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Official Journal of the Japan Wood Research Society

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Regeneration of protoplasts from hyphal strands ofVolvariella volvacea

Journal of Wood Science volume 44, pages 401–407 (1998)Cite this article

Abstract

A series of experiments on the preparation and regeneration of protoplasts from hyphal strands ofVolvariella volvacea (Bull. ex. Fr.) Singer were conducted with the aim of optimizing the conditions for its efficient regeneration. One commercial (Vvcl) and two wild (EAAC-0001 and EAAC-0002) strains ofV. volvacea from the Philippines were used and subjected to varying conditions to determine the most efficient means for regeneration of their protoplasts. The effects of age and type of strain, pH, type and concentration of osmotic stabilizer, enzymatic composition, treatment time, temperature, reciprocal frequency during enzymatic lysis of the cell wall, and centrifugation conditions were investigated. Results showed that the three strains ofV. volvacea had varying responses in terms of yield, size, and ability of their protoplasts to regenerate into the protoplast regeneration medium. Among the three strains, EAAC-0002 had the highest rate of regeneration. The 5-day-old culture ofV. volvacea, when subjected to a combination of 2% Novozyme 234 and 0.2% chitinase in 0.6M mannitol (pH 6.0) for 3 h at 30°C, 90 strokes/min and centrifuged at 1100 g for 10 min; produced an efficient yield of protoplasts with a relatively high regeneration rate.

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Authors and Affiliations

  1. Faculty of Agriculture, Department of Forestry, Tokyo University of Agriculture, 156-8502, Tokyo, Japan

    Renato G. Reyes, Fumio Eguchi, Tomoaki Iijima & Miyato Higaki

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  1. Renato G. Reyes
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  2. Fumio Eguchi
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  3. Tomoaki Iijima
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  4. Miyato Higaki
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Reyes, R.G., Eguchi, F., Iijima, T. et al. Regeneration of protoplasts from hyphal strands ofVolvariella volvacea . J Wood Sci 44, 401–407 (1998). https://doi.org/10.1007/BF01130455

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