Optimization of protoplast isolation from the gametophytes of brown alga Undaria pinnatifida using response surface methodology

Avila-Peltroche, José; Won, Boo Yeon; Cho, Tae Oh

doi:10.1007/s10811-020-02095-3

Cited by 7 publications

(2 citation statements)

References 51 publications

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“…Our results in Scytosiphon lomentaria and Dictyopteris pacifica suggest that only two components, cellulase and alginate lyase, are sufficient for protoplast production in brown algae. This is consistent with the findings of previous works on filamentous brown algae [ 11 , 12 , 45 ].…”

Section: Discussionsupporting

confidence: 94%

Protoplast isolation from Dictyopteris pacifica and Scytosiphon lomentaria, using a simple commercial enzyme preparation

Avila-Peltroche

Won

Cho

2021

Journal of Genetic Engineering and Biotechnology

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Background Protoplasts (i.e., naked plant cells) can be used for in vitro manipulations and genetic improvement in cultivars with economic value. During the last decade, protoplast research in economic brown algae has been scarce, and it is usually hampered by the use of non-commercial enzymes or crude extracts for isolating protoplasts. Dictyopteris pacifica is part of a brown algal genus well known by its wide chemical diversity and biological properties. Scytosiphon lomentaria is an edible brown seaweed with antioxidant, antitumor, and antiviral properties. So far, there are no protoplast isolation protocols using commercial enzymes for these two economic brown algae. In this study, we obtained protoplasts from cultured samples of D. pacifica and S. lomentaria using commercially available enzymes. Additionally, we investigated the effects of Driselase inclusion and Ca-chelation pre-treatment on protoplast yields in order to optimize the conditions for protoplast preparations. Results Protoplasts were isolated from Dictyopteris pacifica and Scytosiphon lomentaria using the commercially available Cellulase Onozuka RS (1%) and Alginate lyase (4 U mL−1), and short incubation time (4 h). Driselase did not show significant effects on protoplast production in both species. Ca-chelation pre-treatment only increased the number of protoplasts in D. pacifica. Under optimal conditions, the protoplast yields from D. pacifica and S. lomentaria were 4.83 ± 2.08 and 74.64 ± 32.49 × 106 protoplasts g−1 fresh weight, respectively. The values obtained for S. lomentaria were 2–3 orders of magnitude higher than previously reported. Conclusions Our results show that high protoplast yields can be obtained from D. pacifica and S. lomentaria using a simple mixture of commercial enzymes (Cellulase RS and Alginate lyase) and short incubation time (4 h). This work also represents the first report of protoplast isolation in D. pacifica. The method proposed here can help to expand protoplast technology in more brown algal species.

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Section: Discussionsupporting

confidence: 94%

Protoplast isolation from Dictyopteris pacifica and Scytosiphon lomentaria, using a simple commercial enzyme preparation

Avila-Peltroche

Won

Cho

2021

Journal of Genetic Engineering and Biotechnology

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“…For example, Rhodophyta only has the classes Bangiophyceae [ 13 , 14 , 15 ], Florideophyceae [ 13 ], and Porphyridiophyceae [ 14 ]. Most studied brown algae are from the same class Phaeophyceae, e.g., Dictyopteris pacifica, Scytosiphon lomentaria [ 15 ], Petalonia fascia [ 9 ], and Undaria pinnatifida [ 16 ]. In contrast, there are more green algae that have been studied recently, such as Chlorella protothecoides [ 17 ], Ulva prolifera [ 18 ], Haematococcus pluvialis [ 19 ], and Penium margaritaceum [ 20 ].…”

Section: Introductionmentioning

confidence: 99%

Protoplast Preparation for Algal Single-Cell Omics Sequencing

Yang

Xia

et al. 2023

Microorganisms

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Single-cell sequencing (SCS) is an evolutionary technique for conducting life science research, providing the highest genome-sale throughput and single-cell resolution and unprecedented capabilities in addressing mechanistic and operational questions. Unfortunately, the current SCS pipeline cannot be directly applied to algal research as algal cells have cell walls, which makes RNA extraction hard for the current SCS platforms. Fortunately, effective methods are available for producing algal protoplasts (cells without cell walls), which can be directly fed into current SCS pipelines. In this review, we first summarize the cell wall structure and chemical composition of algal cell walls, particularly in Chlorophyta, then summarize the advances made in preparing algal protoplasts using physical, chemical, and biological methods, followed by specific cases of algal protoplast production in some commonly used eukaryotic algae. This review provides a timely primer to those interested in applying SCS in eukaryotic algal research.

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

2022

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Optimization of protoplast isolation from the gametophytes of brown alga Undaria pinnatifida using response surface methodology

Cited by 7 publications

References 51 publications

Protoplast isolation from Dictyopteris pacifica and Scytosiphon lomentaria, using a simple commercial enzyme preparation

Protoplast isolation from Dictyopteris pacifica and Scytosiphon lomentaria, using a simple commercial enzyme preparation

Protoplast Preparation for Algal Single-Cell Omics Sequencing

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

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