Isolation and Culture of Mesophyll Protoplasts from In vitro Subcultured Japanese White Birch

概要

論文の詳細を見る
成木の葉柄培養によって得されたシラカンバ幼植物体の葉の組織から，酵素液を用いてプロトプラストを単離することができた。プロトプラストの収量は葉1gあたり約6×106個であった。単離されたプロトプラストを2，4－Dを3mg/l含む，一部改変したMS液体培地に2×106個の密度で培養した。その結果，確かな細胞分裂は観察されなかったが，培養3週間後には活発なバディング現象が観察された。Mesophyll protoplasts were successfully isolated from leaf tissue of in vitro cultured Japanese white birch (Betula platyphylla var. japonica) plantlets which were originated from shoot primordia developed on petiole culture of mature tree. Approximately 6×106 protoplasts were isolated from about 1g of leaf tissue by enzyme solution. The protoplasts were cultured in liquid modified MS (MURASHIGE and SKOOG, 1962) medium with 3mg/l of 2,4-D(2,4-dichlorophenoxyacetic acid) at the density of about 2×106. Although the certain cell divisions were not observed, active cell buddings were observed after three weeks of protoplasts culture.