Abstract
Morus alba(white mulberry) mesophyll protoplasts were isolated from leaves of 30-45 day old sterile shoots, with protoplast yields of 2.5 × 107 g−1/F.W. after purification. The protoplasts were cultured in a modified K8P liquid medium containing 0.2 mg/L 2,4-D(2,4- Dichlorophenoxy acetic acid), 1 mg/L NAA(Naphthyl acetic acid) and 0.5 mg/L BA(6-benzylaminopurine). A low plating density (5 × 104/ml) proved to be favourable to the division of protoplast-derived cells. The first division occurred 4 days after culture, and the division frequency reached 24% at 10 days. A number of cell colonies and microcalli formed in 6 weeks. The microcalli were transferred onto MSB medium with 0.5 mg/L NAA and 0.5 mg/L BA for further proliferation. Shoot forrgation was initiated when the calli of 3-4 mm in size were transferred onto MSB differentiation medium with 0.1 mg/L NAA and 1 mg/L BA. The frequency of shoot formation was 35%. The shoots of 4-5 cm in height were excised from the callus and rooted on half strength MS medium with 0.5 mg/L IBA and 0.1 mg/L BA. After transplantation into pots, the regenerated plants grew vigorously in the phytotron.
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This research work was supported by the National Biotechnology Programme and the Biotechnology Projects of the Ministry of Forestry (1991-1995).
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Wei, Z., Xu, Z., Huang, J. et al. Plants regenerated from mesophyll protoplasts of white mulberry. Cell Res 4, 183–189 (1994). https://doi.org/10.1038/cr.1994.19
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DOI: https://doi.org/10.1038/cr.1994.19
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