Nectaries are common in Ranunculaceae. These secretory structures, however, have not been studied in detail despite their importance in plant-animal interactions, and data relating to the structure of nectary spurs, which are so characteristic of several genera of this family, remain scarce. In order to redress this imbalance, we sought, in the present paper, to analyze the anatomical and ultrastructural organization of the nectary spurs of four representatives of Ranunculaceae, i.e., Aconitum lycoctonum L., Aquilegia vulgaris L., Consolida regalis Gray, and Delphinium elatum L. Nectary spurs were examined using light, fluorescence, scanning electron, and transmission electron microscopy. The floral nectaries of A. lycoctonum and A. vulgaris are situated at the apices of the spurs, whereas in C. regalis and D. elatum, the nectary is located along the floor surface of the spurs. Nectar in C. regalis and D. elatum is exuded through micro-channels in the cuticle, whereas in A. lycoctonum and A. vulgaris, it is released by means of cell wall disruption, indicating that the method of nectar secretion here is holocrine. Structurally, the nectary of all four investigated species is quite similar, and its cells are typical of nectar-producing cells described in the literature. It is proposed that in A. lycoctonum and A. vulgaris, disruption of the cell wall and the release of the entire cell contents into the spur cavity contribute to the composition of the nectar that the latter contains, enriching it with cytoplasmic components. We conclude that the manner of nectar exudation may vary considerably between closely related plant species, regardless of their geographical origin and phylogeny.

Although originally ascribed to the genus Gymnadenia R. Br. (Orchidinae: Orchidaceae), the Balkan endemic orchid G. frivaldii Hampe ex Griseb. has since been more frequently assigned to Pseudorchis Séguier (syn. Leucorchis E. Mey., Bicchia Parl.). Molecular phylogenetic analysis using the ITS region of rDNA reveals a large disparity between the two genera and demonstrates that frivaldii is embedded well within Gymnadenia s.s. Macromorphological and SEM studies further elucidate the floral and vegetative similarities between G. frivaldii and Pseudorchis, notably the heterochronically reduced gynostemium and small, short‐spurred labellum; these similarities represent convergent evolutionary transitions, whereas other characters such as contrasting stigma and tuber morphologies provide stronger phylogenetic signals. The sequence‐based phylogeny suggests that G. frivaldii represents one of three cases of independent paedomorphic floral reduction inferred in the genus; simplification has been more severe than in G. odoratissima but less severe than in the closely related Gymnadenia subgenus Nigritella. Alternatively, an effectively instantaneous evolutionary origin through hybridisation with a (most likely diploid) species of subgenus Nigritella remains possible. Reports of rare hybridisation between G. frivaldii and members of subgenus Nigritella are acceptably well documented, whereas reports of hybridisation with several other more phylogenetically distant orchid species (including the often sympatric Pseudorchis albida) are considered less secure.

Plant-pollinator interactions are thought to have shaped much of floral evolution. Yet the relative importance of pollinator shifts and coevolutionary interactions for among-population variation in floral traits in animal-pollinated species is poorly known. This study examined the adaptive significance of spur length in the moth-pollinated orchid Platanthera bifolia.Geographical variation in the length of the floral spur of P. bifolia was documented in relation to variation in the pollinator fauna across Scandinavia, and a reciprocal translocation experiment was conducted in south-east Sweden between a long-spurred woodland population and a short-spurred grassland population.Spur length and pollinator fauna varied among regions and habitats, and spur length was positively correlated with the proboscis length of local pollinators. In the reciprocal translocation experiment, long-spurred woodland plants had higher pollination success than short-spurred grassland plants at the woodland site, while no significant difference was observed at the grassland site.The results are consistent with the hypothesis that optimal floral phenotype varies with the morphology of the local pollinators, and that the evolution of spur length in P. bifolia has been largely driven by pollinator shifts.

Spatial variation in plant-pollinator interactions may cause variation in pollinator-mediated selection on floral traits, but to establish this link conclusively experimental studies are needed. We quantified pollinator-mediated selection on flowering phenology and morphology in four populations of the fragrant orchid Gymnadenia conopsea, and compared selection mediated by diurnal and nocturnal pollinators in two of the populations. Variation in pollinator-mediated selection explained most of the among-population variation in the strength of directional and correlational selection. Pollinators mediated correlational selection on pairs of display traits, and on one display trait and spur length, a trait affecting pollination efficiency. Only nocturnal pollinators selected for longer spurs, and mediated stronger selection on the number of flowers compared with diurnal pollinators in one population. The two types of pollinators caused correlational selection on different pairs of traits and selected for different combinations of spur length and number of flowers. The results demonstrate that spatial variation in interactions with pollinators may result in differences in directional and correlational selection on floral traits in a plant with a semi-generalized pollination system, and suggest that differences in the relative importance of diurnal and nocturnal pollinators can cause variation in selection. © 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.

