【摘要】 以黄将甘蓝试管苗幼叶为材料分离原生质体，用ＫＭ８Ｐ培养基进行液体浅层培养和薄层漂浮培养。原生质体的培养密度为６×１０３～１０４／ｍｌ。在薄层漂浮培养中，２天后开始第一次分裂；第５天和７天的分裂频率分别达２３％和３０％。培养１０天后加入稀释培养液，并从暗培养转至弱光照条件下培养，以后每周加一次稀释液。培养１５天后形成小细胞团；３０～３５天形成肉眼可见的小愈伤组织，植板率为３．５％。愈伤组织在含ＩＢＡ０．５～１．０ｍｇ／Ｌ，６—ＢＡ１．５～５．０ｍｇ／Ｌ的分化培养基上，培养约１个月可获得再生无根苗，分化频率可达６８．５％。再生小苗转入合ＮＡＡ０．２ｍｇ／Ｌ的生根培养基，可获得具根的完整再生小植株。更多还原

【Abstract】 Protoplasts of yellow-seed cabbage (Brussica oleracea )which were isolated enzymatically from young leaves of cultured shots,were cultured in KM8P medium with two methods,The density of cultured protoplasts was ranging from 6×103/ml to 1×103ml. Following cell wall regeneration, protoplasts cultured in floating thin layer showed rapid division and high frequency of cell division conpsred with those cultured in liquid. The first division was observed after 2 days in thin floating layer culture. The division frequency reached 23% and 30% at day 5 and day 7,respctively. The initiation of cell division in case of liquid culture was late,usualy at day 5,and the division frequency waw also some what lower. 10 days after culture,a dilution procedure was repeated at one week interval thereafter,and were thansferred to light condition with an intensity of about 800-1000 Lx. The protocolonies were formed after being cultured for 15 days. About one month after culture,microplli were observed with naked eye. A 3. 5% plating efficiency was achieved. Shoots regeneration occurred in one month when callus were transferred onto WS agar media containing indol butyric acid (IBA) 0. 5-1.0mgA, 6-benzylaminopurine6-BA )1. 5-5.0mg/L. The frequency of shoot regeneration was up to 68. 5%. Individual shoots were rooted on a rooting medium supplemented with a naphthalene acetic acid (NAA)0. 2mg/. Intact plantlets with normal morphology were eventuslly transplanted into a Soil mixture.更多还原