Nowadays we are seeing the largest biodiversity loss since the extinction of the dinosaurs. To conserve biodiversity it is essential to plan protected areas using a prioritization approach, which takes into account the current biodiversity value of the sites. Considering that in the Mediterranean Basin the agro-ecosystems are one of the most important parts of the landscape, the conservation of crops is essential to biodiversity conservation. In the framework of agro-ecosystem conservation, farmland birds play an important role because of their representativeness, and because of their steady decline in the last Century in Western Europe. The main aim of this research was to define if crop dominated landscapes could be useful for biodiversity conservation in a Mediterranean area in which the landscape was modified by humans in the last thousand years and was affected by the important biogeographical phenomenon of peninsula effect. To assess this, we identify the hotspots and the coldspots of bird diversity in southern Italy both during the winter and in the breeding season. In particular we used a scoring method, defining a biodiversity value for each cell of a 1-km grid superimposed on the study area, using data collected by fieldwork following a stratified random sampling design. This value was analysed by a multiple linear regression analysis and was predicted in the whole study area. Then we defined the hotspots and the coldspots of the study area as 15% of the cells with higher and lower value of biodiversity, respectively. Finally, we used GAP analysis to compare hotspot distribution with the current network of protected areas. This study showed that the winter hotspots of bird diversity were associated with marshes and water bodies, shrublands, and irrigated crops, whilst the breeding hotspots were associated with more natural areas (e.g. transitional wood/shrubs), such as open areas (natural grasslands, pastures and not irrigated crops). Moreover, the results underlined the negative effects of permanent crops, such as vineyards, olive groves, and orchards, in particular during the winter season. This research highlights the importance of farmland areas mainly for wintering species and the importance of open areas for breeding species in the Mediterranean Basin. This may be true even when the species’ spatial distribution could be affected by biogeography. An important result showed that the hotspots for breeding species cannot be used as a surrogate for the wintering species, which were often not considered in the planning of protected areas.