Seed dormancy release of Halenia elliptica in response to stratification temperature, duration and soil moisture content.

BACKGROUND: Although the effect of cold stratification on seed dormancy release has been extensively studied for many species, knowledge of the role of stratifying temperature, soil moisture content and duration of stratification on seed dormancy release at the population level is limited. Here, we aimed to determine the response of seed dormancy release to these factors in six populations of Halenia elliptica. RESULTS: Seed dormancy release was more responsive to low than high temperatures, and no dormancy break occurred at 8 °C. Seed germination percentage increased first and then remained unchanged as stratifying soil moisture content increased from 0 to 24%. Seed dormancy release of populations from low altitude was more sensitive to increased stratifying temperature and decreased soil moisture content than those from high altitudes. CONCLUSIONS: Temperature and soil moisture changes resulting from global warming could affect seed dormancy release and consequently seedling establishment. Thus, incorporating data on seed dormancy release involving temperature, soil moisture content and stratification duration is beneficial for predicting plant species regeneration, migration and coexistence in a scenario of climate change.

Context-dependency of resource allocation trade-offs highlights constraints to the evolution of floral longevity in a monocarpic herb.

Floral longevity is a critical component of floral display, yet there is a conspicuous paucity of empirical research on its evolution within species. Evolutionary models of floral longevity are grounded in resource allocation theory and propose that selection acts on heritable variation to optimize longevity in light of competing floral construction and maintenance costs. Key assumptions remain untested within wild species. We measured maximum floral longevity alongside protandry, flower size, flower number and flowering rate across families of the monocarpic herb Sabatia angularis grown under high and low resources. We evaluated genetic variation, plasticity and correlations between display traits, including fundamental resource-allocation trade-offs and their interactions with resource availability. All display traits showed significant genetic variation. Resource availability influenced mean floral longevity and flower number, with genetic variation in these responses. Importantly, both floral longevity-flower number and flower number-size trade-offs were significant and stronger under low resources. This study reinforces the application of resource allocation theory to floral display trait evolution. Our work highlights the context-dependency of trade-offs and the potential importance of plasticity in resource allocation, with plants investing in the construction of new flowers at faster rates when resources are high rather than in the maintenance of longer-lived flowers.

Correlation between the timing of autonomous selfing and floral traits: a comparative study from three selfing Gentianopsis species (Gentianaceae).

About 20% of angiosperms employ self-fertilization as their main mating strategy. In this study, we aimed to examine how the selfing timing correlated with floral traits in three Gentianopsis species in which autonomous selfing is achieved through filament elongation. Although the three Gentianopsis species exhibit no significant variation in their capacity for autonomous selfing, flowers of G. grandis last longer, are larger and have a higher corolla biomass, P/O ratios and male biomass allocation than those of G. paludosa, and especially those of G. contorta. Autonomous selfing occurs in the early floral life of G. paludosa and G. contorta and in the later floral life of G. grandis. Seed production mainly results from autonomous selfing in G. paludosa and G. contorta; however, G. grandis could be more described as having a mixed mating system. We suggest that autonomous selfing in later floral life increases the chance of cross-pollination prior to this, while autonomous selfing in early floral life offers a selective advantage to plants by reducing the resource investment in traits that may increase pollinator attraction and visitation.

Small and surrounded: population size and land use intensity interact to determine reliance on autonomous selfing in a monocarpic plant.

