 no.3
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• pp.295-298
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• 2012

Genetic diversity among Pseudomonas syringae pv. morsprunorum isolates from Prunus mume in Korea and Japan was investigated by comparative sequence analysis of the 16S rRNA gene. The strains included 24 field isolates recovered from P. mume in Korea along with seven Japanese strains. Two strains isolated from P. salicina in Japan, one strain from P. avium in the United Kingdom, and the pathotype strain were also used for comparison with their 16S rRNA gene sequences. Nearly complete 16S rRNA gene sequences were sequenced in all 35 strains, and three sequence types, designated types I, II and III, were identified. Eleven strains consisting of five Korean isolates, five Japanese strains, and one strain from the United Kingdom belonged to type I, whereas the pathotype strain and another 19 Korean isolates belonged to type III. Another four Japanese strains belonged to type II. Type I showed 98.9% sequence homology with type III. Type I and II had only two heterogeneous bases. The 16S rRNA sequence types were correlated with the races of P. syringae pv. morsprunorum. Type I and II strains belonged to race 1, whereas type III isolates were included in race 2. Sequence analyses of the 16S rRNA gene from P. syringae pv. morsprunorum were useful in identifying the races and can further be used for epidemiological surveillance of this pathogen.

• The Plant Pathology Journal
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• v.26 no.4
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• pp.402-408
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• 2010

Plant matrix metalloproteases (MMPs) are a family of apoplastic metalloproteases closely related to human matrilysins. Up-regulation of Nicotiana benthamiana matrix metalloprotease 1 (NMMP1) expression by treatment with pathogens, ethephon and aging indicates that the gene is related to plant defense and the aging process through ethylene signaling. NMMP1 expression was higher than in normal growth leaves following infection with an incompatible pathogen Pseudomonas syringae pv. tomato T1 or a compatible pathogen P. syringae pv. tabaci and in aged leaves. Transient overexpression of NMMP1 in N. benthamiana leaves lowered the growth of P. syringae pv. tabaci. However, NMMP1-silenced leaves showed increased growth of P. syringae pv. tabaci. These data strongly suggest that NMMP1 in N. benthamiana is a defense related gene, which is positively regulated by ethylene.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.385-393
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• 2016

Pseudomonas syringae pv. actinidiae causes bacterial canker disease in kiwifruit. Owing to the prohibition of agricultural antibiotic use in major kiwifruit-cultivating countries, alternative methods need to be developed to manage this disease. Bacteriophages are viruses that specifically infect target bacteria and have recently been reconsidered as potential biological control agents for bacterial pathogens owing to their specificity in terms of host range. In this study, we isolated bacteriophages against P. syringae pv. actinidiae from soils collected from kiwifruit orchards in Korea and selected seven bacteriophages for further characterization based on restriction enzyme digestion patterns of genomic DNA. Among the studied bacteriophages, two belong to the Myoviridae family and three belong to the Podoviridae family, based on morphology observed by transmission electron microscopy. The host range of the selected bacteriophages was confirmed using 18 strains of P. syringae pv. actinidiae, including the Psa2 and Psa3 groups, and some were also effective against other P. syringae pathovars. Lytic activity of the selected bacteriophages was sustained in vitro until 80 h, and their activity remained stable up to 50℃, at pH 11, and under UV-B light. These results indicate that the isolated bacteriophages are specific to P. syringae species and are resistant to various environmental factors, implying their potential use in control of bacterial canker disease in kiwifruits.