Browse > Article
http://dx.doi.org/10.5423/PPJ.OA.03.2022.0027

Discrimination and Detection of Erwinia amylovora and Erwinia pyrifoliae with a Single Primer Set  

Ham, Hyeonheui (Crop Protection Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Kim, Kyongnim (Crop Protection Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Yang, Suin (Crop Protection Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Kong, Hyun Gi (Crop Protection Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Lee, Mi-Hyun (Crop Protection Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Jin, Yong Ju (Crop Protection Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Park, Dong Suk (Crop Protection Division, Department of Agro-food Safety and Crop Protection, National Institute of Agricultural Sciences, Rural Development Administration)
Publication Information
The Plant Pathology Journal / v.38, no.3, 2022 , pp. 194-202 More about this Journal
Abstract
Erwinia amylovora and Erwinia pyrifoliae cause fire blight and black-shoot blight, respectively, in apples and pears. E. pyrifoliae is less pathogenic and has a narrower host range than that of E. amylovora. Fire blight and black-shoot blight exhibit similar symptoms, making it difficult to distinguish one bacterial disease from the other. Molecular tools that differentiate fire blight from black-shoot blight could guide in the implementation of appropriate management strategies to control both diseases. In this study, a primer set was developed to detect and distinguish E. amylovora from E. pyrifoliae by conventional polymerase chain reaction (PCR). The primers produced amplicons of different sizes that were specific to each bacterial species. PCR products from E. amylovora and E. pyrifoliae cells at concentrations of 104 cfu/ml and 107 cfu/ml, respectively, were amplified, which demonstrated sufficient primer detection sensitivity. This primer set provides a simple molecular tool to distinguish between two types of bacterial diseases with similar symptoms.
Keywords
detection; discrimination; Erwinia amylovora; Erwinia pyrifoliae; polymerase chain reaction;
Citations & Related Records
Times Cited By KSCI : 3  (Citation Analysis)
연도 인용수 순위
1 Ye, J., Coulouris, G., Zaretskaya, I., Cutcutache, I., Rozen, S. and Madden, T. L. 2012. Primer-BLAST: a tool to design targetspecific primers for polymerase chain reaction. BMC Bioinform. 13:134.   DOI
2 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.   DOI
3 Samson, R., Legendre, J. B., Christen, R., Saux, M. F.-L., Achouak, W. and Gardan, L. 2005. Transfer of Pectobacterium chrysanthemi (Burkholder et al., 1953) Brenner et al. 1973 and Brenneria paradisiaca to the genus Dickeya gen. nov. as Dickeya chrysanthemi comb. nov. and Dickeya paradisiaca comb. nov. and delineation of four novel species, Dickeya dadantii sp. nov., Dickeya dianthicola sp. nov., Dickeya dieffenbachiae sp. nov. and Dickeya zeae sp. nov. Int. J. Syst. Evol. Microbiol. 55:1415-1427.   DOI
4 Schuler, G. D. 1997. Sequence mapping by electronic PCR. Genome Res. 7:541-550.   DOI
5 Taylor, R. K., Guilford, P. J., Clark, R. G., Hale, C. N. and Forster, R. L. S. 2001. Detection of Erwinia amylovora in plant material using novel polymerase chain reaction (PCR) primers. N. Z. J. Crop Hortic. Sci. 29:35-43.   DOI
6 Banerji, S. and Flieger, A. 2004. Patatin-like proteins: a new family of lipolytic enzymes present in bacteria? Microbiology 150:522-525.   DOI
7 Bonn, W. G. and van der Zwet, T. 2000. Distribution and economic importance of fire blight. In: Fire blight: the disease and its causative agent, Erwinia amylovora, ed. by J. L. Vanneste, pp. 37-53. CABI Publishing, Wallingford, UK.
8 Bereswill, S., Bugert, P., Bruchmuller, I. and Geider, K. 1995. Identification of the fire blight pathogen, Erwinia amylovora, by PCR assays with chromosomal DNA. Appl. Environ. Microbiol. 61:2636-2642.   DOI
9 Bereswill, S., Pahl, A., Bellemann, P., Zeller, W. and Geider, K. 1992. Sensitive and species-specific detection of Erwinia amylovora by polymerase chain reaction analysis. Appl. Environ. Microbiol. 58:3522-3526.   DOI
10 Billing, E. 2011. Fire blight. Why do views on host invasion by Erwinia amylovora differ? Plant Pathol. 60:178-189.   DOI
11 Denning, W. 1794. On the decay of apple trees. Trans. Soc. Promotion Agric. Arts Manufact. Inst. State N. Y. 2:219-222.
12 Drenova, N. V., Isin, M. M., Dzhaimurzina, A. A., Zharmukhamedova, G. A. and Aitkulov, A. K. 2012. Bacterial fire blight in the Republic of Kazakhstan. Plant Health Res. Pract. 1:44-48.
13 Fatmi, M., Bougsiba, M. and Saoud, H. 2008. First report of fire blight caused by Erwinia amylovora on pear, apple, and quince in Morocco. Plant Dis. 92:314.
