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Open AccessArticle

Molecular Profile of Phage Infection: A Novel Approach for the Characterization of Erwinia Phages through qPCR

1
Centre for Biotechnology, Brock University, St. Catharines, ON L2S 3A1, Canada
2
Agriculture and Agri-Food Canada, Vineland Station, ON L0R 2E0, Canada
3
Department of Biological Sciences, Brock University, St. Catharines, ON L2S 3A1, Canada
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(2), 553; https://doi.org/10.3390/ijms21020553
Received: 3 December 2019 / Revised: 10 January 2020 / Accepted: 12 January 2020 / Published: 15 January 2020
(This article belongs to the Special Issue Bacteriophage—Molecular Studies)
Due to the emergence of antibiotic resistance, phage-mediated biocontrol has become an attractive alternative for pathogen management in agriculture. While the infection characteristics of many phages can be adequately described using plaque assays and optical density, the results from phages of the apple pathogen Erwinia amylovora have low reproducibility with these techniques. Using quantitative real-time PCR (qPCR), the stage of the lytic cycle was determined through a combination of chloroform-based sampling, centrifugation, and DNase treatment. Monitoring the transition of phage genomes through the lytic cycle generates a molecular profile from which phage infection characteristics such as adsorption rate and burst size can be determined. To our knowledge, this is the first report of qPCR being used to determine these infection parameters. The characteristics of four different genera of Erwinia phages were determined. The phage ΦEa46-1-A1 was able to adsorb at a rate up to 6.6 times faster than ΦEa35-70 and ΦEa9-2. The low enrichment titer of ΦEa9-2 was shown to be due to the absence of lysis. The ΦEa46-1-A1 and ΦEa21-4 phages had the highest productivity, with burst sizes of 57 virions in 38 min and 185 virions in 98 min, respectively, suggesting these genera would make stronger candidates for the phage-mediated biocontrol of E. amylovora. View Full-Text
Keywords: bacteriophage; Erwinia amylovora; Pantoea agglomerans; biocontrol; adsorption; burst size; latent period bacteriophage; Erwinia amylovora; Pantoea agglomerans; biocontrol; adsorption; burst size; latent period
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Parcey, M.; Gayder, S.; Castle, A.J.; Svircev, A.M. Molecular Profile of Phage Infection: A Novel Approach for the Characterization of Erwinia Phages through qPCR. Int. J. Mol. Sci. 2020, 21, 553.

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