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Enhanced resistance to citrus canker in transgenic sweet orange expressing the sarcotoxin IA gene

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Abstract

Citrus canker, caused by the bacterial pathogen Xanthomonas citri subp. Citri (Xcc), is a serious disease reported in most citrus-producing areas around the world. Although different levels of field resistance to citrus canker have been reported in sweet oranges, they are usually not sufficient to provide adequate control of the disease. Ectopic over-expression of antibacterial genes is one of the potential strategies to increase plant resistance to bacterial diseases. Previous in vitro results showed that sarcotoxin IA, an antimicrobial peptide isolated from the flesh fly (Sarcophaga peregrina), can be efficient to control different plant pathogenic bacteria, including Xcc. Transgenic “Pera” sweet orange (Citrus sinensis [L.] Osbeck) plants constitutively expressing the sarcotoxin IA peptide fused to the PR1a signal peptide from Nicotiana tabacum for secretion in the intercellular space were obtained by Agrobacterium-mediated transformation using thin sections of mature explants. Citrus canker resistance evaluation in leaves of transgenic and non-transgenic plants was performed through inoculations with Xcc by infiltration and spraying. The Xcc population was up to 2 log unit lower in leaves of the transgenic plants compared to those of non-transgenic controls. Incidence of canker lesions was significantly higher in non-transformed controls (>10 lesions/cm2) than in the transgenic plants (<5 lesions/cm2) after injection infiltration or spraying with Xcc inoculum. Accumulation of sarcotoxin IA peptide in sweet orange tissue did not cause any deleterious effects on the growth and development of the transgenic plants, indicating this approach is suitable to provide resistance to citrus canker.

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Acknowledgements

The authors gratefully thanks to Dr. Yuko Ohashi (Plant-Microbe Interactions Research Unit, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan) for providing the pST10 plasmid. We also thank Suely A. Kudo and Luciana Meneguin for technical assistance. We thank Dr. Nelson A. Wulff, Dr. Leandro Peña and Dr. James Graham for critical reading of this manuscript. This work received financial support from CNPq, Fundação Araucária and Fundo de Defesa da Citricultura – FUNDECITRUS. L. G. E. Vieira and L. F. P. Pereira are CNPq research fellows.

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  1. Adilson K. Kobayashi & Hugo Bruno C. Molinari

    Present address: Genetics and Biotechnology Laboratory, Embrapa Agroenergy, Brasília, DF, 70770-901, Brazil

  2. Luiz Gonzaga E. Vieira

    Present address: Biotechnology Laboratory, UNOESTE, Rodovia Raposo Tavares, Km 572 - Limoeiro, Presidente Prudente, SP, 19.067-175, Brazil

  3. João Carlos Bespalhok Filho

    Present address: Universidade Federal do Paraná, R. dos Funcionários, 1540, Curitiba, PR, 80035-050, Brazil

  4. Viviani V. Marques

    Present address: Biothecnology Laboratory, Fundo de Defesa da Citricultura – FUNDECITRUS, Av. Dr. Adhemar Pereira de Barros, 201 –, Araraquara, SP, 14.807-040, Brazil

Authors and Affiliations

  1. Instituto Agronômico do Paraná - IAPAR, Rodovia Celso Garcia Cid, km 375, Londrina, PR, 86001-970, Brazil

    Adilson K. Kobayashi, Luiz Gonzaga E. Vieira, João Carlos Bespalhok Filho, Rui Pereira Leite Jr & Hugo Bruno C. Molinari

  2. Embrapa Café, Biothecnology Laboratory – IAPAR, Rod. Celso Garcia Cid Km 375, Londrina, PR, 86001-970, Brazil

    Luiz Filipe P. Pereira

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

    Viviani V. Marques

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Kobayashi, A.K., Vieira, L.G.E., Bespalhok Filho, J.C. et al. Enhanced resistance to citrus canker in transgenic sweet orange expressing the sarcotoxin IA gene. Eur J Plant Pathol 149, 865–873 (2017). https://doi.org/10.1007/s10658-017-1234-5

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