【摘要】 以我校选育的94591、373和953863个油菜品种的花粉植株为材料，切取其无菌试管苗幼嫩叶片，将其切成1～2mm细条游离原生质体。原生质体培养采用液体浅层培养、双层培养法。培养基为加有不同细胞分裂素、生长素和天然有机成分的MS液体培养液。原生质体在培养3～6d开始第一次分裂，分裂频率最高可达14．0％，20d后形成多细胞团，35～40d即可形成肉眼可见的愈伤组织。将愈伤组织增殖后转移到附加了细胞分裂素6－BA和GA3的MS固体培养基上进行分化培养，40～60d后，品种94591的愈伤组织分化出不定芽。另二个品种的愈伤组织逐步褐化，未见分化。试管苗转入添加IAA和NAA的生根培养基培养获得了再生植株。更多还原

【Abstract】 Protoplasts were isolated enzymatically from leaf blades of pollen plant ofrapeseed (Brassica napus L.) cultivars 94591,373 and 95386. The purified protoplasts were cultured with two different methods on MS medium supplemented with cytokinin, auxin and other natural organic compositions. The method called "double layer culture" (including thin agarose solid layer and thin liquid layer) was found superior to the method of thin liquid culture. Protoplasts obtained the first division after 4-6 days in culture. The division frequency reached 14.0% and microlonies were formed after 20 days. Microcalli were formed after 35-40 days in culture. Calli were transfered to proliferation and regenef’ation media with 6-BA and GA3 for 40-60 days in cultore, the calli of rapeseed 94591 formed tube-shoots, orther two variety calli did not regenerate any tube-shoots.Plats regenerated from shoots when they were cultured on MS medium with IAA and NAA.更多还原