This study investigated the potential of gamma radiation to enhance the quality of dwarf napier grass (Pennisetum purpureum cv. Mot) cultivated in acidic soil, with a focus on its macro-mineral content and rumen fluid characteristics. The research findings focus on the impact of gamma radiation solely on macro minerals and rumen fluids, enhancing our insights into innovative approaches for improving livestock nutrition in regions with prevalent acidic soils. The experiment involved the application of various gamma radiation doses (0, 5, 10, 15, 20, 25, and 30 Gy). Four replications were done using a randomized block design, with 25 Pennisetum purpureum cv. Mot seeds in each repetition, totalling 700 seeds. The plants were grown in acidic soil with a pH of approximately 4.5-5. After two months of growth, the plants were harvested, and various parameters were analyzed. The results of the research indicated that the application of a 15 Gy dosage significantly enhanced the absorption of P (p<0.01), N (p<0.01), and Ca (p<0.01) in the plant. Additionally, gamma irradiation at a 25 Gy dosage demonstrated a notable effect, resulting in the highest calcium (Ca) content (p<0.01). However, the treatment did not exhibit a significant impact on pH (p > 0.05) in the rumen fluid, while NH3 (p<0.01) and VFA (p<0.01) parameters in the rumen fluid were significantly affected. The optimal dosage for the rumen fluid parameters NH3 and VFA was found to be 15 Gy. Overall, it can be concluded from this study that gamma irradiation doses of 15 Gy can enhance the nutritional content of Pennisetum purpureum cv. Mot cultivated on acidic soil, particularly when evaluated based on mineral content and rumen fluid parameters.