Journal
PLANT CELL REPORTS
Volume 24, Issue 8, Pages 462-467Publisher
SPRINGER
DOI: 10.1007/s00299-005-0969-1
Keywords
Cinnamomum camphora L.; somatic embryogenesis; plant regeneration; protoplast culture
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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 x 10(6)/ 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 center dot 2H(2)O, and 0.011% (w/v) NaH2PO4 center dot 2H(2)O. 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) GA(3). 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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