Korean Journal of Agricultural Science (Korean J. Agric. Sci.; KJOAS)
Indexed in KCI (Korea Citation Index), Open Access, Peer Reviewed.
pISSN 2466-2402
eISSN 2466-2410

Identification of virulence-associated genes of Erwinia amylovora by transposon mutagenesis



SY Lee, Kong HG, Kang IJ, Oh H, Woo HJ, Roh E. Identification of virulence-associated genes of Erwinia amylovora by transposon mutagenesis. Korean Journal of Agricultural Science 50:283-289.

Korean Journal of Agricultural Science (Korean J. Agric. Sci.) 2023 June, Volume 50, Issue 2, pages 50:283-289. https://doi.org/10.7744/kjoas.202300019

Received on 31 January 2023, Revised on 27 March 2023, Accepted on 26 April 2023, Published on 30 June 2023.

Identification of virulence-associated genes of Erwinia amylovora by transposon mutagenesis

Seung Yeup Lee1, Hyun Gi Kong2, In Jeong Kang3, Hyeonseok Oh4, Hee-Jong Woo4, Eunjung Roh1,*

1Crop Protection Division, National Institute of Agricultural Sciences, Rural Development Administration, Wanju 55365, Korea

2College of Agriculture, Life and Environment Sciences, Chungbuk National University, Cheongju 28644, Korea

3Department of Central Area Crop Science, National Institute of Crop Science, Suwon 16613, Korea

4National Agrobiodiversity Center, National Institute of Agricultural Sciences, Rural Development Administration, Wanju 55365, Korea

*Corresponding author: rosalia51@korea.kr


Erwinia amylovora, which causes fire blight disease on apple and pear trees, is one of the most important phytopathogens because of its devastating impact. Currently, the only way to effectively control fire blight disease is through the use of antibiotics such as streptomycin, kasugamycin, or oxytetracycline. However, problems with the occurrence of resistant strains due to the overuse of antibiotics are constantly being raised. It is therefore necessary to develop novel disease control methods through an advanced understanding of the pathogenesis mechanism of E. amylovora. To better understand the pathogenesis of E. amylovora, we investigated unknown virulence factors by random mutagenesis and screening. Random mutants were generated by Tn5 transposon insertion, and the pathogenicity of the mutants was assessed by inoculation of the mutants on apple fruitlets. A total of 17 avirulent mutants were found through screening of 960 random mutants. Among them, 14 mutants were already reported as non-pathogenic strains, while three mutants, TS3128_M2899 (ΔSUFU), TS3128_M2939 (ΔwcaG), and TS3128_M3747 (ΔrecB), were not reported. Further study of the association between E. amylovora pathogenicity and these 3 novel genes may provide new insight into the development of control methods for fire blight disease.


Erwinia amylovora, fire blight, transposon mutagenesis, virulence factor


Bayot RG, Ries SM. 1986. Role of motility in apple blossom infection by Erwinia amylovora and studies of fire blight control with attractant and repellent compound. Phytopathology 76:L441-445.

Borruso L, Salomone-Stagni M, Polsinelli I, Schmitt AO, Benini S. 2017. Conservation of Erwinia amylovora pathogenicity-relevant genes among Erwinia genomes. Archives of Microbiology 199:1335-1344.

Bugert P, Geider K. 1995. Molecular analysis of the ams operon required for exopolysaccharide synthesis of Erwinia amylovora. Molecular Microbiology 15:917-933.

Cano DA, Pucciarelli MG, García-del Portillo F, Casadesús J. 2002. Role of the RecBD recombination pathway in Salmonella virulence. Journal of Bacteriology 184:592-595.

Islam R, Brown S, Taheri A, Dumenyo CK. 2019. The gene encoding NAD-dependent epimerase/dehydrase, wcaG, affect cell surface properties, virulence, and extracellular enzyme production in the soft rot phytopathogen, Pectobacterium carotovorum. Microorganisms 7:172.

Lee MS, Lee IG, Kim SK, Oh CS, Park DH. 2018. In vitro screening of antibacterial agents for suppression of fire blight disease in Korea. Plant Disease 24:41-51.

Mun D, Lee J, Choe J, Kim B, Oh S, Song M. 2017. Value of clay as a supplement to swine diets. Korean Journal of Agricultural Science 44:181-187.

Oh CS, Kim JF, Beer SV. 2005. The Hrp pathogenicity island of Erwinia amylovora and identification of three novel genes required for systemic infection. Molecular Plant Pathology 6:125-138.

Piqué N, Miñana-Galbis D, Merino S, Tomàs JM. 2015. Virulence factors of Erwinia amylovora: A review. International Journal of Molecular Sciences 16:12836-12854.

Singer T, Burke E. 2003. High-throughput TAIL-PCR as a tool to identify DNA flanking insertions. Methods in molecular biology, vol 236. Plant functional genomics: Methods and protocols edited by Grotewold E. pp. 241-272. Humana Press, Totowa, NJ, USA.

Slack SM, Walters KJ, Outwater CA, Sundin GW. 2021. Effect of kasugamycin, oxytetracycline, and streptomycin on in-orchard population dynamics of Erwinia amylovora on apple flower stigmas. Plant Disease 105:1843-1850.

Zou GS, Yuan L, Guo W, Li YR, Che YZ, Zou LF, Chen GY. 2010. Construction of a Tn5-tagged mutant library of Xanthomonas oryzae pv. oryzicola as an invaluable resource for functional genomics. Current Microbiology 62:908-916.

Authors Information

Seung Yeup Lee, https://orcid.org/0000-0002-8768-7276

Hyun Gi Kong, Department of plant medicine, Chungbuk National University, Professor

In Jeong Kang, https://orcid.org/0000-0001-6689-0024

Hyeonseok Oh, https://orcid.org/0000-0002-1673-5170

Hee-Jong Woo, https://orcid.org/0000-0003-0922-410X

Eunjung Roh, https://orcid.org/0000-0003-2999-5817

Conflicts of interest

No potential conflict of interest relevant to this article was reported.


본 논문은 농촌진흥청 연구사업(화상병균의 유전체 분석 및 병원성 관련 유전자 선발, PJ01493403)의 지원에 의해 이루어진 것임.