Understanding environmental and spatial gradient influences on sediment microbial communities, especially the communities of highly contaminated subsurface sediments, has received great attention with respect to natural attenuation and bioremediation. Here, we investigated the spatial variation and the co-occurrence patterns of microbial communities in polycyclic aromatic hydrocarbon (PAH)-contaminated riverine sediments by using spatial-series 16S rRNA gene data. The results showed that species from the surface and subsurface sediment samples tended to show greater co-occurrence patterns and facilitative interactions in the sediment microbial community as environmental severity increased. Microorganisms in the heavier PAH-contaminated sediment have stronger relationships and are more centrally clustered within the network compared to microorganisms in the lower PAH-contaminated sediment. The core communities harbored the keystone species (Dechloromonas, Crenothrix, Desulfitromonadales, Xanthomonadales, Anaerolineaceae and Dehalococcoidales), which responded to changes in the environmental and spatial gradients. The sediment PAH concentrations, ferrous iron and vertical distance were identified as the main drivers in determining the bacterial community assembly. The keystone species were linked to PAHs biodegradation coupled with iron cycling in sediments and could orchestrate core communities to perform ecosystem processes. Overall, these findings provide new insight into microbial community assembly and contribute to harnessing their functions in ecosystems for bioremediation.