Alfalfa (Medicago sativa L.) grown in acidic soils is often affected by phosphorus (P) deficiency, which results in reduced mineral nutrient content and forage quality. In this context, the effects of phosphorus (P) fertiliser remain unclear. In this study, we analysed the effects of P application on mineral nutrient content and forage quality in aluminium (Al)-sensitive (Longzhong) and Al-tolerant (Trifecta) alfalfa cultivars cultivated in two acidic soil environments. Mineral nutrient content and quality were affected by genotype, soil type, and P treatment concentration (p < 0.001). In limestone soil, for Longzhong and Trifecta, the optimal potassium (K), calcium (Ca), and magnesium (Mg) contents as well as crude protein content (CP) and ether extract (EE) values were observed at 20 mg P kg−1, that of the P content was observed at 40 mg P kg−1, and the minimum neutral detergent fibre (NDF) acid detergent lignin (ADL) values were observed at 40 mg P kg−1. In yellow soil, the maximum K, Ca, Mg, and P contents in Longzhong and Trifecta were observed at 40 mg P kg−1, whereas the maximum CP, EE, and ADL values were observed at 20 mg P kg−1. Our study provides an empirically based framework for optimising alfalfa fertilisation programmes in acidic soils.