{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T10:12:55Z","timestamp":1787479975581,"version":"build-2736575974"},"reference-count":52,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2022,6,24]],"date-time":"2022-06-24T00:00:00Z","timestamp":1656028800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Comisi\u00f3n Nacional de Investigaci\u00f3n Cient\u00edfica y Tecnol\u00f3gica (CONICYT, Chile)","award":["3180660"],"award-info":[{"award-number":["3180660"]}]},{"name":"Comisi\u00f3n Nacional de Investigaci\u00f3n Cient\u00edfica y Tecnol\u00f3gica (CONICYT, Chile)","award":["RGPIN-2018-05942"],"award-info":[{"award-number":["RGPIN-2018-05942"]}]},{"name":"NSERC-Discovery grant","award":["3180660"],"award-info":[{"award-number":["3180660"]}]},{"name":"NSERC-Discovery grant","award":["RGPIN-2018-05942"],"award-info":[{"award-number":["RGPIN-2018-05942"]}]},{"name":"Genome Atlantic and Atlantic Canada Opportunities Agency (ACOA)","award":["3180660"],"award-info":[{"award-number":["3180660"]}]},{"name":"Genome Atlantic and Atlantic Canada Opportunities Agency (ACOA)","award":["RGPIN-2018-05942"],"award-info":[{"award-number":["RGPIN-2018-05942"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Viruses"],"abstract":"<jats:p>Xanthomonas arboricola pv. juglandis (hereafter X. juglandis) is the etiological agent of walnut blight, the most important bacterial disease affecting walnut production worldwide. Currently, the disease is treated mainly with copper-derived compounds (e.g., CuSO4) despite the evidence of genetic resistance in these strains. Regarding the effectiveness and sustainability, the use of a bacteriophage appears to be a biocontrol alternative to reduce X. juglandis load and symptomatology of walnut blight. Here, the phages f20-Xaj, f29-Xaj, and f30-Xaj were characterized, and their effectiveness in walnut orchards against walnut blight was determined. These bacteriophages showed a specific lytic infection in X. juglandis strains isolated from Chile and France. Phylogenetic analysis of the complete genome of f20-Xaj and f30-Xaj indicates that these phages belong to the Pradovirus genus. In the field, the cocktail of these bacteriophages showed similar effectivity to CuSO4 in the reduction of incidence and severity in walnut tissue. Moreover, the bacterial load of X. juglandis was significantly reduced in the presence of bacteriophages in contrast to a CuSO4 treatment. These results show that the use of bacteriophages can be an alternative to combat the symptoms of walnut blight caused by X. juglandis.<\/jats:p>","DOI":"10.3390\/v14071380","type":"journal-article","created":{"date-parts":[[2022,6,26]],"date-time":"2022-06-26T22:50:23Z","timestamp":1656283823000},"page":"1380","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["Characterization of Xanthomonas arboricola pv. juglandis Bacteriophages against Bacterial Walnut Blight and Field Evaluation"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1290-7286","authenticated-orcid":false,"given":"Julio","family":"Retamales","sequence":"first","affiliation":[{"name":"Instituto de Ciencias Naturales, Facultad de Medicina Veterinaria y Agronom\u00eda, Universidad de las Am\u00e9ricas, Vi\u00f1a del Mar 2520000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pablo","family":"N\u00fa\u00f1ez","sequence":"additional","affiliation":[{"name":"Agroadvance SpA, Pe\u00f1aflor 9750000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Romina","family":"Alvarado","sequence":"additional","affiliation":[{"name":"Agroadvance SpA, Pe\u00f1aflor 9750000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4485-0918","authenticated-orcid":false,"given":"Erick D. M.","family":"Campan","sequence":"additional","affiliation":[{"name":"Laboratoire \u00c9cologie Fonctionnelle et Environnement, Universit\u00e9 de Toulouse, CNRS, 31062 Toulouse, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2854-6454","authenticated-orcid":false,"given":"Thierry","family":"Otto","sequence":"additional","affiliation":[{"name":"Laboratoire \u00c9cologie Fonctionnelle et Environnement, Universit\u00e9 de Toulouse, CNRS, 31062 Toulouse, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristopher","family":"Segovia","sequence":"additional","affiliation":[{"name":"Marine Microbial Pathogenesis and Vaccinology Laboratory, Department of Ocean Sciences, Memorial University of Newfoundland, St. John\u2019s, NL A1C 5S7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-1390-2809","authenticated-orcid":false,"given":"Ignacio","family":"Vasquez","sequence":"additional","affiliation":[{"name":"Marine Microbial Pathogenesis and Vaccinology Laboratory, Department of Ocean Sciences, Memorial University of Newfoundland, St. John\u2019s, NL A1C 5S7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7701-3147","authenticated-orcid":false,"given":"Javier","family":"Santander","sequence":"additional","affiliation":[{"name":"Marine Microbial Pathogenesis and Vaccinology Laboratory, Department of Ocean Sciences, Memorial University of Newfoundland, St. John\u2019s, NL A1C 5S7, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,24]]},"reference":[{"key":"ref_1","first-page":"23","article-title":"Characterization of Xanthomonas arboricola pv. juglandis isolated from walnuts in Lithuania","volume":"94","author":"Burokiene","year":"2012","journal-title":"J. Plant Pathol."