Nectar spurs (tubular outgrowths of a floral organ which contain, or give the appearance of containing, nectar) are hypothesized to be a 'key innovation' which can lead to rapid speciation within a lineage, because they are involved in pollinator specificity. Despite the ecological importance of nectar spurs, relatively little is known about their development. We used a comparative approach to investigate variation in nectar spur length in a clade of eight Iberian toadflaxes.Spur growth was measured at the macroscopic level over time in all eight species, and growth rate and growth duration compared. Evolution of growth rate was reconstructed across the phylogeny. Within the clade we then focused on Linaria becerrae and Linaria clementei, a pair of sister species which have extremely long and short spurs, respectively. Characterization at a micromorphological level was performed across a range of key developmental stages to determine whether the difference in spur length is due to differential cell expansion or cell division.We detected a significant difference in the evolved growth rates, while developmental timing of both the initiation and the end of spur growth remained similar. Cell number is three times higher in the long spurred L. becerrae compared with L. clementei, whereas cell length is only 1.3 times greater. In addition, overall anisotropy of mature cells is not significantly different between the two species.We found that changes in cell number and therefore in cell division largely explain evolution of spur length. This contrasts with previous studies in Aquilegia which have found that variation in nectar spur length is due to directed cell expansion (anisotropy) over variable time frames. Our study adds to knowledge about nectar spur development in a comparative context and indicates that different systems may have evolved nectar spurs using disparate mechanisms.

In petaloid monocots expansion of B-gene expression into whorl 1 of the flower results in two whorls of petaloid organs (tepals), as opposed to sepals in whorl 1 of typical eudicot flowers. Recently, new gene-silencing technologies have provided the first functional data to support this, in the genus Tricyrtis (Liliaceae).Copyright © 2016 Elsevier Ltd. All rights reserved.

Determining the developmental programs underlying morphological variation is key to elucidating the evolutionary processes that generated the stunning biodiversity of the angiosperms. Here, we characterize developmental and transcriptional dynamics of the elaborate petal nectar spur of Aquilegia (columbine) in species with contrasting pollination syndromes and spur morphologies.We collected petal epidermal cell number and length data across four Aquilegia species, two with short, curved nectar spurs of the bee-pollination syndrome, and two with long, straight spurs of the hummingbird syndrome. We also performed RNA-seq on A. brevistyla (bee) and A. canadensis (hummingbird) distal and proximal spur compartments at multiple developmental stages. Finally, we intersected these datasets with a previous QTL mapping study on spur length and shape to identify new candidate loci.The differential growth between the proximal and distal surfaces of curved spurs is primarily driven by differential cell division. However, independent transitions to straight spurs in the hummingbird syndrome have evolved by increasing differential cell elongation between spur surfaces. The RNA-seq data reveal these tissues to be transcriptionally distinct and point to auxin signaling as being involved with the differential cell elongation responsible for the evolution of straight spurs. We identify several promising candidate genes for future study.Our study, taken together with previous work in Aquilegia, reveals the complexity of the developmental mechanisms underlying trait variation in this system. The framework we established here will lead to exciting future work examining candidate genes and processes involved in the rapid radiation of the genus. This article is protected by copyright. All rights reserved.This article is protected by copyright. All rights reserved.

Floral nectar spurs are widely considered a key innovation promoting diversification in angiosperms by means of pollinator shifts. We investigated the macroevolutionary dynamics of nectar spurs in the tribe Antirrhineae (Plantaginaceae), which contains 29 genera and 300-400 species (70-80% spurred). The effect of nectar spurs on diversification was tested, with special focus on Linaria, the genus with the highest number of species. We generated the most comprehensive phylogeny of Antirrhineae to date and reconstructed the evolution of nectar spurs. Diversification rate heterogeneity was investigated using trait-dependent and trait-independent methods, and accounting for taxonomic uncertainty. The association between changes in spur length and speciation was examined within Linaria using model testing and ancestral state reconstructions. We inferred four independent acquisitions of nectar spurs. Diversification analyses revealed that nectar spurs are loosely associated with increased diversification rates. Detected rate shifts were delayed by 5-15 Myr with respect to the acquisition of the trait. Active evolution of spur length, fitting a speciational model, was inferred in Linaria, which is consistent with a scenario of pollinator shifts driving diversification. Nectar spurs played a role in diversification of the Antirrhineae, but diversification dynamics can only be fully explained by the complex interaction of multiple biotic and abiotic factors.© 2018 The Authors. New Phytologist © 2018 New Phytologist Trust.

