In the wine sector, the importance of producing sustainably and obtaining a genuine product has been well known for years, just as organic and agroecological farming practices are widespread and have become a priority for most farms. On the other hand, sustainability assessments, are often carried by using tools such as life cycle assessments, carbon, and water footprints, which do not consider certain ecological and socio-economic aspect and cannot properly communicate sustainability performance to consumers and society at large. In this paper the framework AgroEnvironmental Sustainability Information System (AESIS) was applied, utilizing a set of indicators suitable to evaluate different dimensions of an agroecosystem (physical, ecological, productive, and social) and providing data useful for the sustainability assessment of two case studies located in Tuscany and Lombardy regions, Italy. In the case study located in Tuscany represented by Tenuta Casadei, organic and biodynamic-based farming method named BioIntegrale (BIOINT) was evaluated, while in the case study located in Lombardy represented by Castello di Cigognola, conventional farming (CONV) was compared with organic combined with precision farming (ORGP). The assessment of those farming systems was carried out using a set of agro-environmental indicators based on a conceptual framework for the evaluation of sustainability of land use options. CONV, ORGP and BIOINT were compared with different management systems or techniques and with scientifically determined sustainability thresholds, targets, or politically determined critical limits identified for each single indicator. Although the comparison layout was not the same for both case studies, results highlighted an improvement gradient in terms of sustainability performance in the order CONV < ORGP < BIOINT. BIOINT showed a 92% of compliance with thresholds, ORGP showed a lower but still virtuous level of compliance with thresholds, i.e. 87%, while CONV was considerably worse with 47% of indicators performing above thresholds. Nevertheless, several agroecological based interventions might be carried out within both case studies to enhance sustainability within ecological and social dimension at Castello di Cigonola and within physical dimension at Tenuta Casadei, respectively. As it concerns the method, AESIS provided useful information on the state of sustainability which give several entry-point for actions to farmers.