},{"key":"ref_2","first-page":"79","article-title":"Bacterial diseases of walnut and hazelnut and genetic resources","volume":"92","author":"Frutos","year":"2010","journal-title":"J. Plant Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1163","DOI":"10.1094\/PHYTO-01-14-0001-R","article-title":"Colonization of dormant walnut buds by Xanthomonas arboricola pv. juglandis is predictive of subsequent disease","volume":"104","author":"Lindow","year":"2014","journal-title":"Phytopathology"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"253","DOI":"10.15835\/nsb729620","article-title":"Assessment of bacterial blight tolerance of persian walnut based on immature nut test","volume":"7","author":"Bandi","year":"2015","journal-title":"Not. Sci. Biol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"326452","DOI":"10.1155\/2012\/326452","article-title":"Advances in bacteriophage mediated control of plant pathogens","volume":"2012","author":"Frampton","year":"2012","journal-title":"Int. J. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"259","DOI":"10.17660\/ActaHortic.1993.311.33","article-title":"Copper resistance of Xanthomonas campestris pv. juglandis in French walnut orchards and its association with conjugative plasmids","volume":"311","author":"Gardan","year":"1993","journal-title":"Acta Hort"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1128\/jb.176.1.173-188.1994","article-title":"Molecular cloning, chromosomal mapping, and sequence-analysis of copper resistance genes from Xanthomonas campestris pv. juglandis: Homology with small blue copper proteins and multicopper oxidase","volume":"176","author":"Lee","year":"1994","journal-title":"J. Bacteriol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s10658-015-0809-2","article-title":"Morphological and genotypic features of Xanthomonas arboricola pv. juglandis populations from walnut groves in Romagna region, Italy","volume":"145","author":"Giovanardi","year":"2016","journal-title":"Eur. J. Plant Pathol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e01160-15","DOI":"10.1128\/genomeA.01160-15","article-title":"Draft genome sequences of four Xanthomonas arboricola pv. juglandis strains associated with walnut blight in Chile","volume":"3","author":"Higuera","year":"2015","journal-title":"Genome Announc."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e01126-15","DOI":"10.1128\/genomeA.01126-15","article-title":"Complete genome sequence of Xanthomonas arboricola pv. juglandis 417, a copper-resistant strain isolated from Juglans regia L.","volume":"3","author":"Pereira","year":"2015","journal-title":"Genome Announc."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e01085-17","DOI":"10.1128\/genomeA.01085-17","article-title":"Draft genome sequence of Xanthomonas arboricola pv. juglandis J303, isolated from infected walnut trees in southern Chile","volume":"5","author":"Retamales","year":"2017","journal-title":"Genome Announc."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e00023-18","DOI":"10.1128\/genomeA.00023-18","article-title":"Complete genome sequence of Xanthomonas arboricola pv. juglandis strain DW3F3, isolated from a Juglans regia L. bacterial blighted fruitlet","volume":"6","author":"Fu","year":"2018","journal-title":"Genome Announc."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Svircev, A., Roach, D., and Castle, A. (2018). Framing the Future with Bacteriophages in Agriculture. Viruses, 10.","DOI":"10.3390\/v10050218"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"34","DOI":"10.3389\/fmicb.2017.00034","article-title":"Bacteriophages and bacterial plant diseases","volume":"8","author":"Buttimer","year":"2017","journal-title":"Front. Microbiol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Stefani, E., Obradovi\u0107, A., Ga\u0161i\u0107, K., Altin, I., Nagy, I.K., and Kov\u00e1cs, T. (2021). Bacteriophage-Mediated Control of Phytopathogenic Xanthomonads: A Promising Green Solution for the Future. Microorganisms, 9.","DOI":"10.3390\/microorganisms9051056"},{"key":"ref_16","first-page":"220","article-title":"Bacteriophages: A potential biocontrol agent against walnut blight (Xanthomonas campestris pv juglandis)","volume":"54","author":"McNeil","year":"2001","journal-title":"N. Z. Plant Prot."