Habitats and ecological communities occurring in the mesophotic region of the central Great Barrier Reef (GBR), Australia, were investigated using autonomous underwater vehicle (AUV) from 51 to 145 m. High-resolution multibeam bathymetry of the outer-shelf at Hydrographers Passage in the central GBR revealed submerged linear reefs with tops at 50, 55, 80, 90, 100 and 130 m separated by flat, sandy inter-reefal areas punctuated by limestone pinnacles. Cluster analysis of AUV images yielded five distinct site groups based on their benthic macrofauna, with rugosity and the presence of limestone reef identified as the most significant abiotic factors explaining the distribution of macrofaunal communities. Reef-associated macrofaunal communities occurred in three distinct depth zones: (1) a shallow (<60 m) community dominated by photosynthetic taxa, notably scleractinian corals, zooxanthellate octocorals and photosynthetic sponges; (2) a transitional community (60–75 m) comprising both zooxanthellate taxa and azooxanthellate taxa (notably gorgonians and antipatharians); and (3) an entirely azooxanthellate community (>75 m). The effects of depth and microhabitat topography on irradiance most likely play a critical role in controlling vertical zonation on reef substrates. The lower depth limits of zooxanthellate corals are significantly shallower than that observed in many other mesophotic coral ecosystems. This may be a result of resuspension of sediments from the sand sheets by strong currents and/or a consequence of cold water upwelling.