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Reaction of transgenic Citrus sinensis plants to Citrus tristeza virus infection by Toxoptera citricida

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Abstract

Transgenic Citrus sinensis ‘Hamlin’ and ‘Valencia’ plants containing Citrus tristeza virus (CTV)-derived sequences were propagated and inoculated with CTV. For propagation, selected buds from transgenic and non-transgenic control plants were grafted onto C. aurantium and C. limonia rootstock plants. CTV inoculation was performed via viruliferous aphids (Toxoptera citricida), and viral detection post-inoculation was performed through DASI-ELISA or RT-qPCR. After four inoculations, none of the transgenic lines tested showed complete resistance. However, viral multiplication was undetectable in some of the propagated clones. These resistant clones mainly came from transgenic ‘Valencia’ sweet orange plants grafted onto C. limonia rootstock containing the pCTV-CS gene construct. Although the tested viral inoculation method represents natural field infection conditions, the results did not differ significantly from those previously reported when the same transgenic lines were bud-graft inoculated. This finding indicates that the difficulties in producing CTV-resistant transgenic citrus lines may be unrelated to the inoculation method. Transgene expression level was quantified by RT-qPCR analysis and it was not possible to relate transgene mRNA level with resistance to the pathogen.

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Acknowledgments

The authors acknowledge Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) and Fundo de Defesa da Citricultura (Fundecitrus) for research financial support. FRM acknowledges Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for doctoral fellowship. FAAMF, JAMR, RH, and BMJM acknowledge CNPq for research fellowship.

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Authors and Affiliations

  1. Escola Superior de Agricultura “Luiz de Queiroz”, Universidade de São Paulo, 13418-900, Piracicaba, SP, Brazil

    Fabiana R. Muniz, Amancio Souza, Francisco de Assis Alves Mourão Filho & Jorge A. M. Rezende

  2. Instituto Biológico de São Paulo, 04014-002, São Paulo, SP, Brazil

    Ricardo Harakava

  3. Instituto de Biologia, Universidade Estadual de Campinas, 13083-970, Campinas, SP, Brazil

    Dagmar R. Stach-Machado

  4. University of Florida, Gainesville, FL, USA

    Vicente J. Febres & Gloria A. Moore

  5. Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, 13400-970, Piracicaba, SP, Brazil

    Beatriz M. J. Mendes

Authors
  1. Fabiana R. Muniz
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  2. Amancio Souza
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  3. Ricardo Harakava
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  4. Francisco de Assis Alves Mourão Filho
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  5. Dagmar R. Stach-Machado
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  6. Jorge A. M. Rezende
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  7. Vicente J. Febres
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  8. Gloria A. Moore
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  9. Beatriz M. J. Mendes
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Correspondence to Beatriz M. J. Mendes.

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Muniz, F.R., Souza, A., Harakava, R. et al. Reaction of transgenic Citrus sinensis plants to Citrus tristeza virus infection by Toxoptera citricida . Eur J Plant Pathol 139, 151–159 (2014). https://doi.org/10.1007/s10658-013-0373-6

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  • DOI: https://doi.org/10.1007/s10658-013-0373-6

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