Flower morphology evolution in Jaborosa (Solanaceae): shape and size variation associated with contrasting pollination modes.

Flower morphology is considered an important factor in species diversification because it may influence the efficiency of pollination in different ways (e.g. attraction and mechanical fit with different groups of pollinators). In the present study, we quantified the variation in flower morphology (i.e. shape and size) of the diverse South American genus Jaborosa Juss. (Solanaceae) in relation to contrasting pollination modes: rewarding pollination either by moths or by generalist small insects versus brood-site deceptive pollination by saprophilous flies. We examined variations of flower morphology in frontal (pollinator attraction) and sagittal (functional fit with pollinators) views in 12 Jaborosa species using geometric morphometric methods and comparative approaches to infer whether flower shape evolution, not attributable to flower allometry or phylogenetic relationship, is associated with shifts in pollination modes. We found remarkable variation in flower morphology among both Jaborosa species and pollination modes, largely in sagittal view. Evolutionary trends in shape of fly-pollinated flowers were mainly attributable to changes in developmental trajectories. Variation in flower architecture facilitated differential pollen placement - on the proboscis of moths, and either on the back or ventral region of saprophilous flies - promoting diversification of the genus. Diversification of shape, independent of size, in most of the studied Jaborosa species would indicate adaptation to contrasting pollination modes.

Floral colour variation of Nicotiana glauca in native and non-native ranges: Testing the role of pollinators' perception and abiotic factors.

Invasive plants displaying disparate pollination environments and abiotic conditions in native and non-native ranges provide ideal systems to test the role of different ecological factors driving flower colour variation. We quantified corolla reflectance of the ornithophilous South American Nicotiana glauca in native populations, where plants are pollinated by hummingbirds, and in populations from two invaded regions: South Africa, where plants are pollinated by sunbirds, and the Balearic island of Mallorca, where plants reproduce by selfing. Using visual modelling we examined how corolla reflectance could be perceived by floral visitors present in each region. Through Mantel tests we assessed a possible association between flower colour and different abiotic factors. Corolla reflectance variation (mainly along medium to long wavelengths, i.e. human green-yellow to red colours) was greater among studied regions than within them. Flower colour was more similar between South America and South Africa, which share birds as pollinators. Within invaded regions, corolla reflectance variation was lower in South Africa, where populations could not be distinguished from each other by sunbirds, than in Spain, where populations could be distinguished from each other by their occasional visitors. Differences in corolla colour among populations were partially associated with differences in temperature. Our findings suggest that shifts in flower colour of N. glauca across native and invaded ranges could be shaped by changes in both pollination environment and climatic factors. This is the first study on plant invasions considering visual perception of different pollinators and abiotic drivers of flower colour variation.

The least effective pollinator principle: specialized morphology despite generalized ecology.

The large body of work on the adaptation of plants to pollinators is still somewhat incomplete because most studies focus on one-to-one interactions. How will adaptation proceed in a multi-pollinator environment? According to Stebbins' Most Effective Pollinator Principle, 'the characteristics of the flower will be moulded by those pollinators that visit it most frequently and effectively.' To test this hypothesis, we studied the pollination biology of Pelargonium incrassatum (Geraniaceae) in the Namaqualand Region of Southern Africa. This species has a long floral tube and we expected its most important pollinator to have a long proboscis. Contrary to expectations, the most important pollinator was a short proboscid fly (a new species of Prosoeca), while Prosoeca peringueyi, which had a proboscis that matched the floral tube length, was a rare visitor. Consistent with the high degree of trait mismatching, we did not detect selection on tube length at most sites. The paradox of mismatching traits can be resolved by considering the strength of the trade-off involved. Adaptation to the rare species can apparently occur without incurring the cost of reduced pollination by the abundant species. Generally, species may often evolve specialized morphology if they do not incur the cost of ecological specialization.

Drift effects on the multivariate floral phenotype of Calceolaria polyrhiza during a post-glacial expansion in Patagonia.

Quaternary environmental changes substantially impacted the landscape and promoted rapid evolutionary changes in many species; however, analyses of adaptive phenotypic variation in plants have usually neglected the underlying historical context. Here, we associate phylogeography and phenotypic evolution by analysing the divergence of Calceolaria polyrhiza multivariate floral phenotype after a Pleistocene post-glacial expansion in Patagonia. Phenotypic matrix (P) properties (size, shape, orientation and phenotypic integration) of six refugium and six recent populations from two different phylogroups were compared following different approaches. We found that P-matrix shape and orientation remained stable despite the strong phylogeographic footprint of post-glacial expansion. However, average proportional reductions in matrix size supported the expectation that drift had a significant effect on the floral phenotype in the northern phylogroup. When phylogeographic history was not included in the analyses, the results overestimated phenotypic differences, whereas under explicit phylogeographic control, drift appeared as the best explanation for matrix differences. In general, recent populations showed a larger phenotypic divergence among them, but a lower overall phenotypic variation than refugium populations. Random Skewers analyses indicated a lower potential response to selection in recently colonized populations than in refugium populations. We discuss that the combination of phylogeographic analyses with geographical distribution of functional phenotypic (genotypic) variation is critical not only to understand how historical effects influence adaptive evolution, but also to improve field comparisons in evolutionary ecology studies.

