Characterization of Novel Bacteriophages for Biocontrol of Plant Pathogenic Bacteria
DOI:
https://doi.org/10.5281/zenodo.22687887Keywords:
bacteriophage, Ralstonia solanacearum, biocontrol, bacterial wilt, phage cocktail, tomato, lytic phageAbstract
Bacterial wilt, caused by the soilborne pathogen Ralstonia solanacearum, remains one of the most destructive and
management-resistant bacterial diseases of tomato and other solanaceous crops in warm, humid production
regions, and no chemical control offers reliable field efficacy once the pathogen is established in soil. We report
the isolation, characterisation and greenhouse biocontrol evaluation of three novel lytic bacteriophages,
designated RsP7, RsP12 and RsP19, recovered from wilt-affected tomato field soil and irrigation water in a
case-study system centred on a single commercial glasshouse operation with a recurrent bacterial wilt problem.
Transmission electron microscopy placed all three phages in the class Caudoviricetes, with RsP7 and RsP19
showing Myoviridae-like contractile tails and RsP12 showing a Podoviridae-like short tail. Whole-genome
sequencing yielded double-stranded DNA genomes of 66.2-72.8 kb with no genes matching curated toxin,
integrase or antibiotic-resistance reference databases, supporting their suitability for field application under a
strictly lytic-phage biocontrol strategy. Host-range testing against 24 R. solanacearum isolates spanning
phylotypes I and IIB collected from the same production region showed partially complementary lysis patterns, and
a three-phage cocktail combining RsP7, RsP12 and RsP19 lysed 22 of 24 isolates (91.7%), compared with a
maximum of 15 of 24 (62.5%) for any single phage. In a greenhouse pot trial using naturally infested field soil,
cocktail-treated tomato plants showed 58% wilt incidence at 21 days post-transplant versus 89% in untreated
controls, and time-to-wilt onset was delayed by a median of 6 days. These results support this three-phage
cocktail as a promising candidate for field-scale biocontrol evaluation against bacterial wilt in this production
system, while underlining that phage cocktail performance should be characterised against a genotypically
representative local pathogen population rather than a single reference strain before any biocontrol product is
deployed more broadly.
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