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Studies on the isolation and culture of protoplasts from Kappaphycus alvarezii

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Abstract

In this study, protoplasts were successfully isolated from Kappaphycus alvarezii using snail enzymes, abalone enzymes and cellulase. The optimum enzymic ratio was fixed to be 20% of abalone enzyme, 12% of cellulase and the osmotic stabilizer was 2.0 mol/L glucose. The optimum enzymic hydrolysis conditions were found to be dark enzymolysis at 30°C continuing for 4.0 h. The resultant density and yield of protoplasts achieved 32.60×104 mL−1, 65.20×104 g−1 tissue for Kappaphycus alvarezii. Finally, under the temperature of 20°C, light intensity of 1 500–2 000 lx and photoperiod of 12 h/d, two developmental pathways were investigated: (1) callus-like cell mass and regenerated plantlet occurred on protoplast; (2) young shoots and callus-like cell mass occurred in tissue blocks after enzymolysis.

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Authors and Affiliations

  1. National Marine Environmental Forecasting Center, Beijing, 100081, China

    Si Zhang

  2. College of Marine Life Sciences, Ocean University of China, Qingdao, 266003, China

    Si Zhang, Cui Liu, Yuemei Jin, Shan Chi & Tao Liu

  3. Hainan Provincial Fisheries Research Institute, Haikou, 570204, China

    Xianming Tang & Fuxiao Chen

  4. School of Life Science, Shandong University, Jinan, 250100, China

    Xu Fang

Authors
  1. Si Zhang
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  2. Cui Liu
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  3. Yuemei Jin
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  4. Shan Chi
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  5. Xianming Tang
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  6. Fuxiao Chen
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  7. Xu Fang
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  8. Tao Liu
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Corresponding author

Correspondence to Tao Liu.

Additional information

Foundation item: The National Science Foundation Project under contract No. 2007FY210500; the National Department Public Benefit Research Foundation of China under contract No. 200805075; the Province Science and Technology in the Guangdong Project under contract Nos 2010B060200010 and 2010B020201015; the Science Expenditure in the Hainan Project under contract No. 11-20410-0015; the National Natural Science Foundation of China under contract Nos 41206106 and 41222038.

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Zhang, S., Liu, C., Jin, Y. et al. Studies on the isolation and culture of protoplasts from Kappaphycus alvarezii . Acta Oceanol. Sin. 33, 114–123 (2014). https://doi.org/10.1007/s13131-014-0546-y

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  • DOI: https://doi.org/10.1007/s13131-014-0546-y

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