Research in Plant Disease (식물병연구)

The Korean Society of Plant Pathology (한국식물병리학회)
권호 표지
DOI QR코드

DOI QR Code

On-Site Diagnosis of Fire Blight with Antibody-Based Diagnostic Strips

항혈청 기반 진단 스트립을 이용한 과수 화상병 현장진단

  • Heo, Gwang-Il (Major in Plant Medicine, School of Applied Plant Science & Biotechnology, Chungbuk National University) ;
  • Shin, Doo-San (Major in Plant Medicine, School of Applied Plant Science & Biotechnology, Chungbuk National University) ;
  • Son, Soo-Hyeong (Major in Plant Medicine, School of Applied Plant Science & Biotechnology, Chungbuk National University) ;
  • Oh, Chang-Sik (Department of Horticultural Biotechnology, Kyung Hee University) ;
  • Park, Duck Hwan (Applied Biology Program, Division of Bioresource Sciences, Kangwon National University) ;
  • Lee, Young-Kee (Department of Agro-food Safety and Crop Protection, National Institute of Agriculture Sciences, Rural Development Administration) ;
  • Cha, Jae-Soon (Major in Plant Medicine, School of Applied Plant Science & Biotechnology, Chungbuk National University)
  • 허광일 (충북대학교 응용생물공학부 식물의학전공) ;
  • 신두산 (충북대학교 응용생물공학부 식물의학전공) ;
  • 손수형 (충북대학교 응용생물공학부 식물의학전공) ;
  • 오창식 (경희대학교 원예생명공학과) ;
  • 박덕환 (강원대학교 생물자원과학부 응용생물학전공) ;
  • 이영기 (농촌진흥청 국립농업과학원 농산물안전성부) ;
  • 차재순 (충북대학교 응용생물공학부 식물의학전공)
  • Received : 2017.07.17
  • Accepted : 2017.08.17
  • Published : 2017.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

Recently fire blight occurred in the Republic of Korea and eradication program for the disease has been executed since then. Specificity and detection sensitivity of the 2 antibody-based diagnostic strips to Korean isolates of Erwinia amylovora (Ea) and their application for on-site diagnosis were evaluated in this study. Ea AgriStrip, a commercial diagnostic kit, and EB strip, developed in this study, reacted positively to the all tested Korean Ea strains and also to most of Erwinia pyrifoliae (Ep) strains causing black shoot blight. They reacted negatively to all Pusedomonas syringae pv. syringae (Pss) strains that cause shoot blight on apple. Detection sensitivity was similar between the 2 strips. For on-site diagnosis, the two strips reacted positively only to the extractions of the fire-blighted samples on all fire blight occurred orchards except one orchard at which on-site diagnosis was carried out at winter time. In addition, they reacted positively to the black-shoot blighted extractions from the black shoot blight occurred apple orchard. These results suggest that both EB strip and Ea AgriStrip would be useful for on-site diagnosis of fire blight in Korea.

최근 한국에서 화상병이 발생하였으며, 그 이후로 이병에 대한 박멸프로그램이 가동되고 있다. 본 연구에서는 2가지 항혈청기반 진단 스트립의 국내 분리 화상병균 Erwinia amylovora (Ea)에 대한 특이성과 검출민감도 그리고 현장진단에 적용을 평가하였다. 상업화된 Ea AgriStrip과 본 연구에서 개발한 EB strip은 검정에 사용한 모든 한국 Ea 균주들과 가지검은마름병을 일으키는 대부분의 Erwinia pyrifoliae (Ep)에 양성반응을 보여주었다. 사과나무 가지마름병을 일으키는 Pusedomonas syringae pv. syringae (Pss)의 사용한 모든 균주에 음성반응을 보여주었다. 2 스트립 사이에 검출민감도는 유사하였다. 현장진단의 경우, 2가지 스트립은 화상병이 발생한 모든 과수원에서 채취한 화상병징 세균추출액에 대하여 양성반응을 보였다. 그러나 겨울철에 진행된 한 곳의 화상병 현장진단에서는 화상병병징 세균추출액에 음성반응을 보였다. 또한 2가지 스트립은 가지검은마름병이 발생한 과수원에서 채집한 가지검은마름병징 세균추출액에 대해서 양성반응을 보여주었다. 이상의 결과는 Ea AgriStrip과 EB strip이 화상병 현장진단에 유용할 것으로 암시한다.

