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dc.contributor.authorBorn, Yannick-
dc.contributor.authorFieseler, Lars-
dc.contributor.authorThöny, Valentin-
dc.contributor.authorLeimer, Nadja-
dc.contributor.authorDuffy, Brion-
dc.contributor.authorLoessner, Martin J.-
dc.date.accessioned2018-10-29T13:38:11Z-
dc.date.available2018-10-29T13:38:11Z-
dc.date.issued2017-06-
dc.identifier.issn0099-2240de_CH
dc.identifier.issn1098-5336de_CH
dc.identifier.urihttps://digitalcollection.zhaw.ch/handle/11475/12255-
dc.description.abstractErwinia amylovora is the causative agent of fire blight, a devastating plant disease affecting members of the Rosaceae Alternatives to antibiotics for control of fire blight symptoms and outbreaks are highly desirable, due to increasing drug resistance and tight regulatory restrictions. Moreover, the available diagnostic methods either lack sensitivity, lack speed, or are unable to discriminate between live and dead bacteria. Owing to their extreme biological specificity, bacteriophages are promising alternatives for both aims. In this study, the virulent broad-host-range E. amylovora virus Y2 was engineered to enhance its killing activity and for use as a luciferase reporter phage, respectively. Toward these aims, a depolymerase gene of E. amylovora virus L1 (dpoL1-C) or a bacterial luxAB fusion was introduced into the genome of Y2 by homologous recombination. The genes were placed downstream of the major capsid protein orf68, under the control of the native promoter. The modifications did not affect viability of infectivity of the recombinant viruses. Phage Y2::dpoL1-C demonstrated synergistic activity between the depolymerase degrading the exopolysaccharide capsule and phage infection, which greatly enhanced bacterial killing. It also significantly reduced the ability of E. amylovora to colonize the surface of detached flowers. The reporter phage Y2::luxAB transduced bacterial luciferase into host cells and induced synthesis of large amounts of a LuxAB luciferase fusion. After the addition of aldehyde substrate, bioluminescence could be readily monitored, and this enabled rapid and specific detection of low numbers of viable bacteria, without enrichment, both in vitro and in plant material. IMPORTANCE: Fire blight, caused by Erwinia amylovora, is the major threat to global pome fruit production, with high economic losses every year. Bacteriophages represent promising alternatives to not only control the disease, but also for rapid diagnostics. To enhance biocontrol efficacy, we combined the desired properties of two phages, Y2 (broad host range) and L1 (depolymerase for capsule degradation) in a single recombinant phage. This phage showed enhanced biocontrol and could reduce E. amylovora on flowers. Phage Y2 was also genetically engineered into a luciferase reporter phage, which transduces bacterial bioluminescence into infected cells and allows detection of low numbers of viable target bacteria. The combination of speed, sensitivity, and specificity is superior to previously used diagnostic methods. In conclusion, genetic engineering could improve the properties of phage Y2 toward better killing efficacy and sensitive detection of E. amylovora cells.de_CH
dc.language.isoende_CH
dc.publisherAmerican Society for Microbiologyde_CH
dc.relation.ispartofApplied and Environmental Microbiologyde_CH
dc.rightsLicence according to publishing contractde_CH
dc.subjectBacteriophagede_CH
dc.subjectDepolymerasede_CH
dc.subjectFire blightde_CH
dc.subjectLuciferasede_CH
dc.subjectRecombinant phagede_CH
dc.subjectReporterde_CH
dc.subjectReporter phagede_CH
dc.subjectBacteriophagesde_CH
dc.subjectErwinia amylovorade_CH
dc.subjectViral gene expression regulationde_CH
dc.subjectGenetic engineeringde_CH
dc.subjectMalusde_CH
dc.subjectPlant diseasesde_CH
dc.subjectViral proteinsde_CH
dc.subjectVirulencede_CH
dc.subject.ddc570: Biologiede_CH
dc.titleEngineering of bacteriophages Y2::dpoL1-C and Y2::luxAB for efficient control and rapid detection of the fire blight pathogen, Erwinia amylovorade_CH
dc.typeBeitrag in wissenschaftlicher Zeitschriftde_CH
dcterms.typeTextde_CH
zhaw.departementLife Sciences und Facility Managementde_CH
zhaw.organisationalunitInstitut für Lebensmittel- und Getränkeinnovation (ILGI)de_CH
dc.identifier.doi10.1128/AEM.00341-17de_CH
dc.identifier.pmid28389547de_CH
zhaw.funding.euNode_CH
zhaw.issue12de_CH
zhaw.originated.zhawYesde_CH
zhaw.pages.starte00341-17de_CH
zhaw.publication.statuspublishedVersionde_CH
zhaw.volume83de_CH
zhaw.publication.reviewPeer review (Publikation)de_CH
zhaw.webfeedMikrobiologiede_CH
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Born, Y., Fieseler, L., Thöny, V., Leimer, N., Duffy, B., & Loessner, M. J. (2017). Engineering of bacteriophages Y2::dpoL1-C and Y2::luxAB for efficient control and rapid detection of the fire blight pathogen, Erwinia amylovora. Applied and Environmental Microbiology, 83(12), e00341–17. https://doi.org/10.1128/AEM.00341-17
Born, Y. et al. (2017) ‘Engineering of bacteriophages Y2::dpoL1-C and Y2::luxAB for efficient control and rapid detection of the fire blight pathogen, Erwinia amylovora’, Applied and Environmental Microbiology, 83(12), pp. e00341–17. Available at: https://doi.org/10.1128/AEM.00341-17.
Y. Born, L. Fieseler, V. Thöny, N. Leimer, B. Duffy, and M. J. Loessner, “Engineering of bacteriophages Y2::dpoL1-C and Y2::luxAB for efficient control and rapid detection of the fire blight pathogen, Erwinia amylovora,” Applied and Environmental Microbiology, vol. 83, no. 12, pp. e00341–17, Jun. 2017, doi: 10.1128/AEM.00341-17.
BORN, Yannick, Lars FIESELER, Valentin THÖNY, Nadja LEIMER, Brion DUFFY und Martin J. LOESSNER, 2017. Engineering of bacteriophages Y2::dpoL1-C and Y2::luxAB for efficient control and rapid detection of the fire blight pathogen, Erwinia amylovora. Applied and Environmental Microbiology. Juni 2017. Bd. 83, Nr. 12, S. e00341–17. DOI 10.1128/AEM.00341-17
Born, Yannick, Lars Fieseler, Valentin Thöny, Nadja Leimer, Brion Duffy, and Martin J. Loessner. 2017. “Engineering of Bacteriophages Y2::dpoL1-C and Y2::luxAB for Efficient Control and Rapid Detection of the Fire Blight Pathogen, Erwinia Amylovora.” Applied and Environmental Microbiology 83 (12): e00341–17. https://doi.org/10.1128/AEM.00341-17.
Born, Yannick, et al. “Engineering of Bacteriophages Y2::dpoL1-C and Y2::luxAB for Efficient Control and Rapid Detection of the Fire Blight Pathogen, Erwinia Amylovora.” Applied and Environmental Microbiology, vol. 83, no. 12, June 2017, pp. e00341–17, https://doi.org/10.1128/AEM.00341-17.


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