The plant genus Halenia (Gentianaceae) consists of herbs growing in temperate and tropical alpine habitats and most species possess flowers in which nectar is produced in spurs. This probably helps reward only specialized long-tongued pollinators, and a narrow pollinator/flower relationship is thought to accelerate diversification rates (a key innovation). To test the pattern of diversification of Halenia against the unspurred sister group we reconstructed phylogenetic relationships among 22 species plus outgroups using nuclear ITS and chloroplast rpl16 intron sequence data. We show that Halenia originated in East Asia and migrated via North America into Central America. From there, it colonized South America three times independently, probably within the last million years. Significant changes in diversification rates were found during the evolution of Halenia using a sister group method, a likelihood method, and a diversity-through-time plot. In contrast to other studies, we could not observe a direct speciation rate effect of the evolution of nectar spurs in comparison with the unspurred sister group of Halenia. Rather, increases in diversification occurred following the colonization of Central and South America by spurred progenitor taxa. This later switch in diversification may have resulted from the availability of new geographical and ecological opportunities, or from the availability of more and different pollinators in these regions. Following the latter hypothesis, the nectar spurs were a preadaption and functioned as a key innovation only in this new biotic environment. After an initial rapid increase, a reduction in diversification rate was observed in Central America, probably illustrating density dependence of speciation rates. Finally, we found preliminary evidence for the key innovation hypothesis in geologically young spurred and unspurred lineages of Halenia in South America.

The Orchidaceae have a history of recurring convergent evolution in floral function as nectar production has evolved repeatedly from an ancestral nectarless state. However, orchids exhibit considerable diversity in nectary type, position and morphology, indicating that this convergence arose from alternative adaptive solutions. Using the genus Disa, this study asks whether repeated evolution of floral nectaries involved recapitulation of the same nectary type or diversifying innovation. Epidermis morphology of closely related nectar-producing and nectarless species is also compared in order to identify histological changes that accompanied the gain or loss of nectar production.The micromorphology of nectaries and positionally equivalent tissues in nectarless species was examined with light and scanning electron microscopy. This information was subjected to phylogenetic analyses to reconstruct nectary evolution and compare characteristics of nectar-producing and nectarless species.Two nectary types evolved in Disa. Nectar exudation by modified stomata in floral spurs evolved twice, whereas exudation by a secretory epidermis evolved six times in different perianth segments. The spur epidermis of nectarless species exhibited considerable micromorphological variation, including strongly textured surfaces and non-secreting stomata in some species. Epidermis morphology of nectar-producing species did not differ consistently from that of rewardless species at the magnifications used in this study, suggesting that transitions from rewardlessness to nectar production are not necessarily accompanied by visible morphological changes but only require sub-cellular modification.Independent nectary evolution in Disa involved both repeated recapitulation of secretory epidermis, which is present in the sister genus Brownleea, and innovation of stomatal nectaries. These contrasting nectary types and positional diversity within types imply weak genetic, developmental or physiological constraints in ancestral, nectarless Disa. Such functional convergence generated by morphologically diverse solutions probably also underlies the extensive diversity of nectary types and positions in the Orchidaceae.

One of Verne Grant's lasting contributions to plant evolutionary biology has been the recognition that differences between plants in floral characters can have a dramatic impact on both pollinator visitation and pollen transfer and thus affect reproductive isolation between nascent plant species (collectively, floral isolation). Here we review some of the concepts and findings from Grant's work on floral isolation, particularly with respect to the genus Aquilegia (Ranunculaceae). It has now been over 50 yr since Grant first published on the role of floral isolation on reproductive isolation and speciation in Aquilegia and we compare and contrast his findings with our own work on this genus. We find that the data largely support Grant's findings and that Aquilegia will continue to offer great opportunities to learn about the processes of adaptation and speciation.

Spatiotemporal variation in the interactions among plants and animals is widespread; yet our conceptual and empirical understanding of this variation is limited to a few types of visitors, mainly herbivores, pollinators, seed predators, and seed dispersers. Despite the ubiquity of nectar robbing and the strength of its effects on plant fitness, we know relatively little about the magnitude and intensity of spatial and temporal variation in interactions among plants and nectar robbers. The purpose of the present study was to quantify spatial and temporal variation in the interactions between a nectar-robbing bumblebee and its host plants. In the Rocky Mountains of Colorado, USA, over 7 years, and multiple locations, we measured levels of nectar robbing by the bumblebee Bombus occidentalis and its interactions with four different host plants, Delphinium nuttallianum (Ranunculaceae), Ipomopsis aggregata (Polemoniaceae), Corydalis caseana (Fumariaceae), and Linaria vulgaris (Scrophulariaceae). Significant variation was found in the robbing rates experienced by different species. Within species, there was variation in robbing rates on an annual basis, on a seasonal basis, among different sites, and within sites. This variation may have important consequences with respect to the population dynamics of host plants as well as selection on floral and flowering traits. Electronic supplementary material is available if you access this article at http://dx.doi.org/10.1007/s00442-002-1060-z. On that page (frame on the left side), a link takes you directly to the supplementary materials.

