Noctor G, Foyer C H
Department of Environmental Biology, Institute of Grassland and Environmental Research, Plas Gogerddan, Ceredigion, SY23 3EB, United Kingdom.
Anal Biochem. 1998 Nov 1;264(1):98-110. doi: 10.1006/abio.1998.2794.
The recent production of transformed plants with enhanced capacity for glutathione synthesis has highlighted the interactions between foliar glutathione and turnover of free amino acid pools. The development of a convenient method for simultaneous measurement of glutathione, gamma-glutamylcysteine, and 16 amino acids is reported. This method utilizes derivatization of compounds with o-phthalaldehyde in the presence of 2-mercaptoethanol followed by separation using reversed-phase high-performance liquid chromatography. Eluted compounds are detected fluorimetrically. The method was tested using untransformed poplars and poplars in which foliar thiol contents have been enhanced by overexpression of gamma-glutamylcysteine synthetase. Foliar contents of glutathione determined by this method were comparable to those measured in common extracts by two other techniques.
最近培育出的谷胱甘肽合成能力增强的转基因植物，凸显了叶片谷胱甘肽与游离氨基酸库周转之间的相互作用。本文报道了一种同时测定谷胱甘肽、γ-谷氨酰半胱氨酸和16种氨基酸的简便方法。该方法利用邻苯二甲醛在2-巯基乙醇存在下对化合物进行衍生化，然后采用反相高效液相色谱进行分离。洗脱后的化合物通过荧光法进行检测。该方法用未转化的杨树以及通过γ-谷氨酰半胱氨酸合成酶过表达使叶片硫醇含量增加的杨树进行了测试。用该方法测定的叶片谷胱甘肽含量与通过其他两种技术在普通提取物中测得的含量相当。
