• 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.

• The Plant Pathology Journal
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• v.17 no.1
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• pp.62-66
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• 2001

Detection of Xanthomonas axonopodis pv. citri (Xac) on citrus fruits for exporting is usually made by bacteriophage test (BPT) to demonstrate the pathogen-free status. BPT has rather time-consuming and complicate procedures for dealing with massive samples to be inspected. In this study, enzyme-linked immunosorbent assay (ELISA) was applied to detect Xac on fruits, and compared with BPT. In ELISA, positive reactions occurred in the bacterial densities of $3\times10^5$ cells/ml or more. To detect the bacterial infection on citrus fruits with a density of lower than $3\times10^5$ cells/ml, the bacterial suspensions were mixed with fruit rinse water and incubated in broth medium. Ordinary peptone sucrose broth (PSB) was not a proper medium for increasing Xac density specifically enough to be detect by ELISA. On the other hand, modified PSB (MPSP) amended with Fe-EDTA (0.25 g/$\ell$) and 2.5% potato-dextrose broth sufficed to differentiate uninfected and infected citrus fruits by ELISA after 24 h incubation of the fruit rinse water. Using various citrus samples from infected and uninfected fields, efficiencies in detecting Xac on fruits were compared between ELISA and BPT. For infected fruits samples, ELISA detected Xac by 100%, while BPT by about 44%, indicating that the detection efficiency was improved by 23.5% by ELISA, compared to BPT. In addition, ELISA has simpler procedures for testing and is less time-consuming than BPT, suggesting that ELISA may be accurate and simple method to detect Xac on citrus fruits.

• 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.

• The Plant Pathology Journal
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• v.19 no.5
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• pp.248-252
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• 2003

This study was carried out to evaluate potential of Shiranuhi cultivar against inoculation of causal pathogen of citrus canker, Xanthomonas axonopodis pv. citri in Jeju Island by comparing degree of susceptibility of fruits and leaves/twigs, and analysis of incidence of canker disease. Progression of symptom, disease incidence, and percent area of lesion were surveyed for evaluation of resistance. In greenhouse condition, symptoms of bacterial citrus canker progressed more rapidly in sweet orange, a moderately susceptible cultivar, than in other four cultivars (satsuma mandarin, 'Kiyomi', 'Shiranuhi' and 'Yuzu'). At 20 days after inoculation, disease severity was the highest in sweet orange (5.0$\pm$0.0), and all tested leaves were distorted or had dropped. On the other hand, 'Yuzu' showed the lowest disease severity (2.6$\pm$0.47), followed by 'Kiyomi' (4.0$\pm$0.0), 'Shiranuhi' (4.0$\pm$0.82), and satsuma mandarin (4.3$\pm$0.47). Percent area of lesion per leaf 30 days after inoculation was the highest in sweet orange (8.31$\pm$1.78), followed by satsuma mandarin (1.51$\pm$1.25), 'Shiranuhi' (1.39$\pm$0.94), and 'Kiyomi' (1.1$\pm$0.9), while the lowest was in 'Yuzu' (0.26$\pm$0.17). Infield condition, percentage of diseased leaf in 'Shiranuhi' was very low, 5.2$\pm$2.9, compared with sweet orange, 71.0$\pm$ 11.5, while that of satsuma mandarin and 'Kiyomi' were 6.9$\pm$7.0 and 4.3$\pm$2.0, respectively. Percentages of diseased leaf was higher (17.4$\pm$7.1) than that of diseased fruit (3.2$\pm$2.5) in severely diseased trees of Shiranuhi cultivar, and the disease was not observed on twig in open field condition. Lesion sizes on leaves and fruits in open field condition were 4.1$\pm$2.2 mm2 and 5.1$\pm$5.6 mm2, respectively, while those in greenhouse condition were 8.7$\pm$5.7 mm2, 10.4$\pm$9.2 mm2 and 5.6$\pm$2.6 mm2 on leaves, fruits and twigs, respectively. The disease was observed in 5.6% out of total 107 farmers Shiranuhi fields under polyethylene film house, and average percentages of diseased tree in 31 fields of Shiranuhi cultivar and adjacent satsuma mandarin fields were 0.02% and 14.8%, respectively. Average percentage of diseased fruit was 1.6% in satsuma mandarin which was not observed in anyone of all the 31 Shiranuhi farmers fields. Therefore, it was concluded that 'Shiranuhi' cultivar is not potential against causal pathogen of citrus canker disease in Jeju Island because the cultivar has similar resistance as satsuma mandarin which occupies over 95% of total 25,000 ha in Jeju Island in polyethylene film houses protected from outside.

• The Korean Journal of Pesticide Science
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• v.13 no.1
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• pp.45-53
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• 2009

An antagonistic bacterial strain KLF01 was isolated from rhizosphere of tomato and identified to be Lactobacillus sp. by biochemical and genetic analysis. This strain showed antagonism against the used plant pathogenic bacteria like Ralstonia solanacearum, (bacterial wilt), Xanthomonas axonopodis pv. citri, (Citrus canker), Xanthomonas campestris pv. vesicatoria (Bacterial spot), Eriwinia pyrifoliae (Shoot-blight) and Eriwinia carotovora subsp. carotovora group (Potato scab) through agar well diffusion method. In planta test done by drench application of strain KLF01 $(4{\times}10^8 cfu/ml)$ into the experimental plot containing tomato (Solanum lycopersicum L.) cultivar 'Lokkusanmaru' and red pepper (Capsicum annuum L.) cultivar 'Buja' plants, in pot test post-inoculated with the plant pathogenic bacteria, R. solanacearum significantly reduced the disease severity, compared to the non-treated plants.

• The Plant Pathology Journal
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• v.18 no.6
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• pp.333-337
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• 2002

Bacteriophage of Xanthomonas axonopodis pv. citri, a causal agent of citrus canker disease, was studied for its cultural characteristics. The relative efficiency of plat-ing (EOP) of 11 phages used to 13 strains off, axonopodis pv. citri tested ranged from 0.8 to 1, indicating that the phages are homogeneous. Homogeneity of the phages suggests that citrusphage belongs to a single group CPK as reported in a previous study. Typical one-step growth of a phage P5 selected from the citrusphages was observed. The EOP of the P5 was dependent upon the media, pH, and temperature. It was observed that multiplication of the phage cultured in Wakimotos potato semisynthetic media at $25^{\circ}C$ was more effective than that in other temperatures, regardless of the bacterial strains and media used. It was observed that pH 6.5 is optimal for multiplication of the phage. In comparison of the EOP among citrusphages $CP_1$, $CP_2$, and P5, multiplicative characteristic of phage P5 in the bacteria on time-course was similar with that of phage $CP_1$. Thus, it was concluded that citrusphage group CPK from Korea is $CP_1$ based on host specificity of the phage as described in a previous study, homogeneity, and its multiplication pattern.