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Copper resistance genes from different xanthomonads and citrus epiphytic bacteria confer resistance to Xanthomonas citri subsp. citri

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Abstract

Genetic exchange is considered to be an important process in the selective adaptation of microorganisms to shifting and challenging environmental conditions. As a consequence of the copious use of copper bactericides, many species of plant pathogenic bacteria, including Xanthomonas citri subsp. citri (Xcc), have developed resistance to copper. This study assesses whether copper resistant (CuR) strains of other Xanthomonas species and citrus epiphytic bacteria pose a risk for the development of copper resistance in Xcc. CuR epiphytic bacteria were isolated on MGY agar from citrus leaves collected in two citrus groves treated with copper bactericides in Florida. Horizontal gene transfer of copper resistance genes was investigated within different Xanthomonas species and from citrus epiphytic bacteria to Xanthomonas. CuR epiphytic bacteria from citrus were screened for the presence of copper resistance genes homologous to copL, copA and copB genes from Xcc and characterized regarding tolerance to copper. Copper resistance determinants from a citrus epiphytic strain of Stenotrophomonas maltophilia (Stm) were cloned and expressed in Xcc and other Xanthomonas strains. Copper resistance genes in Xcc were determined to be present on a large (~300 kb) conjugative plasmid. Cu resistance was transferred via conjugation from two copper resistant citrus strains, Xcc and X. alfalfae subsp. citrumelonis (Xac), and two tomato pathogens, X. euvesicatoria (Xe) and X. perforans (Xp), to Xcc. PCR analysis revealed that two CuR strains from citrus, an epiphytic Xanthomonas ssp. and a strain of Stm, harboured homologs of the copper resistance genes found in CuR Xcc. The introduction of copLAB gene cluster from Stm into different xanthomonads conferred copper resistance to sensitive strains of Xcc, Xac, Xe and Xp. Based on these results there is a low, but significant, likelihood of horizontal gene transfer of copper resistance genes from other xanthomonads or epiphytic bacteria to Xcc in nature.

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Acknowledgements

We are grateful for support of Franklin Behlau by the Hunt Brothers Fellowship and to the Citrus Research and Development Foundation for grant support.

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  1. Franklin Behlau

    Present address: Fundecitrus–Fundo de Defesa da Citricultura, Av. Adhemar Pereira de Barros, 201, 14807-040, Araraquara, SP, Brazil

Authors and Affiliations

  1. Department of Plant Pathology, University of Florida, Gainesville, FL, 32611, USA

    Franklin Behlau & Jeffrey B. Jones

  2. Instituto Nacional de Tecnología Agropecuaria, Estación Experimental Agropecuaria Bella Vista, Casilla de Correo No. 5, 3432, Bella Vista, Corrientes, Argentina

    Blanca I. Canteros

  3. Citrus Research and Education Center, Soil and Water Science Department, University of Florida, 700 Experiment Road, Lake Alfred, FL, 33850, USA

    James H. Graham

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  1. Franklin Behlau
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Correspondence to James H. Graham.

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Behlau, F., Canteros, B.I., Jones, J.B. et al. Copper resistance genes from different xanthomonads and citrus epiphytic bacteria confer resistance to Xanthomonas citri subsp. citri . Eur J Plant Pathol 133, 949–963 (2012). https://doi.org/10.1007/s10658-012-9966-8

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