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Submitted to: Plant Cell Tissue and Organ Culture
Publication Type: Peer Reviewed Journal Publication Acceptance Date: 10/21/2005 Publication Date: N/A Citation: N/A Interpretive Summary: We previously discovered that culturing sweet orange callus cells onto semi-permeable membranes composed of cellulose acetate results in the cells forming normal embryos that can be germinated into plants. In this study I isolated protoplasts from the sweet orange callus cells, allowed the protoplasts to divide and form small clusters of cells, cultured those small clusters onto the cellulose acetate membranes, and then determined if embryos were produced that could be germinated into plants. Embryos did form from these small clusters but the embryos did not germinate into plants. The embryos were much smaller than those that formed from the callus cells. Therefore, to obtain embryos capable of germination it is necessary to culture the small clusters to obtain a mass of callus. Technical 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 or 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 15-30 day old p-calli and no abnormal embryo morphologies were observed. The minimum size of p-calli to form embryos was 77.84 ++m in diameter. Embryos were substantially smaller from p-calli than those produced from embryogenic callus and averaged 200 ++m and >1 mm in diameter, respectively. |
