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In vitro anther culture of sweet orange (Citrus sinensis L. Osbeck) genotypes and of a C. clementina × C. sinensis ‘Hamlin’ hybrid

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Abstract

Citrus, and particularly sweet oranges, are very recalcitrant to anther culture. In this paper it was evaluated for the first time the response of 27 genotypes of Citrus sinensis and of one hybrid C. clementina × C. sinensis, to in vitro anther culture. Ten genotypes of sweet oranges showed embryogenic callus induction, mostly blood sweet oranges genotypes, such as Tarocco, Moro and Sanguinelli. In vitro microspore developmental switches from the gamethophytic to the sporophytic pathway were shown by DAPI staining in microspores of these responsive genotypes, after 10 months in culture. However, microsatellite marker analyses showed that these calli were heterozygous. The flow-cytometric analysis of these embryogenic calli showed the presence of two peaks, corresponding to haploid (n) and diploid (2n) genotypes. Differently, anther cultures of the hybrid C. clementina × C. sinensis produced tri-haploid (3n) embryogenic calli and the embryos obtained were homozygous when analyzed by molecular markers (sample sequence repeats), confirming the more responsive characteristic of clementine to microspore embryogenesis through anther culture.

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Acknowledgments

JCC and RRL thank to FAPESP for doctoral scholarship (2008/09928-8) and for financial support of the project (2008/10.203-8), and APM acknowledges CNPq (310.612/2011-0) for the research fellowship. The authors also thank the biologist Mônica Lanzoni Rossi (CENA/USP) for assistance processing the samples for anther microscopic analysis (Fig. 2b) presented in this manuscript.

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  1. Jean C. Cardoso

    Present address: Departamento de Desenvolvimento Rural, CCA/UFSCar, Rod. Anhanguera, km 174, Araras, SP, Brazil

Authors and Affiliations

  1. University of São Paulo, CENA, Av. Centenário, 303, Piracicaba, SP, 13416-903, Brazil

    Jean C. Cardoso & Adriana P. Martinelli

  2. Dipartimento di Scienze Agrarie e Forestali, Università degli Studi di Palermo, Viale delle Scienze, 11, 90128, Palermo, Italy

    Maria Antonieta Germanà

  3. Centro APTA Citros Sylvio Moreira/IAC, Rod. Anhanguera, km 187, Cordeirópolis, SP, Brazil

    Rodrigo R. Latado

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  1. Jean C. Cardoso
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Cardoso, J.C., Martinelli, A.P., Germanà, M.A. et al. In vitro anther culture of sweet orange (Citrus sinensis L. Osbeck) genotypes and of a C. clementina × C. sinensis ‘Hamlin’ hybrid. Plant Cell Tiss Organ Cult 117, 455–464 (2014). https://doi.org/10.1007/s11240-014-0456-x

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