Protoplast production fromPorphyra linearis using a simplified agarase procedure capable of commercial application

Chen, L. C.-M; Craigie, J. S.; Xie, Zhengqing

doi:10.1007/bf02185902

Cited by 27 publications

(8 citation statements)

References 10 publications

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“…Commercial species, such as Porphyra , have been the focus of efforts to obtain propagules from protoplasts. In Porphyra linearis, protoplasts were produced and used as substitutes for conchospores in seeding nets (Chen et al . 1994).…”

Section: Free Cells and Protoplastsmentioning

confidence: 99%

Review: Seaweed tissue culture as applied to biotechnology: Problems, achievements and prospects

Baweja

García‐Jiménez

Robaina

2009

Phycological Research

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Advances have been made in cell and tissue culture of seaweeds to define a unique branch of in vitro techniques; however, they are lagging far behind those of land plants and have limited applications. Explants can be cultivated axenically in enriched or artificial seawater culture media, and regeneration and even callus formation are achieved. In this state of the art technique, seaweed tissue culture may be already useful for certain biotechnological applications, such as clonal propagation of seed material for mariculture. Nevertheless, the absolute control of growth and development as it is exerted in higher plant tissue culture is lacking, and it is required for more complex biotechnological applications in seaweeds. Definitively, we need appropriate cells (competent cells) to induce growth with the most effective chemical regulators in culture medium adjusted towards the addition of carbon sources. Still, free cells and protoplast isolation and regeneration in marine seaweeds constitute the most developed topic in seaweed tissue culture. The regulation of growth and development of seaweed free cell and protoplast cultures may sustain a purposeful use of techniques in the era of genomic applications.

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Section: Free Cells and Protoplastsmentioning

confidence: 99%

Review: Seaweed tissue culture as applied to biotechnology: Problems, achievements and prospects

Baweja

García‐Jiménez

Robaina

2009

Phycological Research

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“…Of the total seaweed protoplast studies reported so far, Porphyra probably is the most studied taxon mainly due to its economic importance. Many of these studies employed enzyme mixtures that contained crude extracts prepared from either marine bacteria or marine herbivorous animal sources for producing protoplasts (Tang, 1982;Polne-Fuller & Gibor, 1984;Saga & Sakai, 1984;Saga et al, 1986;Chen, 1987;Araki et al, 1987;Chen et al, 1988;Yamaguchi et al, 1989;Fujita & Saito, 1990;Waaland et al, 1990;Chen et al, 1994;Packer, 1994;Kito et al, 1998). The usefulness of such enzyme preparations is subject to the availability of such organisms, which are generally confined to specific geographical or climatic conditions.…”

Section: Introductionmentioning

confidence: 98%

Production and seeding of protoplasts of Porphyra okhaensis (Bangiales, Rhodophyta) in laboratory culture

Reddy

Jha

2005

J Appl Phycol

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High yields of viable protoplasts were produced from Porphyra okhaensis H. Joshi, Oza & Tewari following two-step enzymatic digestion (protease pretreatment and cell wall polysaccharides-degrading enzyme treatment) of the thallus. Pretreatment of the tissues with 1% Protease P6 at 20 ± 1 • C for 30 min prior to digestion with cell wall polysaccharide-degrading enzymes increased the protoplast yield two fold compared to tissues that were digested with polysaccharide-degrading enzyme mixture. The polysaccharide-degrading enzymes employed for protoplast isolation from P. okhaensis were Cellulase Onozuka R-10, Macerozyme R-10, abalone acetone powder and agarase. Suitable pH, temperature and duration of enzyme treatment for optimal production of viable protoplasts were pH 6, 20 ± 1 • C and 3 h, respectively. Mannitol (0.8 M) was found to be an excellent osmotic stabilizer. When the tissue of P. okhaensis pretreated with 1% protease solution was digested with commercial enzyme mixture consisting of 2% Cellulase Onozuka R-10, 2% Macerozyme R-10, 1% abalone acetone powder, 50 units of agarase and 0.8 M mannitol in 1% NaCl (adjusted to pH 6.0 with 25 mM MES buffer) with gentle agitation for 3 h at 20 ± 1 • C, 23.2 ± 0.24 × 10 6 protoplasts g −1 fresh wt. were obtained. The regeneration rate of protoplasts isolated in the present study was found to be 79%. Protoplasts that regenerated cell walls underwent regular cell divisions and developed into leafy gametophytic thallus in the laboratory cultures. Further, the seeding of nylon threads with partially developed protoplasts of P. okhaensis was successful in the laboratory conditions and germlings as long as 3-4 cm were obtained from such seeded threads in one month period in aerated cultures.

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“…Since the 1970s, many researches have been devoted to developing a highly efficient monospore-release method to propagate seedlings. Researchers have used cellulase, pectinase and enzymes from the digestive gland of shellfish or mixtures of these enzymes to isolate protoplasts that could then develop into monospores (Polne-Fuller and Gibor 1984;Waaland et al 1990;Chen et al 1994). However, protoplasts are sensitive to enzymatic digestion.…”

Section: Discussionmentioning

confidence: 99%

Thallus sectioning as an efficient monospore release method in Pyropia yezoensis (Bangiales, Rhodophyta)

et al. 2019

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The red seaweed Pyropia yezoensis is a marine crop of important economic value and is widely cultivated in the coastal areas of northern China. The current seedling-raising system relies on the germination of conchospores, a process not fast enough to meet the increasing demand from farmers. In this study, we developed a monospore-dependent seedling method based on the asexual reproduction of the Pyropia thallus. The Pyropia thallus was physically sectioned into small pieces (microthalli) that were cultivated at 15°C. The algal cells in the microthalli became morphologically condensed, underwent cell division, and then developed into monospores on the 5th day. The monospores were able to attach to the seeding rope in 24 h and germinated into healthy thalli. To optimize the efficiency of monospore release, we tested the effect of temperature and the size and original positions of the microthalli as well as the age of the mother thallus. Microthalli with a size of 30-50 cells from the middle and apex of 21-day-old thalli cultivated at 15°C yielded the optimum production of monospores. Theoretically, in this thallus-tothallus seeding strategy, 0.1 g of thallus could produce at least 10 7 monospores, with the same number of offspring thallus seedlings. Taking all of these advantages together, including the high efficiency, short time, low cost and easy operation, this physical sectioning method could serve as a promising seed source especially for new cultivars with superior traits in Pyropia farming and rapid replenishment of seedlings when thalli undergo disastrous diseases.

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Protoplast production fromPorphyra linearis using a simplified agarase procedure capable of commercial application

Cited by 27 publications

References 10 publications

Review: Seaweed tissue culture as applied to biotechnology: Problems, achievements and prospects

Review: Seaweed tissue culture as applied to biotechnology: Problems, achievements and prospects

Production and seeding of protoplasts of Porphyra okhaensis (Bangiales, Rhodophyta) in laboratory culture

Thallus sectioning as an efficient monospore release method in Pyropia yezoensis (Bangiales, Rhodophyta)

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