刺孢吸水链霉菌与淡紫灰链霉菌原生质体制备条件研究

Conditions about protoplast preparation of S.hygrospinous var.Beijingensis and S.lavendule

通过对甘氨酸浓度、酶解温度、酶浓度、酶解时间等影响因素考察,及正交试验优化,确定了刺孢吸水链霉菌和淡紫灰链霉菌原生质体制备的最佳条件.实验结果表明,刺孢吸水链霉菌最佳原生质体制备条件是:甘氨酸浓度为0.5%,酶解温度28℃,溶菌酶浓度为3×10-3g/mL,酶作用时间60 min;淡紫灰链霉菌:甘氨酸浓度为1.0%,酶解温度32℃,溶菌酶浓度4×10-3g/mL,酶作用时间60 min.有效的原生质体保存条件是:-20℃,在5 d内,原生质体再生率在15%以上.

Conditions about protoplast optimum preparation of S. hygrospinous var. Beijingensis and S. lavendule were studied by orthogonal test, including the conditions of concentration of Gly, lytic temperature, lysozyme concentration and lytic time. Experimental results indicated that the conditions of optimum preparation of S. hygrospinous var. Beijingensis are as following： Gly 0.5 %, lytic temperature 28 ℃, lysozyme 3 × 10^-3 g/mL,lytic time 60 min. Those of S. lavendule are as following： Gly 1.0%, lytic temperature 32 ℃, lysozyme 4 × 10^-3 g/mL, lytic time 60 min. The regenerated frequency of protoplasts could reach up to above 15% in the conditions of -20 ℃ ,5 days.