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Endophytic Methylobacterium extorquens expresses a heterologous β-1,4-endoglucanase A (EglA) in Catharanthus roseus seedlings, a model host plant for Xylella fastidiosa

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Abstract

Based on the premise of symbiotic control, we genetically modified the citrus endophytic bacterium Methylobacterium extorquens, strain AR1.6/2, and evaluated its capacity to colonize a model plant and its interaction with Xylella fastidiosa, the causative agent of Citrus Variegated Chlorosis (CVC). AR1.6/2 was genetically transformed to express heterologous GFP (Green Fluorescent Protein) and an endoglucanase A (EglA), generating the strains ARGFP and AREglA, respectively. By fluorescence microscopy, it was shown that ARGFP was able to colonize xylem vessels of the Catharanthus roseus seedlings. Using scanning electron microscopy, it was observed that AREglA and X. fastidiosa may co-inhabit the C. roseus vessels. M. extorquens was observed in the xylem with the phytopathogen X. fastidiosa, and appeared to cause a decrease in biofilm formation. AREglA stimulated the production of resistance protein, catalase, in the inoculated plants. This paper reports the successful transformation of AR1.6/2 to generate two different strains with a different gene each, and also indicates that AREglA and X. fastidiosa could interact inside the host plant, suggesting a possible strategy for the symbiotic control of CVC disease. Our results provide an enhanced understanding of the M. extorquens—X. fastidiosa interaction, suggesting the application of AR1.6/2 as an agent of symbiotic control.

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Acknowledgments

This work was supported by a grant (Proc. no. 98/16262-2) fellowship to M.C.Q. (Proc. no. 2005/53748-6), P.B.R. (Proc. no. 00/13800-5) and W.L.A (Proc. no. 00/10699-1) from FAPESP. We also thank Dr. Elliot W. Kitajima (NAP/MEA/ESALQ) for providing microscopy facilities, Dr. Marianne Figueira (Biotechnology Research Institute, Canada), Dr. Christopher Marxs and Dr. Mary Lidstrom (University of Washington, USA) for discussions and plasmids.

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Authors and Affiliations

  1. Department of Phytopathology, University of Passo Fundo (UPF), BR 285, PO BOX 611, Passo Fundo, RS, 99052-900, Brazil

    Antônio Sérgio Ferreira Filho

  2. Department of Genetics, Escola Superior de Agricultura “Luiz de Queiroz”, University of São Paulo, 11 Pádua Dias Av., PO BOX 83, Piracicaba, SP, 13400-970, Brazil

    Maria Carolina Quecine, Andréa Cristina Bogas & João Lúcio Azevedo

  3. Laboratory of Biotechnology, Centro de Energia Nuclear na Agricultura, University of São Paulo, 303 Centenário Av., PO BOX 96, Piracicaba, SP, 13416 000, Brazil

    Priscilla de Barros Rossetto

  4. Center of Science Tecnology of Earh and Sea, University of Vale do Itajaí, 458 Uruguai Street, 14, Itajaí, SC, 88302-202, Brazil

    Andre Oliveira de Souza Lima

  5. Institute of Natural Science, Federal University of Alfenas, Alfenas, MG, 37130-000, Brazil

    Paulo Teixeira Lacava

  6. Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, Av. Prof. Lineu Prestes, 1374 - Ed. Biomédicas II, Cidade Universitária, São Paulo, SP, 05508-900, Brazil

    Welington Luiz Araújo

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  1. Antônio Sérgio Ferreira Filho
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  2. Maria Carolina Quecine
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  5. Andre Oliveira de Souza Lima
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Correspondence to Welington Luiz Araújo.

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Ferreira Filho, A.S., Quecine, M.C., Bogas, A.C. et al. Endophytic Methylobacterium extorquens expresses a heterologous β-1,4-endoglucanase A (EglA) in Catharanthus roseus seedlings, a model host plant for Xylella fastidiosa . World J Microbiol Biotechnol 28, 1475–1481 (2012). https://doi.org/10.1007/s11274-011-0949-2

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  • DOI: https://doi.org/10.1007/s11274-011-0949-2

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