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Plant regeneration from protoplasts isolated from embryogenic suspension cultured cells of Cinnamomum camphora L.

  • Cell Biology and Morphogenesis
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Abstract

An efficient and reproducible protocol is described for the regeneration of Cinnamomum camphora protoplasts isolated from cultured embryogenic suspension cells. Maximum protoplast yield (13.1±2.1×106/g FW) and viability (91.8±3.8%) were achieved using a mixture of 3% (w/v) cellulase Onozuka R10 and 3% (w/v) macerozyme Onozuka R10 in 12.7% (w/v) mannitol solution containing 0.12% (w/v) MES, 0.36% (w/v) CaCl2·2H2O, and 0.011% (w/v) NaH2PO4·2H2O. First divisions occurred 7–10 days following culture initiation. The highest division frequency (24.6±2.9%) and plating efficiency (6.88±0.8%) were obtained in liquid medium (MS) supplemented with 30 g l−1 sucrose, 0.7M glucose, 0.1 mg l−1 NAA, 1.0 mg l−1 BA, and 1.0 mg l−1 GA3. After somatic embryo induction and then shoot induction, the protoplast-derived embryos produced plantlets at an efficiency of 17.5%. Somatic embryos developed into well-rooted plants on MS medium supplemented with 1.0 mg l−1 3-indole butyric acid (IBA). Regenerated plants that transferred to soil have normal morphology.

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Abbreviations

BA::

6-Benzyladenine;

CH::

Casein hydrolysate;

CPW::

Cell protoplast wash;

GA3::

Gibberellic acid;

IBA::

3-Indole butyric acid;

NAA::

α-Naphthylacetic acid;

ME::

Malt extract;

MES::

2-(N-morpholino) Ethanesulphonic acid;

MS::

Murashige and Skoog medium;

PGR::

Plant growth regulator;

SAD::

Adenine sulphate

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Correspondence to Manzhu Bao.

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Communicated by R.J. Rose

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Du, L., Bao, M. Plant regeneration from protoplasts isolated from embryogenic suspension cultured cells of Cinnamomum camphora L.. Plant Cell Rep 24, 462–467 (2005). https://doi.org/10.1007/s00299-005-0969-1

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  • DOI: https://doi.org/10.1007/s00299-005-0969-1

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