Metacommunity theory has advanced scientific understanding of how species interactions and spatial processes influence patterns of biodiversity and community structure across landscapes. While the central tenets of metacommunity theory have been promoted as pivotal considerations for conservation management, few field experiments have tested the validity of metacommunity predictions. Here, we tested one key prediction of metacommunity theory—that decreasing habitat connectivity should erode metacommunity structure by hindering species movement between patches. For 2 years, we manipulated an experimental old‐field grassland ecosystem via mowing to represent four levels of habitat connectivity: (1) open control, (2) full connectivity, (3) partial connectivity, and (4) no connectivity. Within each treatment plot (10 × 10 m, n = 4 replicates), we measured the abundance and diversity (i.e., alpha and beta) of both flying and ground arthropods using sticky and pitfall traps, respectively. We found that the abundance and diversity of highly mobile flying arthropods were unaffected by habitat connectivity, whereas less mobile ground arthropods were highly impacted. The mean total abundance of ground arthropods was 2.5× and 2× higher in the control and partially connected plots compared to isolated patches, respectively. We also reveal that habitat connectivity affected the trophic interactions of ground arthropods, with predators (e.g., wolf spiders, ground spiders) being highly positively correlated with micro‐detritivores (springtails, mites) but not macro‐detritivores (millipedes, isopods) as habitat connectivity increased. Together these findings indicate that changes in habitat connectivity can alter the metacommunity structure for less mobile organisms such as ground arthropods. Because of their essential roles in terrestrial ecosystem functioning and services, we recommend that conservationists, restoration practitioners, and land managers include principles of habitat connectivity for ground arthropods when designing biodiversity management programs.