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Electrochemical protoplast fusion in citrus

  • Genetic Transformation and Hybridization
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Abstract

We report here the development of a novel protoplast fusion method for citrus somatic hybridization. This new procedure, which we have named electrochemical protoplast fusion, is based on chemically induced protoplast aggregation, using a low concentration of polyethylene glycol, and DC pulse-promoted membrane fusion. Based on the results of nucleus and mitochondria molecular analyses, we were successful in using this method to regenerate both symmetric somatic hybrids and cybrids. Various parameters, including pulse intensity, pulse length, and composition of the fusion media, were tested, and the optimum fusion condition selected consisted of two 100-μs pulses of 1,500 V cm−1. Our conclusion is that electrochemical fusion is a reliable and reproducible method that combines the best features of both the chemical and electrical methods, thereby promoting cell division and high embryogenesis rates of the fused cells. It represents a new approach to citrus somatic hybridization. Various interesting features of this new approach are presented and discussed.

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Acknowledgments

We would like to thank Dr. Carmen Brisa (Universitat de Valencia) for help and directions, Dr. Christine Chase (University of Florida), who kindly provide us with the mitochondrial probe, Dr. Emilio Carbonell (IVIA) for the statistical analysis, and Dr. Cova Arias (Auburn University) for reading of the manuscript. This work was supported by INIA (National Institute for Agricultural Research) Grant no. RTA-02-01

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Authors and Affiliations

  1. Departamento de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias, Apartado Oficial 46113, Moncada-Valencia, Spain

    O. Olivares-Fuster, N. Duran-Vila & L. Navarro

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  1. O. Olivares-Fuster
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  2. N. Duran-Vila
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  3. L. Navarro
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Correspondence to L. Navarro.

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Communicated by W. Harwood

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Olivares-Fuster, O., Duran-Vila, N. & Navarro, L. Electrochemical protoplast fusion in citrus. Plant Cell Rep 24, 112–119 (2005). https://doi.org/10.1007/s00299-005-0916-1

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  • DOI: https://doi.org/10.1007/s00299-005-0916-1

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