摘要
从室温(20～25℃)和自然光照下生长的青花菜(Brassica oleracea var.italica)无菌苗下胚轴游离的原生质体,纯化后的得率为1×10~6/g fw。采用琼脂糖小块培养(Agarose Bead Cul-ture)方法,培养在 K8P 补加 NAA 1.0mg/L、2,4-D 0.1 mg/L、ZT 0.5 mg/L 和琼脂糖0.6%培养基中的原生质体,获得了持续的细胞分裂,并形成了小愈伤组织,分裂频率可达35%,植板率为5%。这表明无菌苗的条件和琼脂糖小块培养方法是适用于青花菜下胚轴原生质体游离和培养的。小愈伤组织增殖后或直接转入 MS 补加 IAA 0.1 mg/L、ZT 2.0 mg/L、Adenine 40 mg/L 和 Glu-tamine 100 mg/L 的分化培养基中能分化出大量新梢,分化率可达20%。切下新梢后移入 MS 补加IBA 0.5 mg/L 的生根培养基中,易生根形成完整植株,并易移栽成活。
Hypocotyl protoplasts were isolated from sterile seedlings of broccoli(Brassi- ca oleracea var.italica)cultured at room temperature(20～25℃)and natural illumination.Af- ter the purification of protoplasts,higher protoplast yield (1×10~6/g·fw) was obtained.Us- ing agarose bead culture technique,the purified protoplasts were cultured in K8P medium supplemented with NAA 1.0mg/L,2,4-D 0.1mg/L,ZT 1.0 mg/L and agarose 0.6%.The continued division of regenerated cells and formation of small calli were observed,and the di- vision frequency and plating efficiency were about 35% and 5% ,respectively.It indicated that the condition of sterile seedlings and agarose bead culture technique are suitable for the isola- tion and culture of hypocotyl protoplasts of broccoli.Multiple shoots were induced on differ- entiation medium of MS supplemented with IAA 0.1mg/L,ZT 2.0 mg/L,adenine 40 mg/L and glutamine 100 mg/L,and the differentiating rate reached 20%.After transferring differ- entiated shoots onto a rooting medium of MS supplemented with IBA 0.5 mg/L,the shoots rooted and the plantlets were transplanted into soil and survived easily.
出处
《上海农业学报》
CSCD
1993年第4期13-18,共6页
Acta Agriculturae Shanghai
关键词
青花菜
下胚轴
原生质体
再生植株
Broccoli
Hypocotyl
Protoplast
Regenerated plantlet
