Polyploidy has played a key role in plant evolution and diversification. Despite this, the processes governing reproductive isolation among cytotypes growing in mixedploidy populations are still largely unknown. Theoretically, coexistence of diploid and polyploid individuals in sympatric populations is unlikely unless cytotypes are prezygotically isolated through assortative pollination. Here, we investigated the pre-mating barriers involved in the maintenance of three co-occurring cytotypes from the genus Gymnadenia (Orchidaceae): tetraploid and octoploid G. conopsea and tetraploid G. densiflora. We assessed differences in flowering phenology, floral morphology, and visual and olfactory cues, which could lead to assortative mating. Gas chromatography coupled with Electronic supplementary material The online version of this article (electroantennographic detection was used to identify scent compounds with physiological activity in the two main pollinators, Deilephila porcellus and Autographa gamma. The importance of olfactory cues was also assessed in the field by analysing the moths' responses to the olfactory display of the plants, and by following the pollinator's behaviour on artificial arrays. Our complex approach demonstrated that the coexistence of Gymnadenia cytotypes in mixed-ploidy populations was only partly explained by differences in floral phenology, as cytotypes with overlapping flowering (i.e., octoploid G. conopsea and tetraploid G. densiflora) might freely exchange pollen due to only 1 mm differences in spur lengths and the lack of assortative behaviour of pollinators. While floral colour among the cytotypes was similar, floral scent differed significantly. Though both pollinator species seemed to physiologically detect these differences, and the floral scent alone was sufficient to attract them, pollinators did not use this cue to discriminate the cytotypes in the field. The absence of premating barriers among cytotypes, except partial temporal segregation, suggests the existence of other mechanisms involved in the cytotypes' coexistence. The genetic differences in ITS sequences among cytotypes were used to discuss the cytotype's origin.
The challenge of conservation biology is to make models that predict population dynamics and have a high probability of accurately tracking population change (increase, decrease, constancy). In this study we tested whether the transition model is accurate enough to predict population persistence and size 13 years down and whether after 13 years populations had achieved a stable stage distribution. We modeled 6 small populations of an epiphytic orchid using a Lefkovitch type analysis to predict population growth pattern based on monthly surveys for approximately 1.5 years. In addition, sensitivity and elasticity analyses were used to identify life stages with high sensitivity or elasticity that have the largest influence on population growth rate. We re-censused the populations 13 years after the first study and compared the structure of the populations to predictions based on the earlier census data. Effective population growth rates were similar to those expected except for one where the population went extinct. The prediction slightly (but not significantly) overestimated the actual population growth rates of some populations. Elasticity analysis revealed that the adult stage is critical in the life cycle. The observed stage distributions of the populations were not stable at the beginning of the survey and neither were they after 13 years. We suggest that this might be caused by external perturbations that result in unequal mortality between life stages and stochastic recruitment events. The ability of the matrices to predict population size approximately eight generations in the future is encouraging and warrants the continued use of these approaches for PVA.
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