This study was conducted to investigate the influence of gamma rays on stomatal parameters and the interaction of these traits with agronomy of the sugarcane crop. Three genotypes of sugarcane (NIA-0819, NIA-98, and BL4) were exposed to four doses of gamma radiation (10, 20, 30, and 40 Gy) and then subjected to field trials. Stomatal length, width, and density were observed using scanning electron microscopy. Moreover, agronomic and sugar-related characteristics of the crop were determined at maturity. The stomatal parameters augmented at two lower doses of gamma radiation (10 and 20 Gy) and declined under higher doses (30 and 40 Gy). The maximum stomatal length was observed in NIA-0819 at 10 Gy (63.71 µm), whereas NIA-98 and BL4 demonstrated highest stomatal length under 20 Gy (54.11 and 57.40 µm, respectively), indicating a role of genetic factors in varietal response. Similar trend was noticed for stomatal width and density as well. The lowermost treatment (10 Gy) of NIA-0819 produced maximum stomatal density (115.31 stomata mm-2 on the abaxial surface). Adaxial stomatal density was significantly lower than the abaxial one. Sugar quality attributes revealed a different tendency. Sucrose contents of BL4 increased gradually from 12.33% at control to 14.54% at 40 Gy. Yield and yield-contributing traits of genotypes indicated a strong positive correlation with the stomatal parameters. The present study suggested that gamma radiations cause variations in stomatal characteristics of sugarcane. These changes further influence the photosynthetic activity and instigate a direct impact on the agronomic performance of the crop.