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我们采用PEG—高pH—高钙的方法对大豆和豌豆原生质体进行融合,获得了10—36％的融合率。大豆原生质体游离的混合酶液为2％ Cellulase ONOZUKA R-10,2％ Hemicellulase-Rhozyme, 1％ Pectinase Sigma;豌豆原生质体游离的混合酶液为2％Cellulase ONOZUKA R-10,2％ Hemicellulase sigma,1％ Pectinase Sigma。原生质体的密度对融合的影响很大,以10~4—10~(?)/毫升的密度适合原生质体融合及有利细胞分裂。品种间融合率有差异。融合体用高国楠的KM8P培养基培养,经过多次分裂,获得了愈伤组织。
We fused the protoplasts of soybean and pea with PEG-high pH-high calcium, yielding 10-36% confluency. Soybean protoplast free mixed enzyme solution was 2% Cellulase ONOZUKA R-10,2% Hemicellulase-Rhozyme, 1% Pectinase Sigma; pea protoplast free mixed enzyme solution was 2% Cellulase ONOZUKA R-10,2% Hemicellulase sigma, 1% Pectinase Sigma. The density of protoplasts has a great influence on the fusion, and it is suitable for protoplast fusion and favorable cell division at the density of 10 ~ 4-10 ~ (?) / Ml. Differences in the rate of fusion between varieties. The fusion was cultured with KM8P medium of Gao Nan, and after several splits, the callus was obtained.