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Isolation and callus formation of Gracilariopsis bailiniae (Gracilariales, Rhodophyta) protoplasts

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Abstract

This paper reports the first successful isolation of protoplasts from Gracilariopsis bailiniae and their callus formation. The base solution type, concentration of isolating enzymes, concentration of sorbitol, incubation time, temperature and pH of the enzyme solution were tested to optimize the protoplast yield. The optimized isolation conditions were: 40% base solution 3 (deionized water containing 25 mmol/L MESTris and 25 mmol/L CaCl2·2H2O) and 60% crude Marinomonas sp. YS-70 agarase solution, containing 2% w/v cellulase, 1% w/v macerozyme R-10 and 0.4 mol/L sorbitol, with incubation for 4 h at 28°C and pH 6.5. The highest yield of viable protoplasts, which was obtained in these conditions, was (1.75±0.25)×10 6 cells/g fresh weight. Cell wall regeneration of most protoplasts from G. bailiniae was complete within 60 h and the first division of cells happened after ≥3 days. Two division types were observed in the first division of protoplasts from G. bailiniae— asymmetric division and symmetric division. After the first division, the cells underwent a series of divisions to form callus cell masses.

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Acknowledgement

The authors thank Professor LIU Tao (Ocean University of China), Professor MEI Zhiping (Shantou University), Dr. WANG Hui (Shantou University) and Ms. WANG Zhongxia (Ocean University of China) for providing much advice and kind assistance.

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Authors and Affiliations

  1. Marine Biology Institution, Key Laboratory of Marine Biotechnology of Guangdong Province, Shantou University, Shantou, 515063, China

    Haihong Chen  (陈海红), Weizhou Chen  (陈伟洲), Jingyi Shi  (石经仪), Zepan Chen  (陈泽攀) & Yi Zhang  (张毅)

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  1. Haihong Chen  (陈海红)
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  2. Weizhou Chen  (陈伟洲)
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  3. Jingyi Shi  (石经仪)
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Correspondence to Weizhou Chen  (陈伟洲).

Additional information

Supported by the China Agriculture Research System (No. CARS-50), the Science and Technology Program of Guangdong Province of China (Nos. 2016A020222023, 2015B090903081), and the Project of Guangdong Province Education Department (No. 2017KCXTD014)

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Chen, H., Chen, W., Shi, J. et al. Isolation and callus formation of Gracilariopsis bailiniae (Gracilariales, Rhodophyta) protoplasts. J. Ocean. Limnol. 36, 2268–2277 (2018). https://doi.org/10.1007/s00343-019-7217-y

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  • DOI: https://doi.org/10.1007/s00343-019-7217-y

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