Eco-evolutionary processes shaping floral nectar sugar composition

We experimentally tested the hypothesis that the extensive within-plant variation of nectar sugar composition in Helleborus foetidus (Ranunculaceae) and other species results from differences between flowers and nectaries in pollinator visitation history. Experiments were conducted to mimic single-nectary visits by wild-caught individuals of the main bee pollinators of H. foetidus, which were assayed for their capacity to modify the sugar composition of natural and artificial nectar. Experimental nectar probing with bee mouthparts induced extensive changes in proportional sugar composition 48 h after treatment, and bee taxa differed widely in their effects. Nectar probing by Andrena, medium-sized Anthophoridae, Apis mellifera, and Lasioglossum had no subsequent effects on nectar sugar composition, while probing by Bombus terrestris and B. pratorum induced an extensive reduction in percentage sucrose, a marked increase in percentage fructose, and a slight increase in percentage glucose. Results support the hypothesis that stochastic variations among flowers or nectaries in the taxonomic identity of recent visitors and their relative visitation frequencies may eventually generate very small-scale mosaics in nectar sugar composition. Changes in nectar sugar composition following bumblebee probing may be the consequence of nectar contamination with pollinator-borne nectarivorous yeasts.

A first comprehensive dataset of nectar sugar composition and concentration in Bromeliaceae is presented, covering 111 species belonging to all three subfamilies. Based on this dataset, we examined the relationship between nectar traits and pollination syndromes in the family. Sugars in samples were assayed by high pressure liquid chromatography. All sampled species were grouped into three broad categories (trochilophilous, chiropterophilous, or lepidopterophilous) according to their main pollination mode. Significant differences between the different pollination syndromes were found in nectar sugar composition as well as concentration. For a total of four genera (Guzmania, Pitcairnia, Tillandsia and Vriesea), a comparison of nectar composition showed significant differences between trochilophilous and chiropterophilous species. Data presented here indicate that the characteristics of nectar in Bromeliaceae are predominantly determined by putative adaptations of nectar sugars to preferences of the pollinators rather than by phylogenetic relations.

ABSTRACTSeveral characteristics of flowers and fruits have been suggested as comprising syndromes of characters that indicate particular classes of pollinators and fruit dispersers. Common phylogenetic history among species, however, may also significantly influence these characters and obscure or enhance perceived patterns of plant syndromes. We analyzed the proportions of glucose, fructose, and sucrose by paper chromatography in the nectar and fruit juice of 525 tropical and subtropical plant species to test whether sugar chemistry was correlated with volant vertebrate pollinator or fruit disperser classes. Samples were taken from Old World and New World species and the calculations kept separate. Kruskal‐Wallis tests of family means showed significant deviations in the percent sucrose content among pollinatorl disperser classes. Mann‐WhitneyU‐tests showed significant differences among nectars of all pollinator classes but fruit juices differed only due to the high sucrose content of megachiropteran dispersed fruits. In addition, sign tests of samples occurring within families showed significant correlations between percentage sucrose content and pollinator/disperser classes. Passerine nectars had low sucrose content. In striking contrast, the nectar of hummingbird flowers had very high sucrose content. The Microchiroptera nectars showed hexose richness with a sucrose content somewhat greater than that of passerine flowers. Megachiroptera flowers showed sucrose‐rich nectars. The results for fruits were comparable to those for nectars. Passerine fruits were hexose dominated, microchiropteran fruits had a sucrose content similar to passerine fruits, and megachiropteran fruits were sucrose‐rich. We speculate on the evolutionary sequence of changes in nectar and fruit juice sugar composition and suggest that future investigations consider the chemistry of other food sources such as pollen and leaves. Only with these additions and other ecological studies can the full interplay of such plant‐animal interactions be anticipated.

Background and AimsFloral nectar can be variable in composition, influencing pollinator behaviour and the composition of honey derived from it. The non-peroxide antibacterial activity of mānuka (Leptospermum scoparium, Myrtaceae) honey results from the chemical conversion of the triose sugar dihydroxyacetone (DHA), after DHA accumulates for an unknown reason in the nectar. This study examined variation in nectar DHA, glucose, fructose and sucrose content with floral stage of development, between mānuka genotypes with differing flower morphology, and in response to water stress.MethodsSix mānuka genotypes were grown without nectar-feeding insects. Stages of flower development were defined, nectar was harvested and its composition was compared between stages and genotypes, and with floral morphology. Water stress was imposed and its effect on nectar composition was examined.Key ResultsNectar was present from soon after flower opening until the end of petal abscission, with the quantity of accumulated nectar sugars rising, then stabilizing or falling, indicating nectar secretion followed by reabsorption in some genotypes. The quantity of DHA, the ratio of DHA to other nectar sugars and the fructose to glucose ratio also varied with stage of development, indicating differences in rates of production and reabsorption between nectar components. Nectar composition and yield per flower also differed between genotypes, although neither was positively related to nectary area or stomatal density. Drying soil had no effect on nectar composition or yield, but variation in nectar yield was correlated with temperature prior to nectar sampling.ConclusionsMānuka nectar yield and composition are strongly influenced by plant genotype, flower age and the environment. There were clear stoichiometric relationships between glucose, fructose and sucrose per flower, but DHA per flower was only weakly correlated with the amount of other sugars, suggesting that accumulation of the triose sugar is indirectly coupled to secretion of the larger sugars by the nectary parenchyma.

