What are the drivers of female success in food-deceptive orchids?

A large suite of floral signals, and environmental and biotic characteristics influence the behavior of pollinators, affecting the female success of food-deceptive orchids. In this study, we examined the many factors shaping the reproductive output of three orchid taxa: Dactylorhiza majalis, D. incarnata var. incarnata, and D. fuchsii. We applied a statistical model to correlate female success (number of fruit sets) with individual characteristics (plant and inflorescence height, number of flowers, and spur length), number of pollinaria removed, flowering time, and density of floral units of co-flowering rewarding plants. Our findings suggested that the broad spectrum of variations in Dactylorhiza's morphological traits, floral display, and flowering phenology within different environmental contexts has a significant impact on their reproductive success. The number of fruits increased with an increase in the number of pollinaria removed in the studied Dactylorhiza taxa. In contrast, a higher number of flowers per inflorescence and higher inflorescences in relation to individual height always decreased fruit set. We observed that low number of co-flowering rewarding plants in populations could affect the Dactylorhiza reproductive output as magnets and competitor plants. The synchronization of flowering, or lack thereof, between Dactylorhiza and rewarding plants can limit reproductive success. This demonstrates that the food deception strategy is multidirectional, and reproductive output can vary considerably both spatially and temporally within the context of this strategy.

The patterns of inbreeding depression in food-deceptive Dactylorhiza orchids.

Introduction: Inbreeding depression (ID) in food-deceptive plants have been reported previously, however, it has not been often proven that selfed seeds germinate better than outbred ones or that selfing affects ID. To resolved these issues, food-deceptive related Dactylorhiza majalis, D. incarnata var. incarnata and D. fuchsii orchids were investigated. Methods: Hand pollination treatments and control pollination were conducted. Fruit set, number of seeds per fruit, seed length, number of well-developed seeds per fruit, and proportion of in vitro asymbiotic germination seeds, were analyzed in relation to inflorescence levels and used as fitness indicators for these orchids. The ID and pollen limitation were measured. Results: The lowest ID (δ = -1.000) was in D. majalis, and present in combination with a high pollen limitation in its populations. D. fuchsii showed higher ID (δ = 0.366), and D. incarnata var. incarnata weak ID (δ = 0.065), although ID varied between its fitness components. The seed number per fruit differed significantly between the treatments and the inflorescence levels in treatments. Discussion: This study emphasizes that the breeding system rather than the flower position on the inflorescence shaped the quality and quantity of reproductive output. The ID and its effect on germination of food-deceptive orchid seeds undoubtedly played an important role.

Pollen transfer and patterns of reproductive success in pure and mixed populations of nectariferous Platanthera bifolia and P. chlorantha (Orchidaceae).

Plant species evolution is driven by many factors that have different roles in space and time. Using different field and laboratory methods, we studied reproductive patterns and their determinants in pure and mixed P. bifolia and P. chlorantha populations in different habitats. We also considered the probability of hybridisation between these two species and the role of intra-population processes in maintaining species integrity. Generally, we found a high level of reproductive success in both Platantherans. In both species, male (MRS) and female (FRS) reproductive success depended on floral display, and male reproductive success additionally on population structure. The flower traits were only weakly related to reproductive success. Moths' assemblages varied spatially and temporally, and their diversity and numbers were correlated with MRS in the year, when their abundance was markedly lower. Analysis of patterns of pollen transfer showed that pollen was transported up to 25 m (average 8.2 ± 4.83 m) and showed gene exchange between these two Platanthera species. The germination level of both species was significantly lower than seed viability, although P. bifolia seed germinated with higher frequency than P. chlorantha seeds. We noted differences in viability and germination of seeds developed as an effect of experimental interspecies crossings and those developed from natural pollination. The presence of intermediate ecotypes together with observations of spontaneous interspecies crosses in the field and viability of seeds produced in interspecies crossing suggest that both pre- and postzygotic reproductive barriers are not complete and do not prevent hybrid production.

Spatiotemporal Variations in Seed Set and Pollen Limitation in Populations of the Rare Generalist Species Polemonium caeruleum in Poland.

