Homoeostasis of glutathione (GSH) is crucial for plant survival and adaptability against stress. Despite the presence of complete Arabidopsis and rice genome sequence, the comprehensive analysis of the GSH metabolizing genes is still missing. This research concentrated on the comprehensive understanding of GSH metabolizing genes in two model plants—Arabidopsis and rice in terms of their subcellular localization, exon–intron distribution, protein domain structure, and transcript abundance. Expression profiling using the microarray data provided significant evidence of their participation in response to various abiotic stress conditions. Besides, some of these GSH metabolizing genes revealed their expression alteration in several developmental changes and tissue diversification. The presence of various stress-specific cis-regulatory elements in the promoter region of GSH metabolizing genes could be directly correlated with their stress-specific transcript alteration. Moreover, the application of exogenous GSH significantly downregulated GSH synthesizing genes and upregulated GSH metabolizing genes in Arabidopsis with few exceptions indicating a product-dependent regulation of GSH metabolizing genes. Interestingly, validation of rice GSH metabolizing genes in response to drought and salinity showed an almost similar pattern of expression in quantitative real-time as observed by microarray data. Altogether, GSH metabolizing members are a promising and underutilized genetic source for plant improvement that could be used to enhance stress tolerance in plants.