Plant regeneration from protoplasts isolated from embryogenic suspension cultured cells of Cinnamomum camphora L. |
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Authors: | Li Du Manzhu Bao |
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Institution: | (1) Key Laboratory of Horticultural Plant Biology of Ministry of Education, College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan, 430070, P. R. China |
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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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Keywords: | Cinnamomum camphora L Somatic embryogenesis Plant regeneration Protoplast culture |
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