【摘要】 外植体类型和光照条件决定沙打旺胚性愈伤组织的形成。用生长10d的胚性愈伤组织可分离到12×106个/g(原生质体/细胞),活力超过80%。当原生质体以10×105/mL的植板密度培养在含06%琼脂糖附加15mg/L2,4D、05mg/LBA和05mol/L葡萄糖的培养基(无机盐降为1/4)中,植板率为168%。条件培养基显著促进原生质体的生长发育。长大的细胞克隆经2周4℃低温处理后转到含01mg/LNAA和10mg/LBA分化培养基上,体细胞胚胎发生频率高达70%,每克细胞产生的体细胞胚数在200个以上。成熟的体细胞胚转到无激素的1/2MS培养基中即分化成苗,再生植株为正常的二倍体。更多还原

【Abstract】 Embryogenic callus was obtained only from hypocotyl explants of Astragalus adsurgens and light inhibited the formation of embryogenic callus.A high yield (1 2×10 6/g F.Wt.) of protoplasts with high viability (over 80%) could be isolated from 10 day old embryogenic callus.Protoplasts were induced to undergo sustained division and to form cell colonies when cultured in agarose solidified medium (KMP) containing 1/4 strength of mineral salts and supplemented with 1 5mg/L 2,4 D,0 5mg/L BA and 0 5mol/L glucose at a plating density of 1 0×10 5mL,where the plating efficinency was 16 8%.Conditioning medium significantly improved the formation of cell colonies.When protoplast derived colonies were maintained at 4℃ for 2 weeks and subsequently transferred onto medium (MS) with 0 1mg/L NAA and 1 0mg/L BA,somatic embryogenesis occurred.Frequency of cell colonies producing somatic embryos reached 70%,and the number of somatic embryos per gram cells was over 200.Cultured on hormone free half strength MS medium,somatic embryos developed into healthy plantlets with normal chromosome complement.更多还原