AimWe conduct a biogeographical assessment of orchids in a global biodiversity hotspot to explore their distribution and occurrences of local hotspots while identifying geographic attributes underpinning diversity patterns. We evaluate habitat characteristics associated with orchid diversity hotspots and make comparisons to other centres of orchid diversity to test for global trends. The ultimate goal was to identify an overall set of parameters that effectively characterize critical habitats to target in local and global orchid conservation efforts.LocationCosta Rica; Mesoamerica.TaxonOrchidaceae.MethodsData from an extensive set of herbarium records were used to map orchid distributions and to identify diversity hotspots. Hotspot data were combined with geographic attribute data, including environmental and geopolitical variables, and a random forest regression model was utilized to assess the importance of each variable for explaining the distribution of orchid hotspots. A likelihood model was created based on variable importance to identify locations where suitable habitats and unidentified orchid hotspots might occur.ResultsOrchids were widely distributed and hotspots occurred primarily in mountainous regions, but occasionally at lower elevations. Precipitation and vegetation cover were the most important predictive variables associated with orchid hotspots. Variable values underpinning Costa Rican orchid hotspots were similar to those reported at other sites worldwide. Models also identified suitable habitats for sustaining orchid diversity that occurred outside of known hotspots and protected areas.Main conclusionsSeveral orchid diversity hotspots and potentially suitable habitats occur outside of known distributions and/or protected areas. Recognition of these sites and their associated geographic attributes provides clear targets for optimizing orchid conservation efforts in Costa Rica, although certain caveats warrant consideration. Habitats linked with orchid hotspots in Costa Rica were similar to those documented elsewhere, suggesting the existence of a common biogeographical trend regarding critical habitats for orchid conservation in disparate tropical regions.

Pleurothallidinae (Epidendreae) are a megadiverse Neotropical orchid subtribe comprising > 5100 species, most of which are probably pollinated by Diptera. The role of pollinators as drivers of species diversity is largely unknown because knowledge of pollination systems in Pleurothallidinae is still scarce. Here, we addressed the pollination of Trichosalpinx s.s. through study of floral anatomy, pollinator behaviour and floral traits shared with other angiosperms to elucidate its pollination mechanisms. We identified midge specimens with DNA barcoding and morphology, documented pollination with video recordings, studied the anatomy of flowers by combining microscopy (light microscopy, scanning electron microscopy and transmission electron microscopy) and histochemistry and analysed floral scents with gas chromatography-mass spectrometry. We found that two Trichosalpinx spp. are pollinated exclusively by female biting midges of a Forcipomyia (Euprojoannisia) sp. (Ceratopogonidae). The midges land on the motile lip and appear to suck substances from its papillose surface. We detected secretion of carbohydrates and proteins on the lip and sepals, and thus, Trichosalpinx might stimulate a protein collection instinct in female biting midges. The well-developed mandibles and poorly developed laciniae of the pollinators indicate that they mainly feed on invertebrate hosts from which they draw haemolymph. Thus, Trichosalpinx flowers offer small quantities of proteins and carbohydrates that may act as flavour teases and together with the colour, fragrances, trichomes and movement of the lip, they probably form part of a complex deceptive system. Some other angiosperms that are also pollinated by biting midges possess similar dark purple flowers with ciliate ornamentation and use myophily, sapromyophily or kleptomyiophily as strategies to exploit different families of Diptera as pollinators. One Forcipomyia sp. (Euprojoannisia) is kleptoparasitic, suggesting that kleptomyiophily may have evolved in Trichosalpinx. The similar floral morphology among members of Trichosalpinx and some species of the closely related genera Anathallis and Lankesteriana suggests that they are also pollinated by biting midges.

We propose a new classification of the Lepanthes affinity based on previous studies and our ongoing phylogenetic re-evaluation of the Pleurothallidinae. Fourteen genera are recognized as belonging to the affinity. They are found highly supported in a DNA-based phylogenetic inference of combined plastid (matK) and nuclear (nrITS) datasets. The necessary changes, including four novel generic concepts, needed to reorganize the Lepanthes affinity, are proposed here to insure monophyly. The integral discussion on the phylogenetics and biogeography of the group, together with morphological characterization of each clade will be presented in a follow up study.