Nishimura M, Douce R, Akazawa T
Research Institute for Biochemical Regulation, School of Agriculture, Nagoya University, Chikusa, Nagoya 464, Japan.
Plant Physiol. 1985 Jun;78(2):343-6. doi: 10.1104/pp.78.2.343.
A method was developed for the quantitative analysis of intactness of spinach leaf protoplasts using glycolate oxidase activity as an index. Since glycolate does not penetrate into protoplasts at neutral pH, the increase of O(2) consumption by the addition of glycolate to protoplast suspension was due to the glycolate oxidase activity released from damaged protoplasts. The proportion of damaged protoplasts in the whole preparation was calculated from the ratio of released and total glycolate oxidase activity. Freshly prepared spinach leaf protoplasts were found to be 80 to 90% intact as estimated by the method. The effect of osmolarity on the respiratory activities of spinach leaf protoplasts was also examined by applying the same principle.
开发了一种以乙醇酸氧化酶活性为指标定量分析菠菜叶原生质体完整性的方法。由于在中性pH值下乙醇酸不能穿透原生质体，向原生质体悬浮液中添加乙醇酸后氧气消耗的增加是由于受损原生质体释放的乙醇酸氧化酶活性所致。根据释放的和总的乙醇酸氧化酶活性的比例计算整个制剂中受损原生质体的比例。通过该方法估计，新鲜制备的菠菜叶原生质体完整率为80%至90%。应用相同原理还检测了渗透压对菠菜叶原生质体呼吸活性的影响。
