スサビノリ野生株とアサクサノリ緑色変異株からのプロトプラストの作出と再生

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Porphyranase, β-1, 4-mannanase, and β-1, 3-xylanase, capable of degrading the cell wall of red algae, genus Porphyra, were prepared from the culture fluids of Vibrio sp. AP-2, Aeromonas sp. F-25, and Vibrio sp. AX-4, respectively, The thalli of P. yezoensis, after treatment with a 2% papain solution, were suspended in 5ml of MES buffer, pH 6.0, containing 0.7M mannitol and 1 unit each of porphyranase, β-1, 4-mannanase, and β-1, 3-xylanase. When a 100mg weight of the thalli was shaken in the enzyme solution at 22°C for 90 min, ca. 8.6×105 protoplasts were released.Protoplasts from P. yezoensis grew in a ASP12 (NTA) medium and developed into brown thalli of ca. 10cm in length after 3 months. A number of protoplasts were obtained from green variant of P. tenera, too. These green protoplasts developed into green thalli of ca. 10cm in length after 2 months. A lot of monospores released from regenerated thalli of P. yezoensis (wild type) and P. tenera (green type) developed into brown thalli and green thalli, respectively.
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