Balsaminaceae consist of two genera, the monospecific Hydrocera and its species-rich sister Impatiens. Although both genera are seemingly rather similar in overall appearance, they differ in ecology, distribution range, habitat preference and morphology. Because morphological support for the current molecular phylogenetic hypothesis of Impatiens is low, a developmental study is necessary in order to obtain better insights into the evolutionary history of the family. Therefore, the floral development of H. triflora and I. omeiana was investigated, representing the most early-diverged lineage of Impatiens, and the observations were compared with the literature.Flowers at all developmental stages were examined using scanning electron microscopy and light microscopy.In Hydrocera, two whorls of five free perianth primordia develop into a less zygomorphic perianth compared with its sister genus. The androecial cap originates from five individual stamen primordia. Post-genital fusion of the upper parts of the filaments result in a filament ring below the anthers. The anthers fuse forming connivent anther-like units. The gynoecium of Hydrocera is pentamerous; it is largely synascidiate in early development. Only then is a symplicate zone formed resulting in style and stigmas. In I. omeiana, the perianth is formed as in Hydrocera. Five individual stamen primordia develop into five stamens, of which the UPPER PART OF THE FILAMENTS CONVERGE WITH EACH OTHER. THE GYNOECIUM OF I. OMEIANA IS TETRAMEROUS; IT APPEARS ANNULAR IN EARLY DEVELOPMENT.Comparison of the present results with developmental data from the literature confirms the perianth morphocline hypothesis in which a congenital fusion of the parts of the perianth results in a shift from pentasepalous to trisepalous flowers. In addition, the development of the androecial cap and the gynoecium follows several distinct ontogenetic sequences within the family.

Exposed nectar presentation is a key trait in flowers specialized for pollination by short-tongued insects. We investigated the pollination of Satyrium microrrhynchum, a rare South African orchid in which nectar is secreted as droplets on long floral hairs ("lollipop hairs") at the mouth of a shallow labellum. Our observations indicate that this orchid is pollinated specifically by two insect species: a cetoniid beetle (Atrichelaphinus tigrina) and a pompilid wasp (Hemipepsis hilaris). Both insects have short mouthparts and remove nectar from the hairs with sweeping motions of their mouthparts. Pollinaria become attached to the upper surface of their heads while they feed on the nectar. Beetles damage the hairs while feeding, which may explain the positive relationship between hair damage and pollination success in plants of S. microrrhynchum from populations where beetles were common. The orchid has cryptic green-yellow flowers with spectral reflectance similar to that of its leaves. The fragrance from plants in three populations, analyzed using gas chromatography coupled to mass spectrometry, was dominated by various terpenoids; linalool was the most abundant. Plants in different populations emitted similar compounds, but eugenol and derivatives of this compound were found in only one of the three populations. In an electrophysiological study (gas chromatography coupled to electroantennography), using antennae of A. tigrina, clear signals were elicited by some of the floral scent compounds.

Field studies in South Africa showed that floral spur length in the Disa draconis complex (Orchidaceae) varies enormously between populations in the southern mountains (means = 32-38 mm), lowland sandplain (mean = 48 mm), and northern mountains (means = 57-72 mm). We tested the hypothesis that divergence in spur length has resulted from selection exerted through pollinator proboscis length. Short-spurred plants in several southern mountain populations, as well as long-spurred plants in one northern mountain population, were pollinated by a horsefly, Philoliche rostrata (Tabanidae), with a proboscis length that varied from 22 to 35 mm among sites. Long-spurred plants on the sandplain were pollinated by the tanglewing fly, Moegistorynchus longirostris (Nemestrinidae), which has a very long proboscis (mean = 57 mm). Selection apparently favors long spurs in sandplain plants, as artificial shortening of spurs resulted in a significant decline in pollen receipt and fruit set, although pollinaria removal was not significantly affected. Fruit set in the study populations was limited by pollen availability, which further suggests that selection on spur length occurs mainly through the female component of reproductive success.© 1997 The Society for the Study of Evolution.

Phylogenetic studies are often hampered by the independent evolution of characters that may potentially obscure relationships. The adaptive significance of the nectar spur and its evolution within the Mexican lobeliads (Campanulaceae) is considered here. The taxonomic delimitations of Heterotoma from the Mexican species within the genera Lobelia and Calcaratolobelia were tested. Independent molecular data were gathered to determine whether the Mexican spurred lobeliads should be treated as distinct genera. The internal transcribed spacer (ITS) region from 18–26S nuclear rDNA and chloroplast DNA from the 3′ trnK intron were sequenced from 14 representative species. Our data suggest that Heterotoma, as originally conceived, is a good evolutionary unit within Lobelia and that the presence of a nectar spur is an important morphological character that can be used in defining phylogenetic position. This study also suggests that morphological changes associated with hummingbird pollination have evolved more than once in the Mexican lobeliads, from small blue‐flowered, insect‐pollinated relatives.

