摘要
用峨参茎节来源的胚性愈伤组织,在含1.5%纤维素酶Onozuka R-10+0.5%果胶酶Macerozyme R-10+0.5%蜗牛酶+5mmol/L CaCl_2+0.6mol/L甘露醇+0.3%葡聚糖硫酸钾,pH5.8的酶液中游离原生质体,培养在改良MS培养基中,内含激素1mg/L2,4-D+0.5mg/L玉米素,并附加0.3%琼脂糖。第10天分裂频率达40—50%。此后振荡培养1个月,细胞团移至含0.5mg/L 2,4-D及渗透压降低的MS培养基中大量繁殖,然后再移至无激素或含0.1mg/L玉米素的MS培养基中进行分化,得到大量的球形胚,并且同步化达90%,70%的球形胚在10—15天后同步发育为心形胚,以后继续发育为鱼雷胚及子叶胚,最后形成完整植株。并讨论了获得大量同步化发生的体细胞胚的原因。
Proloplasts wcre isolated from stemnode-derived embryogenic callus with an enzyme mixture containing 1.5% Cellu lase R-10 + 0.5% Macerozyme R-10 + 0.5% Snailasc + 0.5 mmol/L CaCl_2 + 0.6 mol/L mannitol + 0.3% potassium dextran sulfate at pH 5.8, and cultured in modified MS medium supplemented with 0.3% agarose and 1 mg/L 2,4-D + 0.5 mg/L zeatin. They divided with a high frequency, 40—50% at l0 days culture. A month later the protocolonics were transferred to MS liquid medium with 0.5 mg/ L 2,4-D and decreased osmoticum. After proliferated in large number, they were induced to differentiate by transferring to MS medium without hormone or containing 0.1 mg/L zeatin, where over 90% of them formed globular embryos synchronously and developed in succession into heart-shaped embryos with 70% of synchronization. Further development of the embryos resulted in the formation of whole plantlets.The sake for the success of getting a great number of somatic embryos and high synchronization in cmbryogenesis was also discussed in this paper.
出处
《实验生物学报》
CSCD
1991年第4期395-401,共7页
Acta Biologiae Experimentalis Sinica
关键词
峨参
原生质体
培养
体细胞
胚
Anthriscus sylvestris. Embryogencsis. Piant regeneration. Protoplast culture. Synchronization.
