Biodiversity is essential for the functioning of ecosystems and the provision of services. In recent years, the role of plantations in mitigating climate change through carbon sequestration has been highlighted. In the Mediterranean area, high-density poplar plantations in short-rotation with resprouting management (SRC) have been established for biomass purposes on mostly irrigated agricultural land, coexisting with rainfed and irrigated agricultural crops. This study aims to assess the contribution of these plantations to this type of agroforest ecosystem in terms of biodiversity. For this purpose, both flora and fauna diversity were evaluated both within and outside of the plantation. Additionally, the accumulated carbon in the biomass, as well as in the accompanying vegetation within the plantation, was assessed. Different indices were used to evaluate both the intrinsic diversity of the forest plantation and the degree of substitution and complementarity between the different communities of the landscape. Our findings reveal distinct biodiversity patterns in the land-use scenarios sampled. Specifically, we observed significantly higher flora-species richness in SRC plantations than in the adjacent agricultural land, whereas fauna richness showed a similar but slightly higher level in the forested area. A moderate level of complementarity between land uses was found for insects and mammals (around 45 %), contrasting with high complementarity for birds (87 %) and flora (90 %). This suggests substantial turnover and replacement among these ecological environments. Our results indicate that a second rotation (4 year) plantation could accumulate a total of 61.6 Mg C ha−1, and even though adventitious flora represents <2 % of the total carbon accumulated, its importance in providing ecosystem services is considerable. Hence, these findings evidence the fact that SRC poplar plantations can enhance biodiversity in Mediterranean agroforest ecosystems and actively contribute to various provisioning ecosystem services, including carbon sequestration, reflecting a multi-objective approach that extends beyond biomass production.