Many features of flowers comprise the key elements of the pollinating strategies of flowering plants. Our aims were to describe features that attract pollinators and to identify the pollination system of Petunia interior, a species for which bees have been suggested as the probable pollinators. Therefore, we described the morphology and floral biology, assessed nectar production, concentration, and composition, examined reproductive mode and identified pollinators. P. interior has a purple, infundibuliform, zygomorphic corolla with a short and wide tube and blue pollen. Flower opening and pollen release were asynchronous throughout the day. The pollen grains have pollenkitt on the surface. The nectar sugar composition has a proportion of sucrose lower than the proportion of glucose + fructose, and the nectar supply was constant, in small amounts, at a concentration between 16.6-23.1 %. The reproductive system is xenogamous and bees were the exclusive pollinators. P. interior exhibits a set of floral traits that prevent self-pollination and maintains attractiveness to the bees. The greater reproductive success under natural conditions highlights the importance of bees for the reproductive success of P. interior. As far as floral traits are concerned, only the sugar concentration in the nectar does not correspond to melittophily.

The nectar-sugar composition of 40 species of purportedly hummingbird- pollinated plants in 12 families was determined by high-performance liquid chromatography. The distribution of relative percentages by mass for the major sugars--fructose, glucose, and sucrose--was normal about their means. The means and 95% confidence limits were: fructose, 15% (3%-33%); glucose, 11% (0%-23%); and sucrose, 74% (55%-94%). One sample also contained a trace of maltose. Salvia spathacea contained only fructose and sucrose. The data, primarily from southwestern North America, define a range of nectar composition, possibly representing the preferences of hummingbirds. These data support assertions of adaptive convergence in the sugar composition of nectar in hummingbird-pollinated species.

The large terrestrial orchid genus Satyrium underwent evolutionary radiations in the Cape floral region and the grasslands of southern and eastern Africa. These radiations were accompanied by tremendous diversification of the unusual twin-spurred flowers that characterize the genus, but pollination data required to interpret these patterns of floral evolution have been lacking for grassland species in the genus. Here we document pollinators, nectar properties, and levels of pollination success for 11 grassland Satyrium species in southern and south-central Africa. Pollinators of these species include bees, beetles, butterflies, hawkmoths, noctuid moths, long-proboscid flies, and sunbirds. Most species appear to be specialized for pollination by one functional pollinator group. Long-proboscid fly pollination systems are reported for the first time in Satyrium (in S. macrophyllum and a high-altitude form of S. neglectum). Floral morphology, especially spur length and rostellum structure, differs markedly among plants with different pollinators, while nectar volume, concentration, and sugar composition are fairly uniform across species. Most taxa exhibited high levels of pollination success (>50% of flowers pollinated), a trend that can be attributed to the presence of nectar in the twin spurs.

Floral nectar composition has been explained as an adaptation to factors that are either directly or indirectly related to pollinator attraction. However, it is often unclear whether the sugar composition is a direct adaptation to pollinator preferences. Firstly, the lower osmolality of sucrose solutions means that they evaporate more rapidly than hexose solutions, which might be one reason why sucrose-rich nectar is typically found in flowers with long tubes (adapted to long-tongued pollinators), where it is better protected from evaporation than in open or short-tubed flowers. Secondly, it can be assumed that temperature-dependent evaporation is generally lower during the night than during the day so that selection pressure to secrete nectar with high osmolality (i.e. hexose-rich solutions) is relaxed for night-active flowers pollinated at night. Thirdly, the breeding system may affect selection pressure on nectar traits; that is, for pollinator-independent, self-pollinated plants, a lower selective pressure on nectar traits can be assumed, leading to a higher variability of nectar sugar composition independent of pollinator preferences, nectar accessibility and nectar protection. To analyse the relations between flower tube length, day vs. night pollination and self-pollination, the nectar sugar composition was investigated in 78 European Caryophylloideae (Caryophyllaceae) with different pollination modes (diurnal, nocturnal, self-pollination) using high-performance liquid chromatography (HPLC). All Caryophylleae species (Dianthus and relatives) were found to have nectar with more than 50% sucrose, whereas the sugar composition of Sileneae species (Silene and relatives) ranged from 0% to 98.2%. In the genus Silene, a clear dichotomous distribution of sucrose- and hexose-dominant nectars is evident. We found a positive correlation between the flower tube length and sucrose content in Caryophylloideae, particularly in day-flowering species, using both conventional analyses and phylogenetically independent contrasts.

