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The potential of bacteriophages to control Xanthomonas campestris pv. campestris at different stages of disease development
Authors:Dominique Holtappels  Kiandro J Fortuna  Lauren Moons  Nand Broeckaert  Léon E Bäcker  Sofie Venneman  Sofie Rombouts  Louis Lippens  Steve Baeyen  Sabien Pollet  Jean-Paul Noben  Frank Oechslin  Marta Vallino  Abram Aertsen  Martine Maes  Johan Van Vaerenbergh  Rob Lavigne  Jeroen Wagemans
Institution:1. Laboratory of Gene Technology, KU Leuven, Leuven, Belgium;2. Laboratory of Food Microbiology, KU Leuven, Leuven, Belgium;3. Proefstation voor de groenteteelt, Sint-Katelijne-Waver, Belgium;4. Laboratory of Gene Technology, KU Leuven, Leuven, Belgium

Flanders Research Institute for Agriculture, Fisheries and Food, Ghent, Belgium;5. Provinciaal Proefcentrum voor de Groenteteelt Oost-Vlaanderen, Kruishoutem, Belgium;6. Flanders Research Institute for Agriculture, Fisheries and Food, Ghent, Belgium;7. Inagro, Beitem, Belgium;8. Biomedical Research Institute and Transnational University Limburg, Hasselt University, Hasselt, Belgium;9. Department of Fundamental Microbiology, University of Lausanne, Lausanne, Switzerland;10. Institute for Sustainable Plant Protection, National Research Counsil of Italy, Turin, Italy;11. Independent Researcher, Ghent, Belgium

Abstract:Xanthomonas campestris pv. campestris (Xcc) is a vascular pathogen that invades the xylem of Brassica crops. Current chemical and antibiotics-based control measures for this bacterium are unsustainable and inefficient. After establishing a representative collection of Xcc strains, we isolated and characterized bacteriophages from two clades of phages to assess their potential in phage-based biocontrol. The most promising phages, FoX2 and FoX6, specifically recognize (lipo) polysaccharides, associated with the wxc gene cluster, on the surface of the bacterial cell wall. Next, we determined and optimized the applicability of FoX2 and FoX6 in an array of complementary bioassays, ranging from seed decontamination to irrigation- and spray-based applications. Here, an irrigation-based application showed promising results. In a final proof-of-concept, a CaCl2-formulated phage cocktail was shown to control the outbreak of Xcc in the open field. This comprehensive approach illustrates the potential of phage biocontrol of black rot disease in Brassica and serves as a reference for the broader implementation of phage biocontrol in integrated pest management strategies.
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