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试验56份大豆栽培品种。采用幼荚子叶、无菌苗下胚轴和子叶、幼胚悬浮培养细胞作原生质体游离材料。苗期子叶和下胚轴用酶液:1％Hemicellulase HP150,0.4％Cellulase R—10,0.1％Pectolyase Y—23;幼荚子叶用1％Cellulasc R—10,1％Hemicellulase HP150,0.5％Pectinase;悬浮培养细胞用1％Cellulase R—10,0.2％Pectolyase Y—23,2％Driselase游离,4种材料均获得大量原生质体。原生质体培养基均为K8P,分化培养基为MS、B5,培养基中的激素试用了2.4—D,2.4.5—T,NAAIAA、ZT、KT、BA、zip和毒莠定等不同配合和浓度。结果有12份品种的幼荚子叶。下胚轴和苗期子叶的原生质体形成愈伤组织、有不同类型的根分化和绿点出现,分化培养基B5+0.1-0.2mg/l 2.4.5-T+1-1.5mg/l·BA较好。
56 soybean cultivars were tested. Using the young pod cotyledons, hypocotyls and cotyledons of sterile seedlings, the suspension cultures of immature embryos were used as protoplast free material. Seedling leaf and hypocotyl with enzyme solution: 1% Hemicellulase HP150, 0.4% Cellulase R-10, 0.1% Pectolyase Y-23; young pod leaves with 1% Cellulasc R- 10, 1% Hemicellulase HP150, 0.5% Pectinase; Suspension cultured cells were dissociated with 1% Cellulase R-10, 0.2% Pectolyase Y-23 and 2% Driselase. A large number of protoplasts were obtained from all four kinds of materials. Protoplast culture medium was K8P, and differentiation medium was MS, B5. 2.4-D, 2.4.5-T, NAAIAA, ZT, KT, BA, zip and picloram were used in different combinations and concentrations . Results are 12 varieties of young Podocarpus. Hypocotyls and seedlings cotyledons protoplasts formed callus, there are different types of root differentiation and green spots appear, differentiation medium B5 + 0.1-0.2mg / l 2.4.5-T + 1-1.5mg / l · BA is better.