The genetic basis of host plant use by phytophagous insects can provide insight into the evolution of ecological niches, especially phenomena such as specialization and phylogenetic conservatism. We carried out a quantitative genetic analysis of multiple host use traits, estimated on five species of host plants, in the Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae). Mean values of all characters varied among host plants, providing evidence that adaptation to plants may require evolution of both behavioral (preference) and post-ingestive physiological (performance) characteristics. Significant additive genetic variation was detected for several characters on several hosts, but not in the capacity to use the two major hosts, a pattern that might be caused by directional selection. No negative genetic correlations across hosts were detected for any 'performance' traits, i.e. we found no evidence of trade-offs in fitness on different plants. Larval consumption was positively genetically correlated across host plants, suggesting that diet generalization might evolve as a distinct trait, rather than by independent evolution of feeding responses to each plant species, but several other traits did not show this pattern. We explored genetic correlations among traits expressed on a given plant species, in a first effort to shed light on the number of independent traits that may evolve in response to selection for host-plant utilization. Most traits were not correlated with each other, implying that adaptation to a novel potential host could be a complex, multidimensional 'character' that might constrain adaptation and contribute to the pronounced ecological specialization and the phylogenetic niche conservatism that characterize many clades of phytophagous insects.

Flower morphology, phenology and visitor patterns in an alpine community on Mt Olympos, Greece

Variation in floral morphology and timing of bloom are common among cultivars of northern highbush blueberry (Vaccinium corymbosum L.). These differences can affect the ability of commercial pollinators to effectively pollinate and fertilize ovules, which can impact fruit set, berry size, and ultimate yields. New northern highbush blueberry cultivars may exhibit differences in flower morphology, which could impact pollination and fruit set. Evaluation of new cultivars compared to standard industry cultivars is of specific importance in predicting potential pollination constraints by honey bee (Apis mellifera), one of the primary pollinators in North America, and in developing optimal hive stocking densities. Three new cultivars (‘Blue Ribbon’, ‘Top Shelf’, and ‘Cargo’) were compared to an industry standard (‘Duke’) for floral morphology and relative bloom phenology. All new cultivars had significantly smaller flowers as compared to ‘Duke’; however, ‘Top Shelf’ flowers exhibited a unique characteristic whereby many flowers (81%) had partially fused petals. Reduced flower size of new cultivars suggests floral morphology may be a constraint to effective pollination by honey bees and/or these cultivars will require higher honey bee stocking densities for effective pollination. However, the partially unfused petals in ‘Top Shelf’ may provide improved exposure to the stamen and pistil, which could increase accessibility to pollinators like honey bee and promote pollination and fruit set.

Flowering phenology is central to plant reproductive success and can relate to morphological traits such as size and quality of flowers, but phenology–trait associations of flowers remain unclear. Phenology–trait associations can vary within and among species, especially across climate gradients, due to allocational trade‐offs in response to selective pressures from abiotic stress and/or biotic factors such as pollinator availability. It is unknown whether floral morphology and phenology experience coordinated selection across climate gradients or vary independently. Understanding the drivers of trait variation and its implications for floral phenology may be especially relevant in mountains where climate and abiotic gradients change sharply with elevation. At higher elevations, increasing cold and aridity may favour higher quality, stress‐tolerant flowers that withstand harsher conditions better than cheaper flowers, but trade‐off with size and showiness to pollinators. Relatively benign conditions at lower elevations may permit more flexible phenotypes and weaker association between trait and phenology by allowing species more leeway for flowering times compared with the short growing season at higher elevations. We examined trait–phenology associations within and among 250 herbaceous species in the Western Himalayas, spanning 2000–4500 m AMSL. Intraspecific variation in traits and phenology decreased from low to high elevation at the community scale, but species showed idiosyncratic responses. Trait–trait correlations indicated stronger selection and constraints on phenotypes and phenology at higher elevations. However, trait–phenology correlations were relatively weak throughout, suggesting independent selection on flower morphology vs. phenology. Overall, climate imposed strong selection on flower traits and regulated floral phenology, but without any strong or consistent associations between traits and phenology. It is likely that flower morphology responded to multiple abiotic and biotic drivers, within and across elevations, whereas phenology mainly responded to temperature and thaw. Our findings suggest that selection from ongoing warming will operate separately on floral morphology and phenology of alpine plant communities, and mechanisms and consequences should be evaluated using experiments. Read the free Plain Language Summary for this article on the Journal blog.

