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Isolation, characterization, and evaluation of putative new bacteriophages for controlling bacterial spot on tomato in Brazil

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Abstract

Bacterial spot is a highly damaging tomato disease caused by members of several species of the genus Xanthomonas. Bacteriophages have been studied for their potential use in the biological control of bacterial diseases. In the current study, bacteriophages were obtained from soil and tomato leaves in commercial fields in Brazil with the aim of obtaining biological control agents against bacterial spot. Phage isolation was carried out by co-cultivation with isolates of Xanthomonas euvesicatoria pv. perforans, which was prevalent in the collection areas. In a host range evaluation, none of the phage isolates was able to induce a lytic cycle in all of the bacterial isolates tested. In in vivo tests, treatment of susceptible bacterial isolates with the corresponding phage prior to application to tomato plants led to a reduction in the severity of the resulting disease. The level of disease control provided by phage application was equal to or greater than that achieved using copper hydroxide. Electron microscopy analysis showed that all of the phages had similar morphology, with head and tail structures similar to those of viruses belonging to the class Caudoviricetes. The presence of short, non-contractile tubular tails strongly suggested that these phages belong to the family Autographiviridae. This was confirmed by phylogenetic analysis, which further revealed that they all belong to the genus Pradovirus. The phages described here are closely related to each other and potentially belong to a new species within the genus. These phages will be evaluated in future studies against other tomato xanthomonad strains to assess their potential as biological control agents.

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Acknowledgments

The authors would like to thank the Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF, PRONEX 0193.001197/2016 and Demanda Espontânea 09/2022), the Fundação de Amparo à Pesquisa do Estado de Goiás (FAPEG, DOCFIX 08/2018), and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, UNIVERSAL 28/2018) for financial support of this study. AKI-N is a CNPq fellow.

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Authors and Affiliations

  1. Programa de Pós-Graduação em Olericultura, Instituto Federal Goiano (IF Goiano), Morrinhos, GO, Brazil

    Dayane Maria de Sousa, Raphael Barboza Rosa, Jaqueline Kiyomi Yamada & Nadson de Carvalho Pontes

  2. Laboratory of Baculovirus, Cell Biology Department, University of Brasília (UnB), Campus Universitário Darcy Ribeiro, Brasília, DF, Brazil

    Luis Janssen, Aline Belmok, Roberto Franco Teixeira Corrêa, Miguel de Souza Andrade & Bergmann Morais Ribeiro

  3. Laboratory of Virology, Embrapa Vegetables, Brasilia, DF, Brazil

    Alice Kazuko Inoue-Nagata

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  1. Dayane Maria de Sousa
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705_2023_5846_MOESM1_ESM.docx

Supplementary Figure 1: Representative electron microscope micrograph of Phage PL4 stained with uranyl acetate. The image inset shows the head-tail morphology typical of those members of order Caudovirales, albeit the virion presents a short tail.

705_2023_5846_MOESM2_ESM.docx

Supplementary Figure 2: Proteome-based phylogenetic relationships among Xanthomonas-infecting bacteriophages. Lineages marked with stars correspond to the phages described in the current study.

705_2023_5846_MOESM3_ESM.docx

Supplementary Figure 3: Syntheny of whole genomes of Xanthomonas phages described in the current study. The genomes are highly co-linear, other than the lysis genes region, which occurs at the 5? edge of the GF1 genome. Moreover, the F5 genome has three consecutive copies (in gray) of tail-coding genes, which may confer a higher host range to its virus.

Supplementary Material 4

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de Sousa, D.M., Janssen, L., Rosa, R.B. et al. Isolation, characterization, and evaluation of putative new bacteriophages for controlling bacterial spot on tomato in Brazil. Arch Virol 168, 222 (2023). https://doi.org/10.1007/s00705-023-05846-y

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