• The Plant Pathology Journal
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• v.39 no.1
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• pp.88-107
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• 2023

In the present investigation, bacterial isolates from infected apple trees causing apple canker during winter were studied in the northern Gyeongbuk Province, Korea. The pathogen was identified as Pseudomonas syringae pv. syringae (Pss) through various physiological and biochemical characterization assays such as BIOLOG, gas chromatography of fatty acid methyl esters, and 16S rRNA. Bioassays for the production of phytotoxins were positive for syringopeptin and syringomycin against Bacillus megaterium and Geotrichum candidum, respectively. The polymerase chain reaction (PCR) method enabled the detection of toxin-producing genes, syrB1, and sypB in Pss. The differentiation of strains was performed using LOPAT and GATTa tests. Pss further exhibited ice nucleation activity (INA) at a temperature of -0.7℃, indicating an INA⁺ bacterium. The ice-nucleating temperature was -4.7℃ for a non-treated control (sterilized distilled water), whereas it was -9.6℃ for an INA^- bacterium Escherichia coli TOP10. These methods detected pathogenic strains from apple orchards. Pss might exist in an apple tree during ice injury, and it secretes a toxin that makes leaves yellow and cause canker symptoms. Until now, Korea has not developed antibiotics targeting Pss. Therefore, it is necessary to develop effective disease control to combat Pss in apple orchards. Pathogenicity test on apple leaves and stems showed canker symptoms. The pathogenic bacterium was re-isolated from symptomatic plant tissue and confirmed as original isolates by 16S rRNA. Repetitive element sequence-based PCR and enterobacterial repetitive intergenic consensus PCR primers revealed different genetic profiles within P. syringae pathovars. High antibiotic susceptibility results showed the misreading of mRNA caused by streptomycin and oxytetracycline.

• Research in Plant Disease
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• v.19 no.2
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• pp.102-107
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• 2013

The percent of diseased fruit by Diaporthe citri was the most 16.8% in 2010 and the least 3.8% in 2005, and the average was 8.8% from 2003 to 2012. The degrees of disease incidence were relatively high, 17.0, 22.6, 19.2 and 18.9%, in 2003, 2004, 2007 and 2010, respectively. The percent of diseased fruit by Elsinoe fawcettii was the most 0.72% in 2004, and then gradually decreased to 0.08% in 2008, and the average was from 0.28% from 2004 to 2011. In case of canker by Xanthomonas citri subsp. citri, the percent of diseased fruit was the most 2.19% in 2004, and gradually decreased to 0.08 and 0.10% in 2009 and 2010, respectively. The average was 0.81% from 2004 to 2012. The melanose by D. citri was most in south-east area of Jeju Island, and east and west in case of canker. The melanose symptoms were initially appeared from mid June, mainly peaked late of July to late August. The canker symptoms were initially produced on leaves of natsudaidai and grapefruit cultivated in open field middle of June in 2010 and 2011 and late of May in 2012. The scab disease was firstly appeared approx. early or mid May and then rapidly increased late of May to middle of June. The inoculum was mainly produced from dead twigs late June to early of August.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.124-124
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• 2003

Citrus canker is very important disease in international trade of citrus. The disease was usually take place from late of June, and severe middle of July to middle of August, though disease occurrence was affected by environmental conditions. In pathogenicity test, three varieties, orange, lemon and kiyomi among 7 varieties, were succeptible, two varieties, satsuma mandarin and iwasachi, intermediate resistant. On the other hand, shiranuhi and yuzu were resistant relatively. The pathogen, Xanthomonas axonopodis pv. citri, grew well in PD broth adjusted to pH 7.0 at 26$^{\circ}C$. It's growth was best in medium containing group of monosaccharide as a carbon source and group of ammonium as a nitrogen source. Tow isolates were resistant to streptomycin among 11 isolates isolated from diseased leaves in field in Jeju-Do. The streptomycin sensitives isolate was controlled by in greenhouse test. On the other hand, the resistant and sensitive isolates were controlled by treatment with copper sulfate, the control value is 88.7％ and 90.6％, respectively.

• Research in Plant Disease
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• v.18 no.2
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• pp.129-132
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• 2012

Citrus bacterial canker is an economically important disease affecting citrus production in many citrusgrowing areas and several pathotypes have been recognized within the Xanthomonas pathogens causing canker. In view of the containment of the disease, accurate identification of the causal bacterium is important. In this study, triplex PCR method was developed by using the previously reported primers. Two groups of primer combination, such as, one group including primers 2/3, J-pth1/J-pth2 and XACF/XACR, and another group 2/3, J-pth1/J-pth2 and Xac01/Xac02, were suitable for the detection and differentiation of X. a. pv. citri $A^w$, X. a. pv. aurantifolii B and C, and X. a. pv. citrumelo E strains. Moreover, the primer combination of Xac01 and J-pth2 promised us to use as a specific primer set to detect X. a. pv. citrumelo E strain. The PCR methods developed in this study could be used for the rapid differentiation of Xanthomonas pathotypes of citrus.

