Isolation of protoplasts from fern prothallia and their regeneration to gametophytes
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Protoplasts were isolated enzymatically from prothallia ofLygodium japonicum. The protoplasts grown in a culture medium containing 0.6 M mannitol and 0.05 M sucrose began to divide within 8 days of culture, and after 30 days 10-cell clusters were present. When the cell-clusters were transferred into fresh media followed by sequential reduction of mannitol concentration, they developed rhizoids and protonemata. The reduction of mannitol concentration to 0.3 M resulted in the regeneration of a common gametophyte within 50 days of culture, and subsequently the regenerated gametophytes produced sporophytic leaves and roots.
Key wordsFern gametophyte Lygodium Prothallium Protoplast Regeneration
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