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1917","DOI":"10.1007\/s11274-011-0992-z","article-title":"Isolation and characterization of bacteriophages infecting Xanthomonas arboricola pv. juglandis, the causal agent of walnut blight disease","volume":"28","author":"Jordan","year":"2012","journal-title":"World J. Microbiol. Biotechnol."},{"key":"ref_18","first-page":"371","article-title":"Comparative analysis of two bacteriophages of Xanthomonas arboricola pv. juglandis","volume":"6","author":"Frank","year":"2016","journal-title":"Infect. Genet. Evol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e00336-16","DOI":"10.1128\/genomeA.00336-16","article-title":"Complete genome sequences of lytic bacteriophages of Xanthomonas arboricola pv. juglandis","volume":"4","author":"Retamales","year":"2016","journal-title":"Genome Announc."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1016\/j.ygeno.2010.06.001","article-title":"Identification of lytic bacteriophage MmP1, assigned to a new member of T7-like phages infecting Morganella morganii","volume":"96","author":"Zhu","year":"2010","journal-title":"Genomics"},{"key":"ref_21","first-page":"155","article-title":"Detection and identification methods and new tests as developed and used in the framework of cost 873 for bacteria pathogenic to stone fruits and nuts: Xanthomonas arboricola pv. juglandis","volume":"94","author":"Moragrega","year":"2012","journal-title":"J. Plant Pathol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1128\/jb.173.2.697-703.1991","article-title":"16S ribosomal DNA amplification for phylogenetic study","volume":"173","author":"Weisberg","year":"1991","journal-title":"J. Bacteriol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1111\/j.1574-6968.1993.tb06508.x","article-title":"Fast classification of plant-associated bacteria in the Xanthomonas genus","volume":"113","author":"Maes","year":"1993","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.mimet.2011.03.019","article-title":"A duplex-PCR method for species-and pathovar-level identification and detection of the quarantine plant pathogen Xanthomonas arboricola pv. pruni","volume":"86","author":"Pagani","year":"2011","journal-title":"J. Microbiol. Methods"},{"key":"ref_25","first-page":"17","article-title":"Aislamiento y caracterizaci\u00f3n de bacteri\u00f3fagos l\u00edticos contra Salmonella enteritidis y su ensayo sobre Salmonella pullorum","volume":"8","author":"Santander","year":"2002","journal-title":"Acta Microbiol."},{"key":"ref_26","unstructured":"Davis, R., Botstein, D., and Roth, J. (1980). A Manual for Genetic Engineering: Advanced Bacterial Genetics, Cold Spring Harbor Laboratory Press. [2nd ed.]."},{"key":"ref_27","unstructured":"Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press. [2nd ed.]."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1007\/BF00205063","article-title":"Characterization of a virulent Lactobacillus sake phage PWH2","volume":"39","author":"Leuschner","year":"1993","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1085\/jgp.22.3.365","article-title":"The growth of bacteriophage","volume":"22","author":"Ellis","year":"1939","journal-title":"J. Gen. Physiol."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Adams, M.H. (1959). Bacteriophages, Interscience Publishers.","DOI":"10.5962\/bhl.title.6966"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1093\/bib\/bbn017","article-title":"MEGA: A biologist-centric software for evolutionary analysis of DNA and protein sequences","volume":"9","author":"Kumar","year":"2008","journal-title":"Brief. Bioinform."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3389","DOI":"10.1093\/nar\/25.17.3389","article-title":"Gapped BLAST and PSI-BLAST: A new generation of protein database search programs","volume":"25","author":"Altschul","year":"1997","journal-title":"Nucleic Acids Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"6924","DOI":"10.1128\/JB.00831-06","article-title":"Genomic analysis of Pseudomonas aeruginosa phages LKD16 and LKA1: Establishment of the phiKMV subgroup within the T7 supergroup","volume":"188","author":"Ceyssens","year":"2006","journal-title":"J. Bacteriol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1701","DOI":"10.1099\/mic.0.28712-0","article-title":"A generalized transducing phage (phiIF3) for the genomically sequenced Serratia marcescens strain Db11: A tool for functional genomics of an opportunistic human pathogen","volume":"152","author":"Petty","year":"2006","journal-title":"Microbiology"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"7308","DOI":"10.1128\/JB.181.23.7308-7313.1999","article-title":"Molecular Evidence for a new bacteriophage of Borrelia burgdorferi","volume":"181","author":"Eggers","year":"1999","journal-title":"J. Bacteriol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Glover, D., and Hames, B. (1995). Construction of representative genomic DNA libraries using phages lambda replacement