Floral odour chemistry defines species boundaries and underpins strong reproductive isolation in sexually deceptive orchids.
OBJECTIVE
The events leading to speciation are best investigated in systems where speciation is ongoing or incomplete, such as incipient species. By examining reproductive barriers among incipient sister taxa and their congeners we can gain valuable insights into the relative timing and importance of the various barriers involved in the speciation process. The aim of this study was to identify the reproductive barriers among sexually deceptive orchid taxa in the genus Chiloglottis.


METHODS
The study targeted four closely related taxa with varying degrees of geographic overlap. Chemical, morphological and genetic evidence was combined to explore the basis of reproductive isolation. Of primary interest was the degree of genetic differentiation among taxa at both nuclear and chloroplast DNA markers. To objectively test whether or not species boundaries are defined by the chemistry that controls pollinator specificity, genetic analysis was restricted to samples of known odour chemistry.


RESULTS
Floral odour chemical analysis was performed for 600+ flowers. The three sympatric taxa were defined by their specific chiloglottones, the semiochemicals responsible for pollinator attraction, and were found to be fully cross-compatible. Multivariate morphometric analysis could not reliably distinguish among the four taxa. Although varying from very low to moderate, significant levels of genetic differentiation were detected among all pairwise combinations of taxa at both nuclear and chloroplast loci. However, the levels of genetic differentiation were lower than expected for mature species. Critically, a lack of chloroplast DNA haplotype sharing among the morphologically indistinguishable and most closely related taxon pair confirmed that chemistry alone can define taxon boundaries.


CONCLUSIONS
The results confirmed that pollinator isolation, mediated by specific pollinator attraction, underpins strong reproductive isolation in these taxa. A combination of large effective population sizes, initial neutral mutations in the genes controlling floral scent, and a pool of available pollinators likely drives diversity in this system.
Evolution, Ecology and Genetics, Research School of Biology, The Australian National University, Canberra, ACT 0200, Australia.
Rod
R
Peakall
Michael R
MR
Whitehead
D001315:Australia; D018742:DNA, Chloroplast / Q000235:genetics; D035264:Flowers / Q000033:anatomy & histology / Q000737:chemistry* / Q000235:genetics; D056426:Genetic Loci; D006239:Haplotypes / Q000235:genetics; D018895:Microsatellite Repeats / Q000235:genetics; D015999:Multivariate Analysis; D009812:Odorants / Q000032:analysis*; D029595:Orchidaceae / Q000033:anatomy & histology / Q000235:genetics / Q000502:physiology*; D064209:Phytochemicals / Q000737:chemistry; D060047:Reproductive Isolation; D013045:Species Specificity
D018742:DNA, Chloroplast; D064209:Phytochemicals
2,5-dialkylcyclohexane-1,3-diones; Chiloglottis; Orchidaceae; chloroplast DNA; cpSSRs; floral odour; microsatellites; orchid; pollination; pollinator-driven speciation; population genetic analysis; sexual deception