• 제목/요약/키워드: Citrus canker

검색결과 25건 처리시간 0.023초

Difference of Physiochemical Characteristics Between Citrus Bacterial Canker Pathotypes and Identification of Korean Isolates with Repetitive Sequence PCRs

  • Lee, Yong-Hoon;Lee, Seung-Don;Lee, Dong-Hee;Yu, Sang-Mi;Lee, Jung-Hee;Heu, Sung-Gi;Hyun, Jae-Wook;Ra, Dong-Soo;Park, Eun-Woo
    • The Plant Pathology Journal
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    • 제24권4호
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    • pp.423-432
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    • 2008
  • The difference of carbon source utilization and fatty acid composition between the pathotypes of Xanthomonas strains, which causing citrus bacterial canker was compared, and the physiochemical characteristics were used to analyze relationship of the strains for the first time. The pattern of several carbon sources utilization and fatty acids composition reliably discriminated the pathotypes of Xanthomonas strains. The dendrogram which was constructed by 95 carbon source utilization profiles differentiated X. axonopodis pv. citri A, $A^*$ and $A^w$ from the other pathotypes. When the dendrogram was drawn by combined analysis of carbon source utilization pattern and fatty acid composition, X. axonopodis pv. aurantifolii B, C and X. axonopodis pv. citrumelo formed a distinct cluster. The difference of carbon source utilization and fatty acid composition could be used effectively for the identification of pathotypes of citrus bacterial canker. The physiochemical characteristics strongly indicated that the strains isolated in Korea belong to X. axonopodis pv. citri A type. The cluster analysis by the band patterns of ERIC-, BOX- and REP-PCR allowed the discrimination of the pathotypes isolated from Korea. However, the rep-PCRs could not differentiate X. axonopodis pv. citri A types from $A^*$ and $A^w$ types. The overall results of metabolic profiles and rep-PCRs strongly indicated that the Korean isolates are X. axonopodis pv. citri A type.

Mode of Action of Streptomycin Resistance in the Citrus Canker Pathogen (Xanthomonas smithii subsp. citri) in Jeju Island

  • Hyun, Jae-Wook;Kim, Hyo-Jung;Yi, Pyoung-Ho;Hwang, Rok-Yeon;Park, Eun-Woo
    • The Plant Pathology Journal
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    • 제28권2호
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    • pp.207-211
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    • 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.

Characterization of the host reaction of some citrus plants with Xanthomonas axonopodis pv. citri, causing citrus bacterial canker disease.

  • Myung, Inn-Shik;Hyun, Jae-Wook;Kim, Kwang-Sik;Lee, Sung-Chan;Lim, Han-Chul
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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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.

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Dispersal of Xanthomonas axonopodis pv. citri, the Causal Bacterium of Citrus Canker, on Unshiu Orange.

  • Myung, Inn-Shik;Nam, Ki-Woong;Kwon, Hyeog-Mo
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.121.1-121
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    • 2003
  • Dispersal of Xanthomonas axonopodis pv. citri, causing citrus bacterial canker disease on Unshiu orange was investigated at previously infested plots at Seogwipo in Jeju island of Korea. The bacterial pathogen overwintered in lesions started to multiply at tate May, and disease firstly observed one month after detection of phage from lesions. The disease gradually increased, however, it dispersed non-directionally to nearby plants from inoculum sources. Diseased plants were aggregated to form a cluster throughout the experiment. Population dynamics of phage on symtomless leaf surface and the disease severity were compared in the nursery, Increase of phage population on symptomless leaf surface preceded one month to that of the disease severity Population of phage increased constantly from late July to October, however, the disease severity decreased from late August to late October. It was assumed that the decrease of disease severity might be due to disease-induced defoliation.

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Studies on Physiology, Ecology and Protection of Citrus Canker Caused by Xanthomonas axonopodis pv, citri

  • Lee, Seong-Chan;Hyun, Jae-Wook;Kim, Dong-Hwan;Kim, Kwang-Sik;Lim, Han-Chul
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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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.

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Different Profiles of the Negatively Stained Citrus Canker Bacterium Xanthomonas citri pv. citri Depending on Culture Media and Heavy Metal Stains

  • Kim, Ki-Woo;Lee, In-Jung;Hyun, Jae-Wook;Lee, Yong-Hoon;Park, Eun-Woo
    • The Plant Pathology Journal
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    • 제26권1호
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    • pp.90-92
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    • 2010
  • Staining profiles and bacterial morphology were compared in Xanthomonas citri pv. citri by a transmission electron microscopy. Four types of negative staining regimes were employed depending on culture media and heavy metal stains. The bacterial cells grown on LB agar media often appeared clustered on the supporting film. Meanwhile, individual bacterial cells could be readily found on the preparations from LB broth media. Typical rod-shaped cells (ca. $1\;{\mu}m$ in length) and their flagella were observed in either 2% uranyl acetate (UA) or 2% neutralized potassium phosphotungstate (PTA) staining. The UA-stained bacteria often showed relatively intact cell morphology and rather positively stained cells with a thin electron-dense stain depth around bacteria. The PTA-stained bacteria were characterized by the wrinkled cell surface where the stain was entrapped in grooves. In addition, distinct electron-dense stain depth was evident around the PTA-stained preparations. Numerous fimbriae could be mostly observed from the PTA-stained preparations of the two culture media, but not from the UA-stained preparations.