Pollen and nectar are the primary rewards offered by flowers to pollinators. In floral visitors of some plant species, pollen thieves and nectar robbers cause the reduction in pollen grain number and nectar volume, respectively. However, it remains unclear whether the absence of either of the two rewards in a given flower reduces its attraction to nectar- and pollen-collecting pollinators. We hypothesized that flowers removed of either nectar or pollen would attract fewer pollinators. We studied protandrous Impatiens oxyanthera, whose flowers provide bumblebee pollinators with both nectar and pollen in the male phase. We conducted floral reward manipulation experiments to explore how the removal of either nectar or pollen from flowers influences pollinator behaviour by comparing their visitation rates and visit duration. Compared with the control flowers, the flowers removed of pollen attracted significantly more bumblebee pollinators per 30 min, but the flowers removed of nectar or those removed of both pollen and nectar attracted significantly fewer bumblebee pollinators per 30 min. Moreover, the visit duration of bumblebee pollinators to control flowers or flowers removed of pollen was longer than that to flowers removed of nectar or those removed of both pollen and nectar. Our investigations indicated that compared with control flowers, the flowers removed of nectar attracted fewer bumblebee pollinators, supporting our hypothesis. However, our other hypothesis that pollen removal would reduce pollinator visits was not supported by our results. Instead, compared with control flowers, the flowers that contained only nectar attracted more bumblebee pollinators. Nectar seems to be the main reward, and bumblebee pollinators mainly used the absence of pollen as a visual signal to locate I. oxyanthera flowers with a potentially higher amount of nectar.

The native and naturalized species of Fumariaceae in southern Africa are reviewed, and keys and full descriptions are provided. All relevant regional synonyms are given and the indigenous species are illustrated. Three native genera with four species are recognized. The previously taxonomically unplaced genus Cysticapnos Mill. (3 spp.) is included with two other monotypic native South African genera, Discocapnos Cham. Schltdl. and  Trigonocapnos Schltr., in an enlarged circum­scription of subtribe Discocapninae, which is morphologically defined by tendrilliferous leaves and mostly sessile racemes with the lowermost flower basal. Two subspecies are recognized in Cysticapnos vesicaria (E.Mey. ex Bemh.) Lidén, subsp. vesicaria with fewer, smaller flowers and subsp. namaquensis J.C.Manning Goldblatt for plants from Namaqualand with more numerous, often larger flowers. C.  parviflora Lidén appears to be nothing more than a dwarf-fruited form of C. vesicaria, in which heterocarpy has long been known. C. pruinosa (E.Mey. ex Bemh.) Lidén is recorded for the first time to be a short-lived perennial and not an annual, thus unique in the tribe Fumarieae. Discocapnos mundii var.  dregei Harv. from the southern Cape is treated as subsp. dregei (Harv.) J.C.Manning Goldblatt on account of its slightly smaller fruits and significant geographical disjunction from subsp. mundii. Three introduced species are included, Fumaria capreolata L., F. muralis Sond. ex W.D.Koch subsp.  muralis and F. parviflora Lam. var. parviflora, although only the last two appear to be truly naturalized.

Two indicators of a clade's success are its diversity (number of included species) and its disparity (extent of morphospace occupied by its members). Many large genera show high diversity with low disparity, while others such as Euphorbia and Drosophila are highly diverse but also exhibit high disparity. The largest genera are often characterized by key innovations that often, but not necessarily, coincide with their diagnostic apomorphies. In terms of their contribution to speciation, apomorphies are either permissive (e.g. flightlessness) or generative (e.g. nectariferous spurs).Except for Drosophila, virtually no genus among those with the highest diversity or disparity includes species currently studied as model species in developmental genetics or evolutionary developmental biology (evo-devo). An evo-devo approach is, however, potentially important to understand how diversity and disparity could rapidly increase in the largest genera currently accepted by taxonomists. The most promising directions for future research and a set of key questions to be addressed are presented in this review.From an evo-devo perspective, the evolution of clades with high diversity and/or disparity can be addressed from three main perspectives: (1) evolvability, in terms of release from previous constraints and of the presence of genetic or developmental conditions favouring multiple parallel occurrences of a given evolutionary transition and its reversal; (2) phenotypic plasticity as a facilitator of speciation; and (3) modularity, heterochrony and a coupling between the complexity of the life cycle and the evolution of diversity and disparity in a clade. This simple preliminary analysis suggests a set of topics that deserve priority for scrutiny, including the possible role of saltational evolution in the origination of high diversity and/or disparity, the predictability of morphological evolution following release from a former constraint, and the extent and the possible causes of a positive correlation between diversity and disparity and the complexity of the life cycle.© The Author 2015. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