The search for Pleiades in trait constellations: functional integration and phenotypic selection in the complex flowers of Morrenia brachystephana (Apocynaceae).

Pollinator-mediated natural selection on single traits, such as corolla tube or spur length, has been well documented. However, flower phenotypes are usually complex, and selection is expected to act on several traits that functionally interact rather than on a single isolated trait. Despite the fact that selection on complex phenotypes is expectedly widespread, multivariate selection modelling on such phenotypes still remains under-explored in plants. Species of the subfamily Asclepiadoideae (Apocynaceae) provide an opportunity to study such complex flower contrivances integrated by fine-scaled organs from disparate developmental origin. We studied the correlation structure among linear floral traits (i) by testing a priori morphological, functional or developmental hypotheses among traits and (ii) by exploring the organization of flower covariation, considering alternative expectations of modular organization or whole flower integration through conditional dependence analysis (CDA) and integration matrices. The phenotypic selection approach was applied to determine whether floral traits involved in the functioning of the pollination mechanism were affected by natural selection. Floral integration was low, suggesting that flowers are organized in more than just one correlation pleiad; our hypothetical functional correlation matrix was significantly correlated with the empirical matrix, and the CDA revealed three putative modules. Analyses of phenotypic selection showed significant linear and correlational gradients, lending support to expectations of functional interactions between floral traits. Significant correlational selection gradients found involved traits of different floral whorls, providing evidence for the existence of functional integration across developmental domains.

Functional morphology and wasp pollination of two South American asclepiads (Asclepiadoideae-Apocynaceae).

UNLABELLED: BACKGROUND AND AIMS The extreme complexity of asclepiad flowers (Asclepiadoideae-Apocynaceae) has generated particular interest in the pollination biology of this group of plants especially in the mechanisms involved in the pollination processes. This study compares two South American species, Morrenia odorata and Morrenia brachystephana, with respect to morphology and anatomy of flower structures, dynamic aspects of the pollination mechanism, diversity of visitors and effectiveness of pollinators. METHODS: Floral structure was studied with fresh and fixed flowers following classical techniques. The pollination mechanism was studied by visiting fresh flowers in the laboratory with artificial pollinator body parts created with an eyelash. Morphometric and nectar measurements were also taken. Pollen transfer efficiency in the flowers was calculated by recording the frequency of removed and inserted pollinia. Visitor activity was recorded in the field, and floral visitors were captured for subsequent analysis of pollen loads. Finally, pollinator effectiveness was calculated with an index. KEY RESULTS: The detailed structure of the flowers revealed a complex system of guide rails and chambers precisely arranged in order to achieve effective pollinaria transport. Morrenia odorata is functionally specialized for wasp pollination, and M. brachystephana for wasp and bee pollination. Pollinators transport chains of pollinaria adhered to their mouthparts. CONCLUSIONS: Morrenia odorata and M. brachystephana present differences in the morphology and size of their corona, gynostegium and pollinaria, which explain the differences in details of the functioning of the general pollination mechanism. Pollination is performed by different groups of highly effective pollinators. Morrenia species are specialized for pollination mainly by several species of wasps, a specialized pollination which has been poorly studied. In particular, pompilid wasps are reported as important pollinators in other regions outside South Africa. A putative new function of nectar in asclepiads is presented, as it would be contributing to the pollination mechanism.

A simple floral fragrance and unusual osmophore structure in Cyclopogon elatus (Orchidaceae).

We studied gland morphology, anatomy and the chemical composition of the floral fragrance in the sweat bee-pollinated orchid Cyclopogon elatus. This is apparently the first such analysis for any Cyclopogon species, and one of very few studies in which both odour and osmophore are characterised in a nectar-rewarding orchid. Structures responsible for floral scent production were localised with neutral red staining and histochemical assays for lipids and starch. Their morphology and anatomy were studied with scanning electron microscopy and light microscopy thin sections, respectively. Fragrance samples were collected using SPME fibres and analysed with GC-MS. Anatomical evidence suggests that two parallel oval-shaped patches of unicellular trichomes on the abaxial surface of the labellum are osmophores. These are rich in stored lipids, while the parenchyma surrounding the vascular bundles contains starch. Only freshly opened flowers produced odours, while buds and withered flowers lacked scent. The chemical composition of the odour was dominated (>99.8%) by a single compound, trans-4,8-dimethyl-nona-1,3,7-triene (DMNT). Gland anatomy and position on the outside of the perianth are unusual for scent glands in general. The presence of DMNT, a nearly ubiquitous compound in herbivore-induced vegetative emissions and one of the major floral volatiles of Yucca, is not surprising in view of hypotheses on the evolutionary origin of flower scents, suggesting that wound volatiles are utilised as kairomonal attractants by florivores whose activities result in pollination.