Pollinator-mediated isolation promotes coexistence of closely related food-deceptive orchids.

Identifying the factors that contribute to reproductive isolation among closely-related species is key for understanding the diversification of lineages. In this study, we investigate the strength of premating and postmating reproductive isolation barriers between Disa ferruginea and Disa gladioliflora, a pair of closely-related species, often found co-flowering in sympatry. Both species are non-rewarding and rely on mimicry of different rewarding model flowers for attraction of pollinators. We constructed abiotic niche models for different forms of each taxon to measure ecogeographic isolation. Using experimental arrays in sympatry, we recorded pollinator transitions to measure ethological isolation. We performed hand pollinations to measure postpollination isolation. We found strong, but not complete, premating isolation associated with abiotic niches and absolute ethological isolation based on pollinator preferences in sympatry. Pollinator preferences among the orchids could be explained largely by flower colour (orange in D. ferruginea and pink in D. gladioliflora) which matches that of the pollinator food plants. Post-mating barriers were weak as the species were found to be inter-fertile. Coexistence in the orchid species pair is due mainly to ethological reproductive isolation arising from flower colour differences resulting from mimicry of different rewarding plants. These results highlight the importance of signalling traits for ethological isolation of closely-related species with specialized pollination systems.