• The Plant Pathology Journal
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• v.21 no.2
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• pp.119-126
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• 2005

Genomic and phenotypic characteristics of the bacterial strains of Pseudomonas syringae pv. actinidiae and P. syringae pv. syringae collected from several kiwifruit orchards of Korea were investigated and compared with those from Japan to elucidate their phylogenic relationships. All the strains of P. syringae pv. actinidiae and pv. syringae tested were sensitive to copper sulfate but Korean and Japanese strains showed quite different responses to streptomycin. Korean strains were sensitive to streptomycin, but most of the Japanese strains of P. syringae pv. actinidiae were highly resistant to streptomycin. Japanese strains were also relatively more resistant to oxytetracycline than Korean strains. Plasmid profiles were not valuable to distinguish Korean strains of P. syringae pv. actinidiae frombJapanese strains. One or more indigenous plasmids with more than 15 kb in size were detected in all strains of P. syringae pv. actinidiae, but the number and sizes of plasmids harbored in P. syringae pv. actinidiae were variable among the strains regardless of their geographic origins. There also observed no significant relationship among resistance levels of the strains of P. syringae pv. actinidiae to antibiotics, their pathogenicity and plasmid profiles. RAPD profiles were useful to analyze the strains of P. syringae pv. actinidiae and pv. syringae. All the strains of P. syringae pv. actinidiae fell into a wide cluster separated from the strains of P. syringae pv. syringae, but Korean strains of P. syringae pv. actinidiae were separated from Japanese strains. The results support that Korean and Japanese strains of P. syringae pv. actinidiae may have different phylogenic origins.

• Research in Plant Disease
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• v.11 no.1
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• pp.80-84
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• 2005

Bacterial canker of sweet cherry (Prunus avium L.) was observed in farmers\' orchards in Goesan, Chungbuk in 2003. Typical canker symptoms occurred on the branches or twigs of sweet cherry in early spring and bacterial exudates oozed out of the cracked barks of diseased trees. Watersoaked brown symptoms appeared on the leaves and severe infection caused thorough defoliation on the branches or twigs of sweet cherry. When severely infected branches or twigs were cut, irregular and rusty-colored symptoms in sapwood and heartwood were clearly found, indicating that they can serve as specific symptoms of bacterial canker of sweet cherry. The causal bacterium responsible for the symptoms was isolated purely from the infected sapwood of sweet cherry. Based on its morphological, physiological and biochemical characteristics, the causal bacterium was identified as Pseudomonas syringae pv. morsprunorum. The bacterium was pathogenic on sweet cherry and Japanese apricot, but not on peach, cherry, and kiwifruit. It is proposed that the disease be named as bacterial canker of sweet cherry.

• Research in Plant Disease
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• v.24 no.2
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• pp.123-131
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• 2018

Kiwifruit has been cultivated in southern coastal regions of Korea since late 1970s. New cultivars have been successively released in recent years. In this study, we investigated major disease and pest incidences in new kiwifruit cultivars 'Goldone', 'Redvita' and 'Garmrok' at open field in Sacheon for 3 years and rain-proof field in Jeju for 2 years. For the bacterial canker, the 3 new cultivars showed more disease occurrence in Sacheon but rare in Jeju. For leaf spot disease, compared to disease incidence of 20% on 'Hayward' in Sacheon, cv. 'Garmrok' had high incidence about 60% but cv. 'Goldone' and cv. 'Redvita' had low incidence less than 20%. However, in Jeju, diseases incidences of all the new cultivars were lesser than 20%. In the case of Hemiptera, many Halyomorpha halys and Nezara antennata appeared in Sacheon but in Jeju Plautia stali was dominated. Isolated bacterial canker pathogen was identified as Pseudomonas syringae pv. actinidiae biovar 3. Leaf spots pathogens were Phomopsis sp., Phoma sp., Fusarium tricinctum and Alternaria alternata. This study shows the disease information on new kiwifruit cultivars and the adequate disease managements will be required.

• Analytical Science and Technology
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• v.28 no.4
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• pp.299-307
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• 2015

In May 2012, a safety hazard issue arose because some kiwifruit growers in New Zealand had sprayed streptomycin to prevent kiwifruit canker. Therefore, for food safety management, analytical methods to determine streptomycin residues in kiwifruits are required. We developed an analytical method to determine streptomycin residues in kiwifruit samples using liquid chromatograph tandem mass spectrometer (LC-ESI-MS/MS). Streptomycin residues in samples were extracted using 1% formic acid in methanol, centrifugation for 10 min, and subsequent supernatant filtration. Purified samples were subjected to LC-ESI-MS/MS to confirm presence of and quantify streptomycin residues. Average streptomycin recoveries (6 replicates each sample) were in the range of 94.8%-110.6% with relative standard deviations of <10%. The linearity of the concentration range of 0.01-5.0 mg/kg using a matrix-matched calibration gave R² = 0.9995. The limit of quantification (LOQ) was 0.01 mg/kg. Results showed that our analytical method is rapid, simple, and sensitive, with easy sample preparation.

