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Regeneration and characterization of somatic hybrids combining sweet orange and mandarin/mandarin hybrid cultivars for citrus scion improvement

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Abstract

Protoplast fusion between sweet orange and mandarin/mandarin hybrids scion cultivars was performed following the model “diploid embryogenic callus protoplast + diploid mesophyll-derived protoplast”. Protoplasts were isolated from embryogenic calli of ‘Pera’ and ‘Westin’ sweet orange cultivars (Citrus sinensis) and from young leaves of ‘Fremont’, Nules’, and ‘Thomas’ mandarins (C. reticulata), and ‘Nova’ tangelo [C. reticulata × (C. paradisi × C. reticulata)]. The regenerated plants were characterized based on their leaf morphology (thickness), ploidy level, and simple sequence repeat (SSR) molecular markers. Plants were successfully generated only when ‘Pera’ sweet orange was used as the embryogenic parent. Fifteen plants were regenerated being 7 tetraploid and 8 diploid. Based on SSR molecular markers analyses all 7 tetraploid regenerated plants revealed to be allotetraploids (somatic hybrids), including 2 from the combination of ‘Pera’ sweet orange + ‘Fremont’ mandarin, 3 ‘Pera’ sweet orange + ‘Nules’ mandarin, and 2 ‘Pera’ sweet orange + ‘Nova’ tangelo, and all the diploid regenerated plants showed the ‘Pera’ sweet orange marker profile. Somatic hybrids were inoculated with Alternaria alternata and no disease symptoms were detected 96 h post-inoculation. This hybrid material has the potential to be used as a tetraploid parent in interploid crosses for citrus scion breeding.

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Acknowledgments

The authors acknowledge the Fundo de Defesa da Citricultura (FUNDECITRUS) for financial support, Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP) for fellowship to LS, and Conselho Nacional de Desenvolvimento Cientifico e Tecnológico (CNPq) for research fellowship to FAAMF, LEAC, MCY, and BMJM.

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Authors and Affiliations

  1. Universidade de São Paulo, Piracicaba, SP, Brazil

    Leonardo Soriano, Francisco de Assis Alves Mourão Filho, Luis Eduardo Aranha Camargo & Beatriz Madalena Januzzi Mendes

  2. Centro APTA Citros Sylvio Moreira, Cordeirópolis, SP, Brazil

    Mariangela Cristofani-Yaly, Rodrigo Rocha Latado, Camila de Andrade Pacheco & Fernando Alves de Azevedo

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  1. Leonardo Soriano
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  2. Francisco de Assis Alves Mourão Filho
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  3. Luis Eduardo Aranha Camargo
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  4. Mariangela Cristofani-Yaly
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  5. Rodrigo Rocha Latado
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  6. Camila de Andrade Pacheco
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  7. Fernando Alves de Azevedo
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  8. Beatriz Madalena Januzzi Mendes
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Correspondence to Beatriz Madalena Januzzi Mendes.

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Soriano, L., de Assis Alves Mourão Filho, F., Camargo, L.E.A. et al. Regeneration and characterization of somatic hybrids combining sweet orange and mandarin/mandarin hybrid cultivars for citrus scion improvement. Plant Cell Tiss Organ Cult 111, 385–392 (2012). https://doi.org/10.1007/s11240-012-0204-z

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  • DOI: https://doi.org/10.1007/s11240-012-0204-z

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