\n The role of petal spurs and specialized pollinator interactions has been studied since Darwin.\n Aquilegia\n petal spurs exhibit striking size and shape diversity, correlated with specialized pollinators ranging from bees to hawkmoths in a textbook example of adaptive radiation. Despite the evolutionary significance of spur length, remarkably little is known about\n Aquilegia\n spur morphogenesis and its evolution. Using experimental measurements, both at tissue and cellular levels, combined with numerical modelling, we have investigated the relative roles of cell divisions and cell shape in determining the morphology of the\n Aquilegia\n petal spur. Contrary to decades-old hypotheses implicating a discrete meristematic zone as the driver of spur growth, we find that\n Aquilegia\n petal spurs develop via anisotropic cell expansion. Furthermore, changes in cell anisotropy account for 99 per cent of the spur-length variation in the genus, suggesting that the true evolutionary innovation underlying the rapid radiation of\n Aquilegia\n was the mechanism of tuning cell shape.\n

Out-of-plane tissue deformations are key morphogenetic events during plant and animal development that generate 3D shapes, such as flowers or limbs. However, the mechanisms by which spatiotemporal patterns of gene expression modify cellular behaviours to generate such deformations remain to be established. We use the Snapdragon flower as a model system to address this problem. Combining cellular analysis with tissue-level modelling, we show that an orthogonal pattern of growth orientations plays a key role in generating out-of-plane deformations. This growth pattern is most likely oriented by a polarity field, highlighted by PIN1 protein localisation, and is modulated by dorsoventral gene activity. The orthogonal growth pattern interacts with other patterns of differential growth to create tissue conflicts that shape the flower. Similar shape changes can be generated by contraction as well as growth, suggesting tissue conflict resolution provides a flexible morphogenetic mechanism for generating shape diversity in plants and animals.

Platanthera ciliaris is a butterfly-pollinated, terrestrial orchid with a loose terminal raceme of 10-50 orange flowers, characterized by a long nectariferous spur. In the southeastern United States, P. ciliaris occurs in the Appalachian mountains and coastal-plain physiographic provinces, but it is found rarely in the intervening Piedmont. In 1983 and 1984, detailed observations of two populations within these disjunct areas revealed that the butterfly species that serve as the primary pollinators differ sharply. In the mountains, Papilio troilus (spicebush swallowtail) was the most frequent and effective visitor, whereas in the coastal plain, P. palamedes (palamedes swallowtail) was the predominant pollinator. Proboscis lengths of P. troilus (mean = 23.3 mm) were significantly shorter than those of P. palamedes (mean = 28.7 mm). Floral characters, most notably spur length, also differed significantly between mountain (mean = 23.8 mm) and coastal-plain (mean = 25.6 mm) plants. In both 1983 and 1984, levels of pollinator service, as assessed by rates of removal and insertion of pollinia, were higher in the mountains (0.81 and 0.86) than in the coastal plain (0.63 and 0.67). In addition, fruit-set was significantly greater in mountain (83.9% in 1983, 86.5% in 1984) than in coastal-plain (63.8% in 1983, 65.5% in 1984) populations. We hypothesize that selection pressure exerted through pollinator proboscis lengths has resulted in pollination ecotypes of P. ciliaris. The short-spurred mountain plants appear to be in equilibrium with their short-tongued butterfly pollinators, receiving high levels of effective pollination and achieving high levels of fruit-set. Coastal-plain plants produce flowers with longer spurs which, nevertheless, are shorter than optimal for insuring that very long-tongued butterflies make contact with their pollinia. Thus, effective pollinator service and fruit-set are reduced. Correlation analyses in 1984 showed a positive relationship between spur length and fruit-set only in the coastal plain. The presence of long-tongued nectar thieves in the coastal plain may also contribute to lower effective pollinator service and fruit-set. Results of reciprocal-transplant studies attempting to determine the genetic basis of floral characters, including spur length, were inconclusive but suggest that differences between mountain and coastal-plain plants are not due solely to phenotypic plasticity.© 1990 The Society for the Study of Evolution.

The anatomy and the ultrastructure of spur nectaries of <em>Cymnadenia conopsea</em> at different developmental stages were investigated. The secretory epidermis surrounded the inside of spur and formed many unicellular papillae, which significantly enlarged the secretory surface. At the activity stage the epidermal cells contained characteristic plastids with well developed intraplastidal membrane system and numerous osmiophillic globules. The contact of plastids and endoplasmic reticulum indicates a possibility of the involvement of these structures in the secretory processes. The cell wall and the cuticle did not form a barrier for the secreted nectar and no pores or cracks were visible in the cuticle covering secretory papillae.

