X-Ray Film Gel Contact Print Method for Analyzing Differential Expression of Plant Protease Inhibitors Under Abiotic Stress

The regulation of proteolytic enzymes by protease inhibitors is crucial for maintaining the balance between protein synthesis and degradation, preventing uncontrolled proteolysis and fine-tuning cellular processes essential for optimal function and survival of the plants. It is known that the plant protease inhibitors activities are induced in defense of biotic as well as abiotic stresses. Thus, beyond their fundamental physiological functions, their involvement in stress responses, such as drought, cold, and salinity, is of equally significant. The X-ray film contact print method is an effective method for assessing various protease inhibitors exposed to stress conditions. In this approach, initially plant protease inhibitors will be separated using electrophoresis, and then the gel is treated with trypsin, which inhibits protease inhibitors. This gel when placed on X-ray film, the trypsin will digest the gelatin layer present on the film and the gelatinolytic activity stalled at the premises of protease inhibitors. This will provide the impression of the differentially expressed protease inhibitors in stress-treated plants.