Abstract: Mesophyll protoplasts of wild Medicago falcata were isolated from young leaves (lower epi dermis was tored off)of 15~ 20 days old by mixed enzymatic solution containing cellulase 1%, hemicellu lase 1% and pectinase 0.5%.The protoplasts were cultured in dark,in thin-layer culture fluid of improverd MS medium with 6-BA lmg/L, 2, 4-D 0.05-0.5mg/L and NAA 1mg/L,and underwent the first time cell division after 4 day’ s culture.Through many times cell division,the regenerated cells formed about lmm (in diameter)multicellular clusters in changed culture fluid with lower osmotic pressure.The clusters de veloped into calli on agar solidified MS rnedium with KT and 2,4-D(the best ratio of KT and 2,4-D is 1:3),then the calli regenerated complete plant after 4-5 times subculture on differentiating medium.

Key words: Wild Medicago falcata, Protoplast, Culture, Regenerated plant

摘要： 取野生黄花苜蓿15～20天叶龄的叶片,去掉下表皮,用1%纤维素酶、1%半纤维素酶和0.5%果胶酶的混合液游离原生质体。用改良的MS培养基附加2,4－D、BA和NAA等,在黑暗条件下浅层悬浮培养。培养第4天、第7天和第10天,原生质体分别出现第一次、第二次和多次分裂,在此期间数次加液或换液调节渗透压,直至形成1mm左右的大细胞团,然后转移到附加2,4-D和KT等的MS固体培养基,诱导形成愈伤组织,再经4～5次继代培养,分化成为完整的再生植株。

关键词: 野生黄花苜蓿, 原生质体培养, 再生植株