• The Plant Pathology Journal
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• v.35 no.4
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• pp.372-380
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• 2019

Pseudomonas syringae pv. actinidiae (Psa) is by far the most important pathogen of kiwifruit. Sustainable expansion of the kiwifruit industry requires the use of Psa-tolerant or resistant genotypes for the breeding of tolerant cultivars. However, the resistance of most existing kiwifruit cultivars and wild genotypes is poorly understood, and suitable evaluation methods of Psa resistance in Actinidia have not been established. A unique in vitro method to evaluate Psa resistance has been developed with 18 selected Actinidia genotypes. The assay involved debarking and measuring the lesions of cane pieces inoculated with the bacterium in combination with the observation of symptoms such as callus formation, sprouting of buds, and the extent to which Psa invaded xylem. Relative Psa resistance or tolerance was divided into four categories. The division results were consistent with field observations. This is the first report of an in vitro assay capable of large-scale screening of Psa-resistance in Actinidia germplasm with high accuracy and reproducibility. The assay would considerably facilitate the breeding of Psa-resistant cultivars and provide a valuable reference and inspiration for the resistance evaluation of other plants to different pathogens.

• Journal of Life Science
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• v.30 no.1
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• pp.1-9
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• 2020

Pseudomonas syringae pv. actinidiae, the causal agent of a bacterial canker disease in kiwifruit, is subdivided into five genetically distinct populations, namely biovars 1, 2, 3, 5, and 6. Of these, strains belonging to biovar 3 are responsible for a pandemic bacterial canker of kiwifruits since 2008. This study aimed to characterize the structure of the biovar 3 population and investigate the origin of biovar 3 strains isolated in Korea. The genetic variability of fifteen biovar 3 strains, thirteen Korean and two Chinese, were evaluated through random amplified polymorphic DNA (RAPD)-PCR. The RAPD results revealed the presence of eight lineages, designated as subgroups I-VIII, across the biovar 3 strains used in this study. As the strains in subgroups II and III from China were not found in the Korean examples, we concluded that six genetically different biovar 3 subgroups (I, IV, V, VI, VII, and VIII) are present in Korea. In PCR analysis using primers specific to the strains of New Zealand and Europe, Korean strains in subgroups V and VI amplified the relevant DNA bands, suggesting that these were introduced from these two origins, respectively. PCR primers specific to subgroup VIII were developed to monitor the spread of the first biovar 3 strain in Korea, and investigations revealed that this strain was not found in Korea after its first occurrence.

• Research in Plant Disease
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• v.28 no.1
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• pp.26-31
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• 2022

Pseudomonas syringae pv. actinidiae (Psa) is the causal agent responsible for the bacterial canker disease of kiwifruit plants. Psa strains are divided into five different biovars based on genetic and biochemical characteristics. Among them, biovar 2 and 3 strains of Psa were isolated and have been causing widespread damages in Korea. One of the most effective ways to control Psa is to use an antibiotic such as streptomycin. However, Psa strains resistant to this antibiotic were isolated in Korea, and an earlier study revealed that the resistance in the biovar 2 is associated with strA-strB genes. This study aimed to determine the molecular resistance mechanism of Psa biovar 3 strains to streptomycin. Sequencing the rpsL gene encoding ribosomal protein S12 from three streptomycin-resistant strains screened in the laboratory revealed that a spontaneous mutation occurred either at codon 43 or 88. Meanwhile, in four streptomycin-resistant strains of Psa biovar 3 isolated from two kiwifruit orchards, a single nucleotide in codon 43 of the rpsL, which is AAA in streptomycin-sensitive strain, was substituted for AGA causing an amino acid change from lysine to arginine. The resistant mechanism in all biovar 3 strains obtained in Korea was identified as a mutation of the rpsL gene.

• Journal of Plant Biotechnology
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• v.42 no.4
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• pp.342-349
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• 2015

Kiwifruit is a new fruit crop that was commercialized in the late 1970s. Recently, its cultivation and consumption have increased rapidly worldwide. Kiwifruit is a dioecious, deciduous, and climbing plant having fruit with hairs and various flesh colors and a variation in ploidy level; however, the industry consists of very simple cultivars or genotypes. The need for efficient cultivar improvement together with the evolutional and biological perspectives based on unique plant characteristics, have recently encouraged genome analysis and bioinformatics application. The draft genome sequence and chloroplast genome sequence of kiwifruit were released in 2013 and 2015, respectively; and gene annotation has been in progress. Recently, transcriptome analysis has shifted from previous ESTs analysis to the RNA-seq platform for intensive exploration of controlled genetic expression and gene discovery involved in fruit ascorbic acid biosynthesis, flesh coloration, maturation, and vine bacterial canker tolerance. For improving conventional breeding efficiency, molecular marker development and genetic linkage map construction have advanced from basic approaches using RFLP, RAPD, and AFLP to the development of NGS-based SSR and SNP markers linked to agronomically important traits and the construction of highly saturated linkage maps. However, genome and transcriptome studies have been limited in Korea. In the near future, kiwifruit genome and transcriptome studies are expected to translate to the practical application of molecular breeding.

• 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.

• 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.