玉米叶片原生质体的制备

Preparation of Protoplasts from Etiolated Maize Leaves

本文以玉米黄化叶片为材料，采用两步酶解的方法研究了从玉米叶片制备原生质体的方法，发现第一次酶解以３０℃处理１ｈ为宜，第二次酶解以３０℃处理２．５～３ｈ较合适。同时发现所得原生质体净化悬浮时用２５％的蔗糖溶液在２５００ｒｐｍ离心５ｍｉｎ能获得较好的净化效果。

By the methed of two steps of enzymes hydrolysis, the protoplasts were prepared from etiolated maize leaves. It was found that for the first step of enzymes hydnolysis,it was suitable that leaves were incubated with cellulase(0. 7%) and pectinase (0. 5% ) for 1 hour at 30℃, for the second step of enzymes hydrolysis, the suitable incubation duration was 2. 5 to 3 hours at 30℃ in above enzymes solution, It was also found that the purified protoplasts could be obtaind officiently from the supernatant through suspended by 25% solution of sucrose and centrifuged at 2 500 rpm for 5 minutes.