Agroecosystems show spatio-temporal variation that generates “pulses” in biomass production throughout the landscape, likely affecting the use of space and the trophic relationships of wild species. Plant biomass is considered a primary source of energy; thus, it can influence species composition and abundance. This study aimed to evaluate the relationship between above-ground biomass and patterns of diversity, distribution, and abundance of non-volant small mammals in the main agro-silvicultural landscapes of São Paulo state in southeast Brazil: sugarcane and Eucalyptus plantations, with fragments of native vegetation. From October 2021 to September 2022, we carried out monthly sampling campaigns over a grid of 300 traps in a Eucalyptus-dominated landscape and 240 traps in a sugarcane-dominated landscape, adapted from RAPELD methodology. Remote images taken by unmanned aerial vehicles (UAV) of native vegetation and abandoned pasture were used to estimate above-ground biomass. Within agrosilvicultural landscapes, an increase in biomass prompts a shift in the abundance of small mammals, transitioning from open area species to forest species. Particularly, Akodon montensis, Calomys tener, Didelphis albiventris, Nectomys squamipes and Oligoryzomys nigripes show potential as ecological indicators of processes within agro-silvicultural landscapes. This study supports the use of native vegetation biomass as a proxy for assessing small mammals’ abundance in agrosilvicultural landscapes. Moreover, native vegetation recovery emerges as one of the most effective strategies for enhancing the conservation value of agrosilvicultural landscapes.