Keywords

References

  1. Braun-Kiewnick, A., Altenbach, D., Oberhansli, T., Bitterlin, W. and Duffy, B. 2011. A rapid lateral-flow immunoassay for phytosanitary detection of Erwinia amylovora and on-site fire blight diagnosis. J. Microbiol. Methods 87: 1-9. https://doi.org/10.1016/j.mimet.2011.06.015
  2. Buhlmann, A., Pothier, J. F., Rezzonico, F., Smits, T. H. M., Andreou, M., Boonham, N., Duffy, B. and Frey, J. E. 2013. Erwinia amylovora loop-mediated isothermal amplification (LAMP) assay for rapid pathogen detection and on-site diagnosis of fire blight. J. Microbiol. Methods 92: 332-339. https://doi.org/10.1016/j.mimet.2012.12.017
  3. EPPO. 2013. PM 7/20 (2) Erwinia amylovora. Bull. OEPP/EPPO Bull. 43: 21-45. https://doi.org/10.1111/epp.12019
  4. Gorris, M. T., Cambra, E., Paulin, J. P., Chartier, R., Cambra, M. and Lopez, M. M. 1996 Production and characterization of monoclonal antibodies specific for Erwinia amylovora and their use in different serological techniques. Acta Hortic. 411: 47-52.
  5. Han, K. S., Yu, J.-G., Lee, H.-B., Oh, C.-S., Yea, M.-C., Lee, J.-H. and Park, D. H. 2016. Controlling by effective pruning of twigs showing black shoot blight disease symptoms in apple trees. Res. Plant Dis. 22: 269-275. https://doi.org/10.5423/RPD.2016.22.4.269
  6. Kim, W. S., Gardan, L., Rhim, S. L. and Geider, K. 1999. Erwinia pyrifoliae sp. nov., a novel pathogen that affects Asian pear trees (Pyrus pyrifolia Nakai). Int. J. Syst. Bacteriol. 49 Pt 2: 899-905. https://doi.org/10.1099/00207713-49-2-899
  7. Myung, I.-S., Lee, J.-Y., Yun, M.-J., Lee, Y.-H., Lee, Y.-K., Park, D. H. and Oh, C.-S. 2016. Fire blight of apple, caused by Erwinia amylovora, a new disease in Korea. Plant Dis. 100: 1774.
  8. Park, D. H., Lee, Y.-G., Kim, J.-S., Cha, J.-S. and Oh, C.-S. 2017. Current status of fire blight caused by Erwinia amylovora and action for its management in Korea. J. Plant Pathol. doi: 10.4454/jpp.v99i0.3918/ (In press)
  9. Park, D. H., Yu, J.-G., Oh, E.-J., Han, K. S., Yea, M. C., Lee, S. J., Myung, I.-S., Shim, H. S. and Oh, C.-S. 2016. First report of fire blight disease on Asian pear caused by Erwinia amylovorain Korea. Plant Dis. 100: 1946.
  10. Powney, R., Beer, S., Plummer, K., Luck, J. and Rodoni, B. 2011. The specificity of PCR-based protocols for detection of Erwinia amylovora. Australas. Plant Pathol. 40: 87-97. https://doi.org/10.1007/s13313-010-0017-7
  11. Rhim, S. L., Volksch, B., Gardan, L., Paulin, J. P., Langlotz, C., Kim, W. S. and Geider, K. 1999. Erwinia pyrifoliae, an Erwinia species different from Erwinia amylovora, causes a necrotic disease of Asian pear trees. Plant Pathol. 48: 514-520. https://doi.org/10.1046/j.1365-3059.1999.00376.x
  12. Shrestha, R., Lee, S. H., Kim, J. E., Wilson, C., Choi, S. G., Park, D. H., Wang, M. H., Hur, J. H. and Lim, C. K. 2007. Diversity and detection of Korean Erwinia pyrifoliae strains as determined by plasmid profiling, phylogenetic analysis and PCR. Plant Pathol. 56: 1023-1031. https://doi.org/10.1111/j.1365-3059.2007.01679.x
  13. The Korean Society of Plant Pathology. 2009. List of Plant Diseases in Korea. 5th ed. Suwon, Korea. 853 pp.
  14. van der Zwet, T., Orolaza-Halbrent, N. and Zeller, W. 2012. Fire Blight, History, Biology, and Management, pp. 15-36. APS Press, St. Paul, MN, USA.