• Research in Plant Disease
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• v.8 no.4
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• pp.250-253
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• 2002

This research was conducted to develop a simple and effective method for pathogenicity assay of the causal agent of bacterial canker on kiwifruit. The developed method is a modified version of syringe-infiltration method that is used in the assay fer the hypersensitive response assay. Bacterial cell suspensions in 50 mM potassium phosphate buffer(pH 7.5) were infiltrated using a plastic syringe with 25G needle into primary leaves of five-year-old kiwifruit. Typical symptoms of bacterial canker were observed five days after infiltration. Symptoms developed on the leaves were detected in these inocula that treated above 10$^4$cfu/ml or above. Using this technique, host range of Pseudomonas syringae pv. actinidiae and three other plant pathogenic pseudomonads were investigated for 25 different plant species. The various symptoms were showed depend-ing on different plant species and inoculated pathogen combinations. This method has the advantage that symptoms can be showed faster compared to other methods and high humid conditions are not required.

• Research in Plant Disease
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• v.26 no.1
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• pp.44-47
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• 2020

Streptomycin resistant isolates of Pseudomonas syringae pv. actinidiae, the causal agent of bacterial canker in kiwifruit, were found in Korea. A total of 734 isolates of P. syringae pv. actinidiae collected between 2008 and 2017 from bacterial canker infections in 111 kiwifruit orchards were assessed for streptomycin resistance. The survival of each isolate was screened against 100 ㎍/ml of streptomycin. Among 734 isolates, 38 streptomycin resistant P. syringae pv. actinidiae isolates originated from nine orchards were found. Streptomycin resistant isolates belonging to biovar 2 were found in several individual years, but ones belonging to biovar 3 were found in Korea only since 2016. Therefore, to use streptomycin for control of bacterial canker in kiwifruit orchards should be very careful, and it is necessary to check the streptomycin susceptibility of the pathogen before use in kiwifruit orchards.

• Research in Plant Disease
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• v.23 no.1
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• pp.35-41
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• 2017

Pseudomonas syringae pv. actinidiae is the causative agent of bacterial canker of kiwifruit. The population of this pathogen is differentiated into three biovars, biovar 1, 2 and 3, according to their molecular characteristics. In this work, we determined biovars of P. syringae pv. actinidiae strains isolated in Korea since 1997 and stored in Department of Biology, Sunchon National University, Suncheon, Korea. The biovars of P. syringae pv. actinidiae strains were determined by PCR using biovar specific primers developed previously. Of 682 strains investigated, 288 strains belonged to biovar 2, while 394 strains were biovar 3. There were no P. syringae pv. actinidiae strains belonging to biovar 1 among the strains isolated in Korea. Sudden outbreak and spreading of bacterial canker caused by biovar 3 strain suggest that this strain has character of rapid transmission.

• The Plant Pathology Journal
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• v.31 no.4
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• pp.343-349
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• 2015

Citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is one of the most important bacterial diseases of citrus. Because citrus canker is not found in many countries including European Union and Australia, Xcc is strictly regulated in order to prevent its spread. In this study, the effects of X-irradiation on Xcc growth either in the suspension or on the surface of citrus fruits were investigated. The suspension containing $1{\times}10^7cfu/ml$ of Xcc was irradiated with different absorbed doses of X-irradiation ranging from 50 to 400 Gy. The results showed that Xcc was fully dead at 400 Gy of X-irradiation. To determine the effect of X-irradiation on quarantine, the Xcc-inoculated citrus fruits were irradiated with different X-ray doses at which Xcc was completely inhibited by an irradiation dose of 250 Gy. The $D_{10}$ value for Xcc on citrus fruits was found to be 97 Gy, indicating the possibility of direct application on citrus quarantine without any side sterilizer. Beside, presence of Xcc on the surface of asymptomatic citrus fruits obtained from citrus canker-infected orchards was noted. It indicated that the exporting citrus fruits need any treatment so that Xcc on the citrus fruits should be completely eliminated. Based on these results, ionizing radiation can be considered as an alternative method of eradicating Xcc for export of citrus fruits.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1999.04a
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• pp.26.1-26
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• 1999

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.120.3-121
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• 2003

Relative degree of resistance of citrus to Xanthomonas axonopodis pv. citri, the causal bacterium of canker, was investigated. Growth rate of a bacterium in leaf tissues after infiltration, disease incidence, and percent of lesion area were compared. By using growth rate[(GR=(At - A$\sub$t-1/)/A$\sub$t-1] host plants were differentiated into susceptible and resistant. Growth rates reached to peak at 40 hrs after inoculation and then declined. The growth rate in leaf tissues of a moderately susceptible cultivar, Citrus sinensis vu. Lane late(sweet orange), was the highest, and those of C. unshiu ${\times}$ C. sinensis(kiyomi), C. junos(yuzu), [(Citrus. unshiu x C. sinensis) x C. reticulata] (shiranuhi), and C. unshiu(satuma mandarin) were similar. This result indicates that the growth rate of the bacterium in leaf tissues can be effectively used for evaluation of disease resistance for citrus plants to X. axonopodis pv. citri. The disease on sweet orange occurred earlier than relatively resistant citrus plants tested. The percent of lesion area on leaf was also higher in sweet orange than those of satsuma mandarin, shiranuhi and kiyomi, and yuzu. The disease severity was highest on sweet orange and followed by kiyomi, shiranuhi, satsuma mandarin, and yuzu.