Ficus carica L. is a fruit crop of notable nutritional and economic value. The ‘Xinjiang Zaohuang’ cultivar, rich in flavonoids, also holds considerable medicinal potential. To address the constraints of conventional propagation for mass production, this study developed an efficient and genetically stable generation protocol using healthy sprouted branches. MS medium was identified as the most effective basal medium for shoot growth. The highest adventitious bud induction rate (89.67%) and the greatest mean bud number (6.29) were achieved when explants were cultured on MS medium supplemented with 1 mg/L 6-BA and 0.1 mg/L IBA. In the organogenesis process, indole-3-butyric acid (IBA) promoted direct shoot formation with minimal callus intervention compared to naphthaleneacetic acid (NAA). The optimal combination for shoot elongation was 0.1 mg/L 6-BA and 0.01 mg/L IBA, which produced morphologically uniform shoots. For rooting, an IBA concentration of 1 mg/L was optimal, achieving a 96.7% success rate. Inter-simple sequence repeat (ISSR) analysis confirmed the genetic stability of all regenerated plants. These findings establish a reliable technical framework for the large-scale propagation of this valuable fig cultivar.