Larger inflorescences in reward-producing plants can benefit plants by increasing both pollinator attraction and the duration of visits by individual pollinators. However, ultimately, inflorescence size is determined by the balance between the benefits of large inflorescences and the increased cost of geitonogamy. At present, little is known about the relationship between inflorescence size and fecundity in deceptive plants. Given that pollinators are likely to leave inflorescences lacking rewards quickly, it seems unlikely that longer pollinator visits and the risk of geitonogamy would be strong selective pressures in these species, which indicates that pollinator attraction might be the most important factor influencing their inflorescence size. Here we examined the pollination ecology of the deceptive orchid Cephalanthera falcata in order to clarify the effects of inflorescence size on the fruit set of this non-rewarding species. Field observations of the floral visitors showed that C. falcata is pollinated by the andrenid bee Andrena aburana, whilst pollination experiments demonstrated that this orchid species is neither autogamous nor apogamous, but is strongly pollinator dependent. Three consecutive years of field observations revealed that fruit set was positively correlated with the number of flowers per inflorescence. These results provide strong evidence that the nectarless orchid C. falcata benefits from producing larger inflorescences that attract a greater number of innate pollinators. Large inflorescences may have a greater positive effect on fruit set in deceptive plants because a growing number of studies suggest that fruit set in reward-producing plants is usually unaffected by display size.

Floral nectar spurs are hypothesized to have had a major role in the evolution of floral diversity and plant-pollinator coadaptation. We examined variation in the degree of nectar spur curvature in two species of jewelweed (Impatiens capensis and Impatiens pallida) pollinated by different sets of pollinators. To distinguish between adaptive and nonadaptive explanations for between-species differences in curvature, we determined the relationship between spur curvature and pollen removal, which is one estimate of male reproductive success. Spur curvature exhibited considerable variation both within and among three populations, with spur angles ranging from 0° to 297°. A greenhouse experiment determined that spur curvature of I. capensis flowers has a broad-sense heritability of 0.636. Laboratory experiments indicated that flowers having recurved spurs deposit significantly more pollen grains on hummingbird visitors than flowers having perpendicular spurs, apparently as a result of greater contact between the androecium of curve-spurred flowers and the upper bill of hummingbirds. We also found a significant relationship between spur curvature and flower length, suggesting a developmental link between the two traits. We discuss the degree of spur curvature in bird-pollinated I. capensis as a function of both adaptive evolution and developmental constraint.Key words: pollination, nectar spurs, hummingbirds, Impatiens capensis, Impatiens pallida, pollen export, floral morphology, floral polymorphisms.

The attractiveness of nectar rewards depends both on the quantity of nectar produced and on its chemical composition. It is known that nectar quantity and chemical composition can differ in plant species depending on the main pollinator associated with the species. The main aims of this study were to test formally whether nectar traits are adapted to pollination syndromes in the speciose Balsaminaceae and, if so, whether a combination of nectar traits mirrors pollination syndromes.Comparative methods based on Ornstein-Uhlenbeck models were used to test whether nectar volume, nectar sucrose proportion, sugar and amino acid concentration and amino acid composition had evolved as a function of pollination syndromes in 57 species of Balsaminaceae. Cluster analysis and ordination were performed to derive clusters of species resembling each other in nectar composition.Evolutionary models for nectar volume and nectar sucrose proportion performed best when including information on pollination syndrome, while including such information improve model fit neither for sugar and amino acid concentration nor for amino acid composition. A significant relationship emerged between pollination syndrome and the combined nectar traits.Our results show that nectar volume and nectar sucrose proportion evolve rapidly towards optimal values associated with different pollination syndromes. The detection of a signal indicating that nectar traits in combination are to a certain extent able to predict pollination syndromes in Balsaminaceae suggests that a holistic approach including the whole set of nectar traits helps us to better understand evolution of nectar composition in response to pollinators.© The Author(s) 2019. Published by Oxford University Press on behalf of the Annals of Botany Company.

Flower specialisation of angiosperms includes the occluded corollas of snapdragons (Antirrhinum and some relatives), which have been postulated to be one of the most efficient structures to physical limit access to pollinators. The Iberian Peninsula harbours the highest number of species (18 Iberian of the 20 species of Antirrhinum) that potentially share similar pollinator fauna. Crossing experiments with 18 Iberian species from this study and literature revealed a general pattern of self-incompatibility (SI) - failure in this SI system has been also observed in a few plants - which indicates the need for pollinator agents in Antirrhinum pollination. Field surveys in natural conditions (304 h) found flower visitation (>85%) almost exclusively by 11 species of bee (Anthophora fulvitarsis, Anthophora plumipes, Anthidium sticticum, Apis mellifera, Bombus hortorum, Bombus pascuorum, Bombus ruderatus, Bombus terrestris, Chalicodoma lefebvrei, Chalicodoma pyrenaica and Xylocopa violacea). This result covering the majority of Antirrhinum species suggests that large bees of the two long-tongued bee families (Megachilidae, Apidae) are the major pollinators of Antirrhinum. A bipartite modularity analysis revealed two pollinator systems of long-tongued bees: (i) the long-studied system of bumblebees (Bombus spp.) associated with nine primarily northern species of Antirrhinum; and (ii) a newly proposed pollinator system involving other large bees associated with seven species primarily distributed in southern Mediterranean areas.© 2017 German Botanical Society and The Royal Botanical Society of the Netherlands.