vectors. A Practical Approach DNA Cloning, Oxford University Press.","DOI":"10.1093\/oso\/9780199634835.001.0001"},{"key":"ref_37","first-page":"406","article-title":"The neighbor-joining method: A new method for reconstructing phylogenetic trees","volume":"4","author":"Saitou","year":"1987","journal-title":"Mol. Biol. Evol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Nei, M., and Kumar, S. (2000). Molecular Evolution and Phylogenetics, Oxford University Press.","DOI":"10.1093\/oso\/9780195135848.001.0001"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1547","DOI":"10.1093\/molbev\/msy096","article-title":"MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms","volume":"35","author":"Kumar","year":"2018","journal-title":"Mol. Biol. Evol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"537","DOI":"10.1093\/bioinformatics\/bti054","article-title":"Circular genome visualization and exploration using CGView","volume":"21","author":"Stothard","year":"2005","journal-title":"Bioinformatics"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1009","DOI":"10.1093\/bioinformatics\/btr039","article-title":"Easyfig: A genome comparison visualizer","volume":"27","author":"Sullivan","year":"2011","journal-title":"Bioinformatics"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"783","DOI":"10.2307\/2408678","article-title":"Confidence limits on phylogenies: An approach using the bootstrap","volume":"39","author":"Felsenstein","year":"1985","journal-title":"Evolution"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.copbio.2020.08.016","article-title":"The future of phage biocontrol in integrated plant protection for sustainable crop production","volume":"68","author":"Holtappels","year":"2021","journal-title":"Curr. Opin. Biotechnol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1481","DOI":"10.1111\/mpp.13073","article-title":"Xanthomonas arboricola pv. juglandis and pv. corylina: Brothers or distant relatives? Genetic clues, epidemiology, and insights for disease management","volume":"22","author":"Pothier","year":"2021","journal-title":"Mol. Plant Pathol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"65","DOI":"10.31421\/IJHS\/20\/1-2\/1119","article-title":"Comparison of Xanthomonas arboricola pv. juglandis isolates from walnut trees grown in Romania and Hungary","volume":"20","author":"Bandi","year":"2014","journal-title":"Int. J. Hortic. Sci."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"858","DOI":"10.1094\/PDIS-10-16-1481-RE","article-title":"Multiple DNA markers for identification of Xanthomonas arboricola pv. juglandis isolates and its direct detection in plant samples","volume":"101","author":"Fernandes","year":"2017","journal-title":"Plant Dis."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Fernandes, C., Martins, L., Teixeira, M., Blom, J., Pothier, J.F., Fonseca, N.A., and Tavares, F. (2021). Comparative genomics of Xanthomonas euroxanthea and Xanthomonas arboricola pv. juglandis strains isolated from a single walnut host tree. Microorganisms, 9.","DOI":"10.3390\/microorganisms9030624"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"fnw002","DOI":"10.1093\/femsle\/fnw002","article-title":"Host receptors for bacteriophage adsorption","volume":"363","author":"Bertozzi","year":"2016","journal-title":"FEMS Microbio. Lett."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"De Leeuw, M., Baron, M., David, O.B., and Kushmaro, A. (2020). Molecular Insights into Bacteriophage Evolution toward Its Host. Viruses, 12.","DOI":"10.3390\/v12101132"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Sabuquillo, P., and Cubero, J. (2021). Biofilm formation in Xanthomonas arboricola pv. pruni: Structure and Development. Agronomy, 11.","DOI":"10.3390\/agronomy11030546"},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Rosner, D., and Clark, J. (2021). Formulations for bacteriophage therapy and the potential uses of immobilization. Pharmaceuticals, 14.","DOI":"10.3390\/ph14040359"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1565","DOI":"10.1094\/PDIS-03-11-0259","article-title":"Apical necrosis and premature drop of Persian (English) walnut fruit caused by Xanthomonas arboricola pv. juglandis","volume":"95","author":"Moragrega","year":"2011","journal-title":"Plant Dis."}],"container-title":["Viruses"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/7\/1380\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:39:20Z","timestamp":1760139560000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4915\/14\/7\/1380"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,24]]},"references-count":52,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["v14071380"],"URL":"https:\/\/doi.org\/10.3390\/v14071380","relation":{},"ISSN":["1999-4915"],"issn-type":[{"value":"1999-4915","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,6,24]]}}}