摘要
山杉木(Cunninghamia lanccolata Hook.)成熟种子胚在含有2.0mg/l 2,4-D、0.2mg/l KT的MS培养基上诱导愈伤组织,通过筛选继代培养,得到了生长旺盛、质地松散的愈伤组织细胞系。继代初期适当降低蔗糖浓度有助于减少愈伤组织的褐化。将5个月龄的愈伤组织转移至含1.5~2.0 mg/l 2,4-D、0.1~0.2mg/l KT、200 mg/l CH的MS液体培养基振荡培养,建立了增殖快、分散性良好的杉木悬浮细胞系。从4个月龄的悬浮细胞酶解(2%纤维素酶、0.25%果胶酶和13%甘露醇),游离得到大量的杉木原生质体,产量为9.3×10~5个/g FWW,活力达到80%以上。
Callus was induced by embryos from mature seeds of Chinese fir (Cttnninghamia lanceolata Hook.), which had been cultured in MS medium containing 2.0mg/l 2,4-D and 0.2mg/l KT. After successive subculture and selection, cell colonies of the calli which were frible and in rapid growth were obtained. In the period of initial subculture, decrease of sucrose concentration was beneficial to diminishing callus browning. The cell suspension culture, which grew rapidly and consisted mainly of smaller cell aggregates, was established by transferring five-month-old calli to liquid MS medium supplemented with 1.5~2.0mg/l 2.4-D, 0.1~0.2mg/l KT and 200 mg/1 CH. A great amount of Chinese Fir protoplasts were released from the cell suspension cultures, with protoplast yield (9.3x105/g FW) and viability (above 80 %). Enzyme mixture was consisted of 2 % cellulase R-10, 0.25% pectinase and 13% mannitol.
出处
《林业科学研究》
CSCD
北大核心
1992年第6期628-632,共5页
Forest Research
基金
中国林科院及亚热带林业研究所科研基金
关键词
杉木
悬浮细胞系
原生质体的分离
Chinese Fir cell suspension culture protoplast isolation