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Targeted cybridization in citrus: transfer of Satsuma cytoplasm to seedy cultivars for potential seedlessness

  • Genetic Transformation and Hybridization
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Abstract

CMS (cytoplasmic male sterility) can be controlled by the mitochondrion genome in higher plants, including Satsuma mandarin. Somatic fusion experiments in citrus combining embryogenic callus protoplasts of one parent with leaf protoplasts of a second parent often produce cybrid plants of the leaf parent, a phenomenon occurring most often with interspecific fusion combinations. In an attempt to practically exploit this cybridization phenomenon, we conducted somatic fusion experiments combining embryogenic suspension-derived protoplasts of Satsuma mandarin, Citrus unshiu Marc. cv. Guoqing No. 1 (G1), a male-sterile cultivar, with leaf protoplasts of other seedy types—Hirado Buntan Pink pummelo (HBP) [Citrus grandis (L.) Osbeck], Sunburst mandarin (C. reticulata Blanco), Orie Lee hybrid (C. reticulata cv. Clementine × Murcott tangor), and Murcott tangor [C. reticulata × C. sinensis (L.) Osbeck], respectively—in an attempt to generate seedless cybrids by the targeted transfer of CMS. The genetic identities of regenerated plants from all four parental combinations were determined by flow cytometry, SSR, CAPS (or PCR-RFLP), RFLP, and chloroplast-SSR analyses. Regenerated plants from the first three parental combinations were diploids, and the cybrid nature of G1 + HBP with the mitochondrion genome from G1 and the chloroplast genome from HBP was confirmed, whereas the cybrid nature of the remaining two combinations was difficult to confirm because of the close phylogenetic relatedness of both fusion parents, as expected. Plants from G1 + Murcott were confirmed as tetraploid somatic hybrids. This is the first report of targeted citrus cybrid production by symmetric fusion with male-sterile Satsuma as the callus parent and other seedy cultivars as the leaf parents.

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Abbreviations

CAPS::

Cleaved amplified polymorphic sequence

CMS::

Cytoplasmic male sterility

cp-SSR::

Chloroplast simple sequence repeat

PEG::

Polyethylene glycol

SSR::

Simple sequence repeat

RFLP::

Restriction fragment length polymorphism

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Acknowledgements

This research was financially supported by the Florida Agricultural Experiment Station and approved for publication as Journal Series No. R-09483. The authors also thank the Florida Citrus Production Research Advisory Council, the 863 High-Tech Project of China, the National Natural Science Foundation of China (NSFC), the International Foundation for Science (IFS) in Stockholm, Sweden, and the Chengguang Youth Project of Wuhan City of China for partial financial support.

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Authors and Affiliations

  1. National Key Laboratory of Crop Genetic Improvement, National Center of Crop Molecular Breeding, Huazhong Agricultural University, 430070, Wuhan, P.R. China

    W. W. Guo, Y. J. Cheng & X. X. Deng

  2. Department of Biological Sciences, Chicago State University, Chicago, IL, 60628-1598, USA

    D. Prasad

  3. Horticultural Sciences, University of Florida, IFAS, Citrus Research and Education Center, 700 Experiment Station Road, Lake Alfred, FL, 33850, USA

    P. Serrano & J. W. Grosser

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  1. W. W. Guo
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  2. D. Prasad
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  3. Y. J. Cheng
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  4. P. Serrano
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  5. X. X. Deng
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  6. J. W. Grosser
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Correspondence to J. W. Grosser.

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Communicated by G.C. Phillips

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Guo, W.W., Prasad, D., Cheng, Y.J. et al. Targeted cybridization in citrus: transfer of Satsuma cytoplasm to seedy cultivars for potential seedlessness. Plant Cell Rep 22, 752–758 (2004). https://doi.org/10.1007/s00299-003-0747-x

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  • DOI: https://doi.org/10.1007/s00299-003-0747-x

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