<p>Soil tillage practices and fertilization are the backbones of potato crop production system. Sustainable agricultural practices are essential for enhancing potato production in the Mediterranean environment and addressing the food security challenges. The main goal of research is to evaluate the efficacy of different soil tillage practices in combination with different fertilization sources on two potato genotypes to enhance plant growth, yield, disease resistance, and soil characteristics in the Mediterranean environment over the period of two consecutive years. Three tillage practices were adopted: plowing as conventional tillage, subsoiling, and spading as conservation tillage. Two fertilization treatments included mineral and organic fertilization. The two potato varities cultivated were Levante and Agata. The total soil nitrogen (TN) percentage in the soil increased to 0.16% under subsoiling with organic fertilization, followed by improvements in total carbon percentage and carbon-to-nitrogen (C/N) ratio. However, these practices did not significantly impact potato yield. The highest yield of commercial tubers was observed with plowing (27.3 n. m^− 2) and mineral fertilization (25.3 n. m^− 2). For seed tubers, spading tillage (34.1 n. m^− 2) and organic fertilization (33.8 n. m^− 2) were effective in improving both yield and quality. The Levante potato variety, The 4th International Electronic Conference on Agronomy showed resistance to early blight (<i>Alternaria</i> spp.) and root rot (<i>Sclerotium rolfsii</i>), in contrast to the Agata variety. Implementing conservation tillage practices combined with organic fertilization and disease resistant potato varieties may enhance sustainability in potato production in Mediterranean environments.</p>