紫锥菊愈伤组织原生质体分离方法初探

Protoplast Isolation of Callus in Echinacea augustifolia

紫锥菊提取物属于目前国际上流行的草本植物药,其主要运用于增强免疫力等方面.以加拿大狭叶紫锥菊愈伤组织为材料,研究影响其原生质体分离的因素,结果表明选用浅黄绿色、质地均匀且呈疏松颗粒状的紫锥菊愈伤组织易游离出原生质体;采用纤维素酶浓度为2.0%,果胶酶浓度为1.0%,半纤维素酶浓度为0.5%,甘露醇浓度为0.7 mol.L-1的酶液配方酶解时间为8 h时,原生质体的产量为每克鲜重含50.0×104个.由此归纳出一套完整的愈伤组织诱导,组织培养,原生质体分离纯化,原生质体鉴定及计数的方法.

Preparations from Echinacea are among the most widely used herbal medicines. Most uses of Echinacea are based on the reported immunological properties. The authors use callus of Echinacea augustifolia for isolation and researched the factors influencing the process of protoplasts preparation. The result indicates it is easy to isolate protoplast from buff-green Callus in Echinacea augustifolia which is looked like granule with Symmetrical character. The result also shows that the best enzyme solution concentration is composed of CeUulase2.0% （ w/v）, Pectinasel. 0% （ w/v）, HemiceUulase 0.5% （w/v）, 0.7 mol L^-1 mannitol, and 50.0 × 104/gFW protoplasts are obtained after 8 hours later. Finally, they summarized an integrated approach including caUus induction, callus culture, isolation, purification, identification and calculation of protoplasm.