최근 주요 감귤 병의 발생 양상 (Aspect of Incidence of the Major Citrus Diseases Recently)

  • 현재욱;이평호;황록연;문경환
    • 식물병연구
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    • 제19권2호
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    • pp.102-107
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    • 2013
  • 2003년도부터 감귤검은점무늬병에 걸린 과실율을 조사해 본 결과 2010년에 16.8%로써 가장 심하였으며 2005년에는 3.8%로써 가장 낮았고 10년간 평균 병든 과율은 8.8%이었다. 지역별로 최근 10년 동안 병든 과율을 비교해 보면 제주도 남동부, 동부지역에서 병 발생이 많았다. 또한 발병도를 조사해 본 결과 2003, 2004, 2007, 2010년 검은점무늬병 발병도가 각각 17.0, 22.6, 19.2, 18.9%로써 다른 해에 비하여 발병도가 높았다. 궤양병의 경우 2004년에 2.19% 발병하여 최근 들어 가장 많이 발생하였으며 그 이후로 급격히 줄어들어 2009년과 2010년의 경우 0.08와 0.10%이었지만, 2011년의 경우 1.50%로써 다시 증가하였으며 2004년부터 2012년까지 평균 발생율은 0.81%이었다. 또한 발생 지역도 제주도 동부와 서부지역에서 많았다. 더뎅이병의 경우 2004년부터 2012년까지 9년간 평균 발생과율은 0.25% 이었으며 2004년 0.72%로 가장 발생이 많았다. 감귤 과실에서의 검은점무늬병 진전 상황을 조사해 본 결과 해에 따라 약간씩 상이하지만 7월 하순부터 8월 하순까지 병 진전이 가장 많았다. 더뎅이병의 경우 5월 10일경에 첫 병징이 나타났으며 이후 5월 하순에서 6월 중순사이에 급격히 증가하였다. 궤양병의 경우 6월 중순경에 첫 병징이 나타나서 6월 하순까지 봄순에서 병이 진전되었다. 죽은 가지에서 검은점무늬병 전염원 주 발생 시기는 6월 하순부터 8월 상순경이었다.

Evaluation of Potential of Mandarin Hybrid 'Shiranuhi' against inoculation of Bacterial Canker Disease Pathogen (Xanthomonas axonopodis pv. citri) in Citrus Field in Jeju Island

  • Hyun, Jae-Wook;Myung, Inn-Shik;Lee, Seong-Chan;Kim, Kwang-Sik;Lim, Han-Cheol
    • The Plant Pathology Journal
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    • 제19권5호
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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.

감마선에 의한 감귤 궤양병균의 불활성화와 감귤 궤양병 감염에 미치는 영향 (Inactivation of Xanthomonas citri subsp. citri and Effect on Infection of Citrus Canker by Gamma Irradiation)

  • 김경남;송민아;한상헌;송성준;전용철
    • 식물병연구
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    • 제20권4호
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    • pp.283-288
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    • 2014
  • 감귤 궤양병은 Xanthomonas citri subsp. citri에 의해 발생하는 병으로서 많은 나라에서 이 병에 대해 검역을 실시하고 있다. 최근 methyl bromide의 검역 시 사용이 제한됨에 따라 감마선을 이용한 농수축산물에 대한 살균 방안이 대두되고 있다. 본 연구에서는 현탁액 형태의 감귤 궤양병균과 생과실 과피에 존재하는 감귤 궤양병균의 밀도를 감소시킬 수 있는 감마선의 선량을 알아보았다. 미생물의 밀도를 90% 감소시키는 선량인 감귤 궤양병균의 $D_{10}$ value는 현탁액과 감귤 생과실 과피에서 각각 55와 28 Gy였다. 또한 감마선을 처리한 감귤 궤양병균 현탁액을 감귤 잎에 접종하였더니 병 발생이 억제되었다. 본 연구를 통하여 감마선 처리에 의해 감귤 생과실에 존재하는 감귤 궤양병균을 박멸할 수 있다고 판단되며 본 연구 결과는 감마선을 검역에 활용하는데 가치가 있다고 생각된다.

Detection of Xanthomonas axonopodis pv. citri on Citrus Fruits Using Enzyme-Linked Immunosorbent Assay

  • Jin, Kyoung-Sik;Kang, Ik-Beom;Ko, Kyoung-Il;Lee, Eun-Seob;Heo, Jong-Young;Kang, Young-Kil;Kim, Byung-Ki
    • The Plant Pathology Journal
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    • 제17권1호
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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.

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