A vast majority of angiosperms are pollinated by animals, and a decline in the number and diversity of insects often affects plant reproduction through pollen limitation. This phenomenon may be particularly severe in rare plant species, whose populations are shrinking. Here, we examined the variability in factors shaping reproductive success and pollen limitation in red-listed Polemonium caeruleum L. During a 5-year study in several populations of P. caeruleum (7-15, depending on year), we assessed the degree of pollen limitation based on differences in seed set between open-pollinated (control) and hand-pollinated flowers. We analysed the effects of flower visitors, population size, and meteorological data on plant reproductive success and pollen limitation. Our study showed that pollen limitation rarely affected P. caeruleum populations, and was present mainly in small populations. Pollen limitation index was negatively affected by the size of population, visitation frequency of all insects, and when considering the visitation frequency of individual groups, also by honeybee visits. Seed production in control treatment was positively influenced by the population size, average monthly precipitation in June and visits of hoverflies, while visits of honeybees, average monthly temperature in September, and average monthly precipitation in August influenced seed production negatively. As generalist plant P. caeruleum can be pollinated by diverse insect groups, however, in small populations their main visitors, the honeybees and bumblebees, may be less attracted, eventually leading to the disappearance of these populations. In pollination of P. caeruleum managed honeybees may play a dual role: while they are the most frequent and efficient flower visitors, their presence decreases seed set in open-pollinated flowers, which is most probably related to efficient pollen collection by these insects.

Deceptive strategy in Dactylorhiza orchids: multidirectional evolution of floral chemistry.

BACKGROUND AND AIMS: The deception strategies of orchids remain poorly understood, especially in regard to the chemical compounds emitted from their flowers and their interaction with various taxonomic groups of pollinators. We investigated the phylogenetic relationships and compared the variation of floral chemical compounds between food-deceptive Dactylorhiza taxa (D. incarnata var. incarnata and D. incarnata var. ochroleuca, D. fuchsii and D. majalis) from populations in north-eastern Poland. We propose a model of the evolution of deception based on floral chemical signals in this genus. METHODS: A Bayesian approach based on polymorphic plastid DNA (trnL, trnF and psbC-trnK), internal transcribed spacer (ITS) sequences and flow cytometry data was applied to confirm the taxonomic status of the studied orchids. We also identified and classified the pollinators and flower visitors in each Dactylorhiza population to the taxonomic level and compared our results with literature data. The chemical composition of pentane and diethyl ether extracts from the flowers was analysed by gas chromatography-mass spectrometry. Variation of the floral chemical components was visualized by non-metric multidimensional scaling based on Bray-Curtis dissimilarity. KEY RESULTS: The genetic distinctiveness of D. incarnata, D. fuchsii and D. majalis was confirmed. No hybrids between them were found, but the chloroplast DNA (cpDNA), ITS haplotypes and flow cytometry showed genetic similarity between D. incarnata var. incarnata and D. incarnata var. ochroleuca. We determined that Apis mellifera (Hymenoptera) was the only shared pollinator of these taxa. Strangalia attenuata and Alosterna tabacicolor (Coleoptera) and Volucella pellucens and V. bombylans (Hymenoptera) were observed pollinating D. fuchsii. Visualization of the emission rates of the 61 floral chemical compounds detected from pentane extracts (mainly hydrocarbons and aldehydes) and the 51 from diethyl extracts (with abundant groups of benzenoids and non-aromatic acids) strongly differentiated D. incarnata, D. fuchsii and D. majalis, while those of the two varieties of D. incarnata (var. incarnata and var. ochroleuca) were almost identical. CONCLUSIONS: While the genetic data clearly supported the distinct lineages of D. incarnata, D. fuchsii and D. majalis, the patterns of emission of their flower chemical compounds were more complex within the series of shared compounds (alkanes and aldehydes) and taxon-specific compounds (benzenoids and esters). Their floral bouquet can influence the sexual, social and feeding behaviour of pollinators in different ways. We observed that the floral chemical compounds attracted both shared and species-specific pollinators to Dactylorhiza, confirming the multidirectional character of floral chemical signals in these food-deceptive taxa. Reduction of species-specific pollination levels in Dactylorhiza orchid taxa may promote hybridization between them.