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

• The Plant Pathology Journal
• /
• v.37 no.6
• /
• pp.652-661
• /
• 2021

Xanthomonas citri pv. glycines (Xcg) is a major pathogen of soybean (Glycine max) in South Korea, despite the availability of soybean varieties with some resistance. We conducted a nationwide survey of the incidence and severity of bacterial pustule caused by Xcg. The percentage of infected fields was 7% to 17% between 2015 and 2017. We characterized the diversity of a nationwide collection of 106 Xcg isolates based on avrBs3 banding patterns. The isolates fell into 11 groups, each represented by a type strain; only two of these were similar to isolates collected from 1999 to 2002. The diversity of Xcg strains increased and the dominant strains changed between 1999 and 2017, with three new type strains comprising 44% of the isolates examined in 2012 to 2017. Pathogenicity tests did not show evidence for a shift in the races or aggressiveness of Xcg strains. Korean soybean cultivars, including the widely-grown Daewon cultivar, were susceptible to the 11 new type strains. The cultivar CNS, which carries the rxp resistance gene, was susceptible to most type strains, including two representing 83% of the Korean Xcg strains. In contrast, Williams 82, which also carries rxp, showed resistance to at least five type strains. Collectively, these results suggest that Williams 82 has resistance loci in addition to rxp. The widespread distribution of Xcg, the high virulence of the current endemic strains, and the low resistance of most Korean soybean cultivars collectively favor widespread disease in Korea in years that are favorable to pustule development.

• The Plant Pathology Journal
• /
• v.28 no.2
• /
• pp.207-211
• /
• 2012

It has been known that streptomycin resistance in bacteria can occur as a results of chromosomal mutation or through gene acquisition or both. Chromosomal mutations for resistances are point mutations in the rpsL gene, which alter ribosomal protein S12. Acquired resistance has occurred when an $Sm^R$ plasmid carrying transposon Tn5393 with tandem strA-strB gene is transferred by conjugation. A total of 686 isolates of Xanthomonas smithii subsp. citri causal agent of citrus canker disease were collected from 26 citrus orchards in Jeju Island in 2003 and 2004 seasons. Forty-nine of 111 isolates from streptomycin non-sprayed orchards in 2003 season were resistant to streptomycin. Of 107 isolates from orchards sprayed one time with streptomycin, 58 isolates were resistant, and 166 of 221 isolates from orchards sprayed two times with streptomycin were resistant. In 12 orchards sprayed three or more times with streptomycin, 219 of 247 isolates were resistant to streptomycin. Twenty-five isolates of X. smithii subsp. citri were surveyed to identify the mechanisms of streptomycin resistance in this study. Twenty-one of these 25 isolates were resistant to streptomycin, and it was proven by PCR assay that 18 of the 21 streptomycin resistant isolates have the strB gene. In sixteen of the 21 streptomycin resistant isolates, it was occurred a point mutation altered codon lysine (AAG)-41 of rpsL gene to arginine (AGG). The streptomycin-sensitive isolates easily acquired the resistance by mixed culture with resistant isolates. The strB gene was amplified from the isolates that acquired the resistance by mixed culture, and one isolate of them was also point-mutated in codon 41 of rpsL gene to be resistant. In this study, most of the streptomycin-resistant isolates of X. smithii sub sp. citri in Jeju island expressed the resistance by both chromosomal point mutation and gene acquisition, and the resistance was easily acquired through conjugation by culture mixed with streptomycin resistant and sensitive strains.

• Research in Plant Disease
• /
• v.20 no.2
• /
• pp.101-106
• /
• 2014

Citrus canker caused by Xanthomonas citri subsp. citri (Xcc) is one of the most important diseases on citrus. Although Satsuma mandarin cultivating mostly in Korea is moderately resistance to canker, occurrence of the disease were more frequently reported since last decade. Like other diseases in citrus, citrus canker was mainly protected by chemical fungicide in the field. Due to the side effect of the chemicals, alternative method of disease control is recently required. In this study four rhizobacterial strains TRH423-3, MRL408-3, THJ609-3 and TRH415-2 are selected by testing its antifungal activity against Xcc. Pre-inoculation with the selected rhizobacterial strains caused disease suppression on the citrus leaves after inoculation with the citrus canker pathogen. Similarly, in the field test symptoms of citrus canker were less developed in the citrus trees applied several times with the selected rhizobacterial strains compared with those of untreated trees. Therefore, it is suggested that the selected rhizobacterial strains may be valuable as an alternative method in the environment-friendly citrus farm.

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

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

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2003.10a
• /
• pp.71.1-71
• /
• 2003

Most major plant pathogenic bacteria in Korea belong to Xanthomonas spp.. Xanthomonas oryzae pv. oryzae is a major pathogen in rice, X. campestris pv. vesicatoria in pepper, X. axonopodis pv. giycines in soybean, X. campestris pv. campestris in cabbage, and X. axonoposid pv. citri in tangerin. Host specificity of the bacterial pathogen depends on the avirulence gene in the pathogen and the corresponding resistance gene in host plants. Many avirulence genes in bacteiral pathogen located on the native plasmids. However, the presence of the native plasmids in Xanthomonas spp. was not investigated well. In order to study the host specificity, we isolated native plasmids from Xanthomonas spp. and compared those plasmids each other, The presence of the native plasmids and the characteristics of the plasmids depended on the bacterial strains. In the X. axonopodis pv. glycines, most strains carried native plasmids but some strains did not. Some strains carry about 60 kb native plasmids including 3 different aviurlence genes. We will discuss the characteristics of the native plasmids isolated from the Xanthomonas spp.

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

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

• Microbiology and Biotechnology Letters
• /
• v.32 no.3
• /
• pp.256-264
• /
• 2004

A new primer set was developed for the detection and identification of Xanthomonas oryzae pv. oryzae, the bacterial leaf blight (BLB) pathogen in rice plant. The nucleotide sequence of hpaA gene was determined from X. o. pv. oryzae str. KACC10331, and the sequence information was used to design primers for the application of the polymerase chain reaction (PCR). The nucleotide sequence of hpaA from X. o. pv. oryzae str. KACC 10331 was aligned with those of X. campestris pv. vesicatoria, X. campestris pv. campestris, X. axonopodis pv. citri, and X. axonopodis pv. glycines. Based on these results, a primer set(XOF and XOR) was designed for the specific detection of hpaA in X. o. pv. oryzae. The length of PCR products amplified using the primer set was 534-bp. The PCR product was detected from only X. o. pv. oryzae among other Xanthomonas strains and reference bacteria. This product was used to confirm the conservation of hpaA among Xanthomonas strains by Southern-blotting. Furthermore, PCR amplification with XOF and XOR was used to detect the pathogen in an artificially infected leaf. The sensitivity of PCR detection in the pure culture suspension was also determined. This PCR-based detection methods will be a useful method for the detection and identification of X. o. pv. oryzae as well as disease forecasting.