Evidence for progenitor-derivative speciation in sexually deceptive orchids

Background and Aims Sexually deceptive orchids of the genus Ophrys use mimicry of pollinator females to attract specific pollinators. Pollinator shifts may drive speciation in Ophrys, since novel pollinators may in principle act as isolating factors immediately. It is thus possible that evolution of novel species occurs rapidly and with a progenitor-derivative pattern. The aims of this study are to compare genetic structure and diversity among widespread and geographically restricted Ophrys taxa, to test whether genetic structure is associated with specific pollinators, and to investigate whether any widespread species may have acted as a progenitor for the evolution of more restricted taxa. Methods Genetic differentiation and diversity were investigated in O. leucadica and O. cinereophila, the two taxa of the Ophrys fusca sensu lato complex widespread in the Aegean, and three geographically restricted taxa from Rhodes, O. attaviria, O. parvula and O. persephonae, all differing in their specific pollinators. This was done using amplified fragment length polymorphism (AFLP) DNA fingerprinting, and sequencing of the low-copy nuclear gene LEAFY (LFY). Key Results All taxa were found to be separate genetic entities, with O. leucadica forming two geographic groups from the west and east of the Aegean. Genetic structure was significantly shaped by pollinators and geography, and comparison of sequence and AFLP data revealed ancestral polymorphisms shared among several taxa. Among the sampled taxa, O. leucadica harbours the greatest genetic differentiation and geographic structure, and the highest genetic diversity. Part of the genome of O. parvula, endemic to Rhodes, may be derived from O. leucadica. Conclusions Pollinators probably influence the genetic structure of the investigated Ophrys species. The genetic pattern identified is consistent with O. leucadica being the oldest of the sampled taxa, making O. leucadica a candidate progenitor species from which more restricted taxa such as O. parvula may have evolve

Floral visual signal increases reproductive success in a sexually deceptive orchid

Sexually deceptive orchids mimic signals emitted by female insects in order to attract mate-searching males. Specific attraction of the targeted pollinator is achieved by sex pheromone mimicry, which constitutes the major attraction channel. In close vicinity of the flower, visual signals may enhance attraction, as was shown recently in the sexually deceptive orchid Ophrys heldreichii. Here, we conducted an in situ manipulation experiment in two populations of O. heldreichii on Crete to investigate whether the presence/absence of the conspicuous pink perianth affects reproductive success in two natural orchid populations. We estimated reproductive success of three treatment groups (with intact, removed and artificial perianth) throughout the flowering period as pollinaria removal (male reproductive success) and massulae deposition (female reproductive success). Reproductive success was significantly increased by the presence of a strong visual signal—the conspicuous perianth—in one study population, however, not in the second, most likely due to the low pollinator abundance in the latter population. This study provides further evidence that the coloured perianth in O. heldreichii is adaptive and thus adds to the olfactory signal to maximise pollinator attraction and reproductive success

Speciation, pattern recognition and the maximization of pollination: general questions and answers given by the reproductive biology of the orchid genus Ophrys

Pollination syndromes evolved under the reciprocal selection of pollinators and plants (coevolution). Here, the two main methods are reviewed which are applied to prove such selection. (i) The indirect method is a cross-lineage approach using phylogenetical trees to understand the phylogeny. Thus, features of single origin can be distinguished from those with multiple origins. Nearly all pollination modes originate in multiple evolutionary ways. (ii) The most frequent pollinators cause the strongest selection because they are responsible for the plant’s most successful reproduction. The European sexually deceptive orchid genus Ophrys provides an example of a more direct way to prove selection because the attraction of a pollinator is species specific. Most members of the genus have remarkably variable flowers. The variability of the signals given off by the flowers enables the deceived pollinator males to learn individual flower patterns. They thus avoid already visited Ophrys flowers, interpreting them as females rejecting them. As the males will not return to these individually recognizable flowers, the pollinators´ learning behavior causes cross-pollination and prevents the orchid’s self-pollination.© The Author(s) 201

Scent variation and hybridization cause the displacement of a sexually deceptive orchid species

In the sexually deceptive orchid genus Ophrys, reproductive isolation is based on the specific attraction of males of a single pollinator species, mostly bees, by mimicking the female sex pheromone of this species. Changes in the floral odor can lead to hybridization, introgression, and possibly speciation. We investigated hybrid swarms of O. lupercalis and O. iricolor on Sardinia using behavioral, electrophysiological (GC-EAD), chemical, morphological, and genetic methods (AFLPs). In behavioral experiments, approximately 20% of the flowers from both species and hybrids were attractive to the "wrong" or both pollinator species. Analysis of the EAD-active hydrocarbons in the floral odor showed an overlap in the two species, whereby hybrid individuals could not be separated from O. iricolor. The genetic analysis confirmed the hybridization of the species. Plants of O. iricolor and hybrids are genetically indistinguishable and form an O. iricolor x lupercalis hybrid population. Remaining plants of O. lupercalis will possibly be displaced by the O. iricolor x lupercalis hybrid population in the future. Our study showed that in deceptive orchids, variation in the pollinator attracting cues, in this case, scent, can be the first step for speciation and at the same time cause the displacement of a species

Fast Fourier Transform (FFT) of stimuli used in the honeybee experiments.

<p>Whereas the two types of crosses (A) differ widely in their FFT, the <i>Ophrys</i> pattern of same inflorescences (B) as well as those from different plants (C) show high similarity in their FFTs.</p

Degree of similarity among patterns from three <i>O</i>. <i>heldreichii</i> populations.

<p>Each box consists of 15 comparisons of randomly chosen flowers of the same (dark grey) or from different inflorescences (light grey). Four different features were compared: (A) pattern overlap, (B) pattern symmetry, (C) relative pattern surface area, and (D) contour density. Boxes represent inter-quartile range with median values. Stars indicate statistical differences after Bonferroni correction (***, p<0.00025; **, p<0.0025; *, p<0.0125; n.s., not significant).</p

Duration of approach, copulation and post-copulation scanning behavior of <i>Eucera berlandi</i> males on an <i>Ophrys heldreichii</i> flower.

<p>Boxes represent inter-quartile range with median values, whiskers represent the 5 and 95 percentiles.</p