The structure of the floral nectaries of <i>Lonicera kamtschatica</i> was examined using light microscopy, scanning electron microscopy and transmission electron microscopy. Nectariferous tissues are located in the lower portion of the corolla tube. It was found that the secretory tissue of the nectary was composed of two layers of epidermal formations: short papillae and about 3x longer unicellular trichomes. They cover the adaxial surface of a small spur. Nectar secretion takes place through the apical portion of the trichomes and papillae. The cell wall of the upper part of the trichome has protuberances participating in nectar transfer to the subcuticular space which reaches large dimensions. The lateral walls of the trichomes are saturated with cutin. The papillae have much thicker walls than the trichomes. In the papillae, there are no wall protuberances. Less secretion accumulates in the subcuticular cavities of the papillae than in the trichomes.

The petal spur of the basal eudicot Aquilegia is a key innovation associated with the adaptive radiation of the genus. Previous studies have shown that diversification of Aquilegia spur length can be predominantly attributed to variation in cell elongation. However, the genetic pathways that control the development of petal spurs are still being investigated. Here, we focus on a pair of closely related homologs of the AUXIN RESPONSE FACTOR family, AqARF6 and AqARF8, to explore their roles in Aquileiga coerulea petal spur development. Expression analyses of the two genes show that they are broadly expressed in vegetative and floral organs, but have relatively higher expression in petal spurs, particularly at later stages. Knockdown of the two AqARF6 and AqARF8 transcripts using virus-induced gene silencing resulted in largely petal-specific defects, including a significant reduction in spur length due to a decrease in cell elongation. These spurs also exhibited an absence of nectar production, which was correlated with downregulation of STYLISH homologs that have previously been shown to control nectary development. This study provides the first evidence of ARF6/8 homolog-mediated petal development outside the core eudicots. The genes appear to be specifically required for cell elongation and nectary maturation in the Aquilegia petal spur.© 2020 The Authors. New Phytologist © 2020 New Phytologist Trust.

在动植物的相互关系中，盗蜜行为被认为是一种不同于普通传粉者的非正常访花行为。动物之所以要采取这种特殊的觅食策略，有假说认为是由访花者的口器和植物的花部形态不匹配造成的，也有认为是盗蜜行为提高了觅食效率从而使盗蜜者受益。在盗蜜现象中， 盗蜜者和宿主植物之间的关系是复杂的。盗蜜对宿主植物的影响尤其是对其繁殖适合度的影响归纳起来有正面、负面以及中性3类。与此同时，盗蜜者的种类, 性别及其掠食行为差异不仅与生境因素密切相关，而且会对宿主植物的繁殖成功产生直接或间接的影响。另外，盗蜜者的存在无疑对其它正常传粉者的访花行为也产生一定的影响，从而间接地影响宿主植物的繁殖成功, 而植物在花部形态上也出现了对盗蜜现象的适应性进化。作者认为, 盗蜜是短嘴蜂对长管型花最有效的一种掠食策略, 它不仅增加了盗蜜者对资源的利用能力, 而且由于盗蜜对宿主植物繁殖成功的不同的影响使其具有调节盗蜜者和宿主之间种群动态的作用, 两者的彼此适应是一种协同进化的结果。

Urophysa is an Asian endemic genus in the Ranunculaceae, but data on floral organogenesis, which would be a useful complement to molecular data in clarifying the relationship with closely related taxa (Aquilegia and Semiaquilegia) in Ranunculaceae, are completely lacking. We used scanning electron microscopy and light microscopy to study the floral development of Urophysa rockii Ulbrich, a recently rediscovered species in this genus. The sepals are initiated spirally, whereas other organs are nonsimultaneously whorled; the floral phyllotaxis is whorled. Primordia of the sepals are lunular and truncate, but those of the petals and stamens are hemispherical and rounded. After sepal initiation, there is a delay in development, but the initiation of petals and stamens is continuous. The developmental sequence of the microspores in the stamens is centrifugal, although the stamens are initiated centripetally. The early developmental stages of the staminodes are similar to those of the stamens, although much smaller, so they may be phylogenetically homologous organs. The carpel primordia are lunular in shape and plicate. The mature ovule is anatropous and bitegmic. Urophysa shows similar floral development features as Aquilegia and Semiaquilegia, although with some differences, which supports the relationship inferred by DNA sequence data.