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Protoplast isolation and regeneration from the potential economic brown alga Petalonia fascia (Ectocarpales, Phaeophyceae)

Abstract

Petalonia fascia is a widespread brown alga with economic potential due to its use as raw or dried powder, and in the biomedical field. Protoplasts are living plant cells devoid of cell wall with a wide range of applications in basic and applied research, especially in crop improvement, and as a seedstock in seaweeds. Protoplasts have been previously isolated from P. fascia, but, their regeneration ability, an important prerequisite for protoplast applications, has not been explored. In this work, we report the protoplast isolation and successful regeneration from P. fascia using the commercially available cellulase “Onozuka” RS (1%) and alginate lyase (4 U mL−1). Protoplast production was enhanced under increased osmolarity (2512 mOsm L−1 H2O), with chelation pre-treatment, and short incubation time (4 h). Our protocol produced more than 14 times the number of protoplasts obtained using previously reported protocols. After 4 weeks in culture, protoplasts developed into prostrate and discoid and mixed thallus, as well as cell clumps. Blades mostly emerged from prostrate thalli. Antibiotics were not crucial for improving protoplast regeneration, and temperature did not affect the development of the morphological forms and blades. Our results show that high yields of protoplasts (107 protoplasts g−1 FW) with good regeneration ability can be obtained from P. fascia using a simple mixture of commercial enzymes. This represents the first report of successful protoplast regeneration in P. fascia.

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Data availability

The data that support the findings of this study are available from the corresponding author, Tae Oh Cho, upon reasonable request.

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Funding

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2019R1F1A1060346 and NRF-2021R1I1A2059577), by the Ministry of Ocean and Fisheris (MarineBiotics Project, 20210469), and by the National Marine Biodiversity Institute of Korea (2021M01100) to Tae Oh Cho. This research was also supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2021R1I1A1A01051909) to Boo Yeon Won.

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All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Jose Avila-Peltroche. The first draft of the manuscript was written by Jose Avila-Peltroche and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Tae Oh Cho.

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Avila-Peltroche, J., Won, B.Y. & Cho, T.O. Protoplast isolation and regeneration from the potential economic brown alga Petalonia fascia (Ectocarpales, Phaeophyceae). J Appl Phycol 34, 543–550 (2022). https://doi.org/10.1007/s10811-021-02648-0

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Keywords

  • Brown algae
  • Commercial enzymes
  • Petalonia fascia
  • Protoplast isolation
  • Protoplast regeneration