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In vitro production of autotetraploid Ponkan mandarin (Citrus reticulata Blanco) using cell suspension cultures

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Abstract

An efficient protocol for colchicine mediated production of in vitro autotetetraploids from Ponkan mandarin using cell suspension cultures is described. Cells were treated with 1 g l−1 colchicine for 4 or 8 days before transfer into solid EME medium supplemented with 5% maltose. Colchicine treated cells were placed in medium with or without an overlay of 1:2 medium–mixture of liquid 0.6 M BH3 medium and 0.15 M EME + maltose liquid media. It was observed that modifying the immediate cell environment by addition of the liquid overlay played a positive role in cell differentiation and subsequent plant regeneration. Ploidy levels were determined with a flow cytometer and confirmed by chromosome staining using the enzymatic maceration method. A large number of non-chimeric autotetraploids were generated using this method. Such plants have great value in a breeding program for the development of seedless triploid citrus, as very few available tetraploid breeding parents are easy to peel.

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Abbreviations

DAPI:

4′,6-Diamidino-2-phenylindole

MT:

Murashige and Tucker

UV:

Ultraviolet

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Authors and Affiliations

  1. Horticultural Sciences Department, Citrus Research and Education Center, University of Florida-IFAS, 700 Experiment Station Road, Lake Alfred, Fl, 33850, USA

    M. Dutt, M. Vasconcellos, K. J. Song, F. G. Gmitter Jr. & J. W. Grosser

  2. Faculty of Bioscience and Industry, Cheju National University, 66 Jejudaehakno, Jeju, 690-756, Korea

    K. J. Song

Authors
  1. M. Dutt
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  2. M. Vasconcellos
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  3. K. J. Song
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  4. F. G. Gmitter Jr.
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  5. J. W. Grosser
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Correspondence to J. W. Grosser.

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Dutt, M., Vasconcellos, M., Song, K.J. et al. In vitro production of autotetraploid Ponkan mandarin (Citrus reticulata Blanco) using cell suspension cultures. Euphytica 173, 235–242 (2010). https://doi.org/10.1007/s10681-009-0098-y

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  • DOI: https://doi.org/10.1007/s10681-009-0098-y

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