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Regeneration of somatic embryos from sweet orange (C. sinensis) protoplasts using semi-permeable membranes

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Abstract

Sweet orange (C. sinensis L. Osbeck) protoplasts were isolated from nucellar-derived embryogenic callus, cultured in alginate beads for 5–30 days, and the resulting p-calli released by liquefaction and cultured on semi-permeable membranes overlaid on MT culture medium. Somatic embryos did not develop from 5- to 10-day-old p-calli but did develop from 15-, 20-, 25-, and 30-day-old p-calli. There were no significant differences in the numbers of embryos produced among the 15- to 30-day-old p-calli and no abnormal embryo morphologies were observed. The minimum size of p-calli to form embryos was 77.84 μm in diameter. Embryos were smaller from p-calli than those produced from embryogenic callus; p-calli-derived embryos ranged in size between 0.5 and 0.8 mm, while embryos derived from embryogenic callus ranged between 1 and 2 mm.

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Abbreviations

BA:

6-benzylaminopurine

CPM1:

citrus protoplast medium 1

GA:

gibberellic acid

HS:

heart-shaped

MC:

multiple cotyledon

MT:

Murashige and Tucker

p-calli:

protoplast-derived calli

PE:

plating efficiency

SE:

standard error

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Acknowledgement

I wish to thank Mr Eldridge Wynn for his excellent assistance with these experiments.

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Authors and Affiliations

  1. Agricultural Research Service, US Horticultural Research Laboratory, 2001 South Rock Road, Ft. Pierce, FL, 34945-3030, USA

    Randall P. Niedz

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  1. Randall P. Niedz
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Correspondence to Randall P. Niedz.

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Niedz, R.P. Regeneration of somatic embryos from sweet orange (C. sinensis) protoplasts using semi-permeable membranes. Plant Cell Tiss Organ Cult 84, 353–357 (2006). https://doi.org/10.1007/s11240-005-9028-4

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  • DOI: https://doi.org/10.1007/s11240-005-9028-4

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