Extraction, Storage Duration, and Storage Temperature Affect the Activity of Ascorbate Peroxidase, Glutathione Reductase, and Superoxide Dismutase in Rice Tissue.

Abstract

In experimental plant science, research often faces large numbers of tissue samples resulting from sequential harvests of a larger number of genotypes and treatments combinations. Analyses of biological processes such as enzyme kinetics are often time-consuming or need specific sample preparation procedures before the actual measurements can be done. Time is thus often the critical factor and the possibility to store plant samples either as tissue or as extracts increases the available timeframe for analyses. Biological molecules such as enzymes often change their activities when stored and thus do not reflect the processes occurring in living tissue. We investigated the effect of different storage methods such as freeze-drying, freezing at −20 °C, and freezing at −80 °C on the activity of three enzymes known as antioxidants, namely ascorbate peroxidase, glutathione reductase, and superoxide dismutase from two rice varieties. Varieties differed in enzyme activity in extracts of fresh material from leaf blades, leaf sheaths, and roots. When subjected to different storage methods, there were no differences between varieties, but strong effects of the different storage methods on enzyme activities were found. The effects of the storage methods on enzyme activity strongly differed between extracts from stored tissue samples or extracts stored from freshly sampled material. We propose enzyme-specific storage methods and durations that allow for expanding the window for analyses in large experimental studies involving destructive samplings for enzyme kinetics.