One of the main challenges faced by many rural areas is the presence of natural constraints, such as climatic conditions, steep slopes, and poor soil quality, which make agricultural production and activities more difficult and costly. In these areas, there is a significant risk of agricultural land abandonment, leading to potential losses in biodiversity, degradation of rural landscapes, desertification, and increased forest fire risk. The Common Agricultural Policy (CAP) aims to mitigate these risks through specific payment schemes provided to areas facing natural and other specific constraints. In this context, mapping and measuring territorial differentiation is essential for informing policy responses. At the end of the previous CAP programming period, the EU updated its classification of Less Favored Areas (LFAs), experimenting with a flexible approach based on common biophysical criteria (definitions and thresholds) and methodological guidelines to delineate territorial differentiations that are both locally relevant and comparable across member states. This contribution presents a review of the current state of data and spatial inference systems used in Italy to delineate biophysical limitations and assess the presence of factors that may help offset the impact of natural constraints. This process has supported the analysis of territorial differentiation and highlighted the related implications for agricultural entrepreneurs operating in diverse contexts.