Microplate quantification of enzymes of the plant ascorbate-glutathione cycle.
Here, we describe microplate assays for determining the specific activities of four enzymes that constitute the ascorbate-glutathione cycle: APX, MDHAR, DHAR, and GR. In plants, these enzymes play a major role in detoxifying reactive oxygen species produced in cells under environmental stress. This work presents the development of plate reader assays to allow rapid analysis of the ascorbate-glutathione cycle activity using tomato fruits subjected to salt stress as a model. With this method, it is possible to analyze easily in one day the activities of the four enzymes for 30 experimental samples, all in triplicate and with blanks.
Murshed
Ramzi
R
Université d'Avignon et des Pays de Vaucluse, Laboratoire Physiologie des Fruits et Légumes, F-84018 Avignon, France.
Lopez-Lauri
Félicie
F
Sallanon
Huguette
H
D060387:Ascorbate Peroxidases; D001205:Ascorbic Acid / Q000378:metabolism*; D005638:Fruit / Q000201:enzymology; D005978:Glutathione / Q000378:metabolism*; D005980:Glutathione Reductase / Q000378:metabolism; D018551:Solanum lycopersicum / Q000201:enzymology*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D010088:Oxidoreductases / Q000378:metabolism; D010544:Peroxidases / Q000378:metabolism
D010088:Oxidoreductases; D010544:Peroxidases; D060387:Ascorbate Peroxidases; D009247:NADH, NADPH Oxidoreductases; C023927:monodehydroascorbate reductase (NADH); D005980:Glutathione Reductase; C020666:glutathione dehydrogenase (ascorbate); D005978:Glutathione; D001205:Ascorbic Acid