The number of exotic plant species that have been introduced into the United States far exceeds that of other groups of organisms, and many of these have become invasive. As in many regions of the globe, invasive members of the thistle tribe, Cardueae, are highly problematic in the California Floristic Province, an established biodiversity hotspot. While Darwin's naturalization hypothesis posits that plant invaders closely related to native species would be at a disadvantage, evidence has been found that introduced thistles more closely related to native species are more likely to become invasive. To elucidate the mechanisms behind this pattern, we modelled the ecological niches of thistle species present in the province and compared niche similarity between taxa and their evolutionary relatedness, using fossil-calibrated molecular phylogenies of the tribe. The predicted niches of invasive species were found to have higher degrees of overlap with native species than noninvasive introduced species do, and pairwise niche distance was significantly correlated with phylogenetic distance, suggesting phylogenetic niche conservatism. Invasive thistles also displayed superior dispersal capabilities compared to noninvasive introduced species, and these capabilities exhibited a phylogenetic signal. By analysing the modelled ecological niches and dispersal capabilities of over a hundred thistle species, we demonstrate that exapted preferences to the invaded environment may explain why close exotic relatives may make bad neighbours in the thistle tribe.

The purpose of our study was to determine the role of flower structure and nectar composition in shaping the reproductive success (RS) of the generalist orchid Epipactis helleborine in natural and anthropogenic populations. We supposed that the distinct character of two groups of habitats creates different conditions for plant-pollinator relationships, thus influencing reproductive success in E. helleborine populations. Both pollinaria removal (PR) and fruiting (FRS) were differentiated between the populations. On average, FRS was almost two times higher in the anthropogenic than in the natural populations. The difference between the two population groups in PR was smaller but still statistically significant. RS parameters were correlated with some floral display and flower traits. Floral display influenced RS only in three anthropogenic populations. Flower traits had a weak influence on RS (10 of the 192 cases analyzed). The more important trait in shaping RS was nectar chemistry. The nectar of E. helleborine is relatively diluted with a lower sugar concentration in the anthropogenic than in the natural populations. In the natural populations, domination of sucrose over hexoses was found, while in the anthropogenic populations, hexoses were more abundant and the participation of sugars was balanced. In some populations, sugars influenced RS. In E. helleborine nectar, 20 proteogenic and 7 non-proteogenic amino acids (AAs) were found with a clear domination of glutamic acid. We noted relationships between some AAs and RS, but distinct AAs shaped RS in different populations, and their impact was independent of their participation. Our results indicate that the flower structure and nectar composition of E. helleborine reflect its generalistic character and meet the requirements of a wide range of pollinators. Simultaneously, the differentiation of flower traits suggests a variation in pollinator assemblages in particular populations. Knowledge about the factors influencing RS in distinct habitats helps to understand the evolutionary potential of species and to understand mechanisms and processes crucial for shaping interactions between plants and pollinators.

To understand the relationships between floral morphologies and the reproductive strategies of plants, the contributions of multiple pollinators to plant reproductive success need to be examined. To outline recent advances on this topic, we review studies of five bumblebee‐pollinated plants in Japan. These plant species are pollinated by one to three bumblebee species differing in proboscis length: Bombus consobrinus, Bombus diversus and Bombus honshuensis. These pollination systems allow us to compare the reproductive strategies of plants pollinated by single and multiple bumblebee species. First, we address the effects of morphological correspondence between flowers and pollinators on the reproductive success of plants. In plants with narrow tubular flowers, Isodon umbrosus and Clematis stans, pollination effectiveness depended on the morphological correspondence between floral tube length and pollinator mouthparts. These flowers are subject to varied directions and intensities of selection by multiple bumblebee species, resulting in the maintenance of large variation in floral morphology within and between populations, and even within individuals. In contrast, pollination effectiveness in plants having flowers with a wide entrance, Melampyrum roseum and Hosta sieboldiana, did not differ among bumblebee species because of good correspondence between floral morphology and bumblebee body size, resulting in small morphological variations among flowers. Thus, generalized pollination systems with multiple pollinator species favor floral traits that enable the use of various pollinators. Second, we address the quantity and quality of the pollination service in bumblebee‐pollinated plants. Isodon umbrosus, with its generalized pollination system, suffered from pollen limitation because of low pollination quality. In contrast, Isodon effusus, which has a specialized pollination system, suffered from pollen limitation because of low pollination quantity. Isodon umbrosus produced large numbers of short tubular flowers to improve pollination quantity, whereas I. effusus produced small numbers of long tubular flowers to improve pollination quality, resulting in a clear trade‐off between the size and number of flowers within plants. In M. roseum, which has a generalized pollination system, three bumblebee species achieved sufficient pollination, but seed production was limited by an increased rate of self‐pollination. This is because this species invests more resources in cross‐pollinated seeds by selective abortion of selfed seeds. Our results demonstrate that the strategy of generalized plants is to improve pollination quantity, whereas the strategy of specialized plants is to improve pollination quality.