14 Ginzinger, D. G. 2002. Gene quantification using real-time quantitative PCR: an emerging technology hits the mainstream. Exp. Hematol. 30:503-512.   DOI
15 Ham, H., Lee, Y.-K., Kong, H. G., Hong, S. J., Lee, K. J., Oh, G.-R., Lee M.-H. and Lee, Y. H. 2020. Outbreak of fire blight of apple and Asian pear in 2015-2019 in Korea. Res. Plant Dis. 26:222-228 (in Korean).   DOI
16 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 amylovora in Korea. Plant Dis. 100:1946.
17 Jones, A. L. and Geider, K. 2001. Gram negative bacteria, Erwinia amylovora group. In: Laboratory guide for identification of plant pathogenic bacteria, eds, by N. W. Schaad, J. B. Jones and W. Chun, pp. 40-55. APS Press, St. Paul, MN, USA.
18 Kado, C. I. 2006. Erwinia and related genera. In: The prokaryotes. A handbook on the biology bacteria: proteobacteria: gamma subclass, eds. by M. Dworkin, S. Falkow, E. Rosenberg, K.-H. Schleifer, E. Stackebrandt, 3rd ed., pp. 443-450. Springer, New York, NY, USA.
19 Rozen, S. and Skaletsky, H. 2000. Primer3 on the WWW for general users and for biologist programmers. Methods Mol. Biol. 132:365-386.
20 Thomson, S. V. 2000. Epidemiology of fire blight. In: Fire blight: the disease and its causative agent, Erwinia amylovora, ed. by J. L. Vanneste, pp. 9-36. CABI Publishing, Wallingford, UK.
21 Ivanovic, M., Kuzmanovic, N., Gasic, K., Prokic, A., Zlatkovic, N. and Obradovic, A. 2019. Specificity and sensitivity of three PCR-based methods for detection of Erwinia amylovora in pure culture and plant material. Genetika 51:1039-1052.   DOI
22 Lee, G. M., Ko, S., Oh, E.-J., Song, Y.-R., Kim, D. and Oh, C.-S. 2020. Comparative genome analysis reveals natural variations in the genomes of Erwinia pyrifoliae, a black shoot blight pathogen in apple and pear. Plant Pathol. J. 36:428-439.   DOI
23 Sipos, R., Szekely, A. J., Palatinszky, M., Revesz, S., Marialigeti, K. and Nikolausz, M. 2007. Effect of primer mismatch, annealing temperature and PCR cycle number on 16S rRNA gene-targetting bacterial community analysis. FEMS Microbiol. Ecol. 60:341-350.   DOI
24 Wenneker, M. and Bergsma-Vlami, M. 2015. Erwinia pyrifoliae, a new pathogen on strawberry in the Netherlands. J. Berry Res. 5:17-22.   DOI
25 European and Mediterranean Plant Protection Organization. 2013. PM 7/20 (2) Erwinia amylovora. Bull. OEPP/EPPO Bull. 43:21-45.   DOI
26 Llop, P., Donat, V., Rodriguez, M., Cabrefiga, J., Ruz, L., Palomo, J. L., Montesinos, E. and Lopez, M. M. 2006. An indigenous virulent strain of Erwinia amylovora lacking the ubiquitous plasmid pEA29. Phytopathology 96:900-907.   DOI
27 Kralik, P. and Ricchi, M. 2017. A basic guide to real time PCR in microbial diagnostics: definitions, parameters, and everything. Front. Microbiol. 8:108.
28 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. 49:899-906.   DOI
29 Kim, W. S., Jock, S., Paulin, J.-P., Rhim, S.-L. and Geider, K. 2001. Molecular detection and differentiation of Erwinia pyrifoliae and host range analysis of the Asian pear pathogen. Plant Dis. 85:1183-1188.   DOI
30 Lang, J. M., Hamilton, J. P., Diaz, M. G. Q., Van Sluys, M. A., Burgos, M. R. G., Vera Cruz, C. M., Buell, C. R., Tisserat, N. A. and Leach, J. E. 2010. Genomics-based diagnostic marker development for Xanthomonas oryzae pv. oryzae and X. oryzae pv. oryzicola. Plant Dis. 94:311-319.   DOI
31 Mohammadi, M., Moltmann, E., Zeller, W. and Geider, K. 2009. Characterisation of naturally occurring Erwinia amylovora strains lacking the common plasmid pEA29 and their detection with real-time PCR. Eur. J. Plant Pathol. 124:293-302.   DOI
32 Obradovic, D., Balaz, J. and Kevresan, S. 2007. Detection of Erwinia amylovora by novel chromosomal polymerase chain reaction primers. Microbiology 76:748-756.   DOI
33 Park, D. H., Thapa, S. P., Kim, W.-S., Hur, J. H. and Lim, C. K. 2010. Species-specific detection of Erwinia pyrifoliae by PCR assay using enterobacterial repetivive intergenic consensus (ERIC) primers. Plant Pathol. J. 26:267-270.   DOI
34 Powney, R., Beer, S. V., 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.   DOI