title: The use of stable and unstable green fluorescent proteins for studies in two bacterial models: Agrobacterium tumefaciens and Xanthomonas campestris pv. campestris.
abstract: Fluorescent proteins have been used to track plant pathogens to understand their host interactions. To be useful, the transgenic pathogens must present similar behaviour than the wild-type isolates. Herein, a GFP marker was used to transform two plant pathogenic bacteria, Agrobacterium and Xanthomonas, to localize and track the bacteria during infection. The transgenic bacteria were evaluated to determine whether they showed the same fitness than the wild-type strains or whether the expression of the GFP protein interfered in the bacterial activity. In Agrobacterium, the plasmid used for transformation was stable in the bacteria and the strain kept the virulence, while Xanthomonas was not able to conserve the plasmid and transformed strains showed virulence variations compared to wild-type strains. Although marking bacteria with GFP to track infection in plants is a common issue, works to validate the transgenic strains and corroborate their fitness are not usual. Results, presented here, confirm the importance of proper fitness tests on the marked strains before performing localization assays, to avoid underestimation of the microbe population or possible artificial effects in its interaction with the plant.
authors.affiliation: Laboratorio de Bacteriología. Departamento de Protección Vegetal, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain.
authors.forename: Pilar
authors.initials: P
authors.lastname: Sabuquillo
authors.affiliation: Laboratorio de Bacteriología. Departamento de Protección Vegetal, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain.
authors.forename: Adela
authors.initials: A
authors.lastname: Gea
authors.affiliation: Área de Genética, Facultad de Ciencias, Instituto de Hortofruticultura Subtropical y Mediterránea 'La Mayora', Universidad de Málaga-Consejo Superior de Investigaciones Científicas (IHSM-UMA-CSIC), Málaga, Spain.|Instituto de Agrobiotecnología, CSIC-UPNA, Gobierno de Navarra, 31192, Mutilva, Navarra, Spain.
authors.forename: Isabel M
authors.initials: IM
authors.lastname: Matas
authors.affiliation: Área de Genética, Facultad de Ciencias, Instituto de Hortofruticultura Subtropical y Mediterránea 'La Mayora', Universidad de Málaga-Consejo Superior de Investigaciones Científicas (IHSM-UMA-CSIC), Málaga, Spain.
authors.forename: Cayo
authors.initials: C
authors.lastname: Ramos
authors.affiliation: Laboratorio de Bacteriología. Departamento de Protección Vegetal, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain. cubero@inia.es.
authors.forename: Jaime
authors.identifier: http://orcid.org/0000-0002-4314-857X
authors.initials: J
authors.lastname: Cubero
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics* / Q000472:pathogenicity; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000235:genetics; D008954:Models, Biological; D030781:Organisms, Genetically Modified; D010935:Plant Diseases / Q000382:microbiology; D010944:Plants / Q000382:microbiology; D010957:Plasmids / Q000235:genetics; D014169:Transformation, Bacterial; D014774:Virulence; D016959:Xanthomonas campestris / Q000235:genetics* / Q000472:pathogenicity
chemical_list: D049452:Green Fluorescent Proteins
keywords: Bacterial fitness; Black rot; Crown gall; GFP; Plant disease; Plant pathology