摘要
栽培大豆(Glycine max L.)的6个品种(“中豆5号”、“合丰22”、“黑龙26”、“保黑选三”、“清远青”和“吉林16”)和野生大豆(G.soja)的1个品系(SH-1)的未成熟子叶分离原生质体,培养在含0.2mg/l 2,4-D,1mg/l NAA 和0.5mg/l ZT 的 K_8P 液体培养基中,原生质体培养3—5天后,开始第一次分裂,其后持续分裂,在6周内形成大量的多细胞团和小愈伤组织。小愈伤组织在用 gelrite 固化的 K_3培养基上进一步生长到2—3mm 时,转到含0.05—0.1mg/l 2,4-D 或0.2mg/l NAA,0.5mg/l BA 的 MSB 培养基上,得到结构紧密的瘤状愈伤组织。这种愈伤组织转到含0.15mg/lNAA,BA、KT 和 ZT 各为0.5mg/l 的 MSB分化培养基上,可诱导形成芽,进而再生植株。近二年来,相继从栽培大豆的上述6个栽培品种和野生大豆的1个品系的原生质体分别获得了再生植株,移栽于土中后均能正常开花结实,现已获得子二代正常成熟种子。
Protoplasts were isolated from immature cotyledons of six cultivars of Glycine max L.and SH-1 strain of Glycine soja and initially cultured in the K_(8p) liquid medium supplemented with 0.2 mg/l 2,4-D,1 mg/l NAA and 0.5 mg/l ZT.The protoplasts started to divide after 3-5 days of culture.Sustained divisions resulted in mass production of cell colonies and small calli in 6 weeks.The calli further grew to 2-3 mm on the gelrite-solidified K_8 medium and were transferred onto the MSB medium with 0.05-0.1mg/1 2,4-D and 0.5 mg/l BA,to obtain compact and nodular calli.Shoot formation was initiated on MSB medium containing 0.15 mg/l NAA,05 mg/l each of BA,KT and ZT with or without 500 mg/1 CH.Whole plants were regenerated upon transferring 3-4 cm shoots to 1/2MS medium with 0.2 mg/l IBA.So far,128 complete plants have been obtained from 6 cultivars of Glycine max L.and 24 plants have been regenerated from SH-1 strain of Glycine soja,and normal seeds were obtained from them after transplanting into pots.
关键词
大豆
原生质体培养
植株再生
Cultivated soybean(Glycine max L.)
Wild soybean(Glycine soja)
Protoplast culture
Plant regeneration