Background and Aims: Habitat fragmentation has transformed landscapes globally, leaving remnants embedded within a complex matrix that is rapidly becoming more developed. For many plant populations, the associated factors of decreased size and intensification of land use surrounding them are expected to increase pollen limitation ('PL'), unless autonomous self-pollination provides reproductive assurance ('RA'). Decreased pollinator visitation is often assumed to drive these patterns, but other, less studied mechanisms might include increased heterospecific pollen transfer or decreased conspecific pollen availability via florivory. I investigate how PL and RA and their potential underlying mechanisms vary with population size and land use intensity surrounding populations in the biennial Sabatia angularis (Gentianaceae). Methods: I estimated the capacity for seed production via autonomous self-pollination (i.e. autofertility). Over 2 years in 22 S. angularis populations across a fragmented landscape, I performed emasculation and pollen supplementation experiments measuring RA and PL, and quantified visitation rates of potential pollinators and a pollen consumer, conspecific pollen loads and rates of heterospecific pollen deposition. Key results: Autofertility based on fruit mass was 93 % under PL but only 51.6 % relative to maximal conditions. PL and RA were significant on average across populations in the first year of study. Variation in RA was significantly influenced by the interaction between population size and land use intensity, which in turn rendered PL independent of these factors. Visitation and heterospecific pollen deposition rates were greatest in small populations and declined with population size, while conspecific pollen loads were greatest in intermediate sized populations. Conclusions: Increased reliance on RA is predicted in small S. angularis populations surrounded by intense development, which can explain elevated selfing rates in fragmented populations of plant species more generally. Results from this study point toward forces such as heterospecific pollen transfer, self-pollen limitation or resource availability influencing the need and ability to rely on RA.

Hollow Internodes Permit a Neotropical Understory Plant to Shelter Multiple Mutualistic Ant Species, Obtaining Protection and Nutrient Provisioning (Myrmecotrophy).

The Neotropical understory plant Tachia guianensis (Gentianaceae)-known to shelter the colonies of several ant species in its hollow trunks and branches-does not provide them with food rewards (e.g., extrafloral nectar). We tested whether these ants are opportunistic nesters or whether mutualistic relationships exist as for myrmecophytes or plants sheltering ant colonies in specialized hollow structures in exchange for protection from enemies and/or nutrient provisioning (myrmecotrophy). We noted 37 ant species sheltering inside T. guianensis internodes, three of them accounting for 43.5% of the cases. They protect their host plants from leaf-cutting ant defoliation and termite damage because individuals devoid of associated ants suffered significantly more attacks. Using the stable isotope 15N, we experimentally showed that the tested ant species furnish their host plants with nutrients. Therefore, a mutualism exists. However, because it is associated with numerous ant species, T. guianensis can be considered a nonspecialized myrmecophyte.

Phenotypic selection varies with pollination intensity across populations of Sabatia angularis.

Pollinators are considered primary selective agents acting on plant traits, and thus variation in the strength of the plant-pollinator interaction might drive variation in the opportunity for selection and selection intensity across plant populations. Here, we examine whether these critical evolutionary parameters covary with pollination intensity across wild populations of the biennial Sabatia angularis. We quantified pollination intensity in each of nine S. angularis populations as mean stigmatic pollen load per population. For female fitness and three components, fruit number, fruit set (proportion of flowers setting fruit) and number of seeds per fruit, we evaluated whether the opportunity for selection varied with pollination intensity. We used phenotypic selection analyses to test for interactions between pollination intensity and selection gradients for five floral traits, including flowering phenology. The opportunity for selection via fruit set and seeds per fruit declined significantly with increasing pollen receipt, as expected. We demonstrated significant directional selection on multiple traits across populations. We also found that selection intensity for all traits depended on pollination intensity. Consistent with general theory about the relationship between biotic interaction strength and the intensity of selection, our study suggests that variation in pollination intensity drives variation in selection across S. angularis populations.

Plasticity of floral longevity and floral display in the self-compatible biennial Sabatia angularis (Gentianaceae): untangling the role of multiple components of pollination.

BACKGROUND AND AIMS: Plasticity of floral traits in response to pollination can enable plants to maximize opportunities for pollen import and export under poor pollination conditions, while minimizing costs under favourable ones. Both floral longevity and display are key traits influencing pollination. While pollination-induced flower wilting is widely documented, we lack an understanding of the multifactorial complexity of this response, including the influence of other pollination components, costs of extended longevity and subsequent impacts on floral display. METHODS: Plasticity of floral longevity was experimentally evaluated in Sabatia angularis in response to multiple pollination factors: pollen addition, removal, and source (self, single-donor outcross, multiple-donor outcross) and timing of pollination. Effects of pollen quantity were further evaluated by exploiting variation in autonomous self-pollen deposition. Delayed pollination costs were tested comparing seed set from early versus late pollinations. Finally, I compared floral display metrics (peak floral display, time to peak flower, flowering duration, mean flowering rate) between experimentally pollinated and control plants. KEY RESULTS: Floral longevity was highly plastic in response to pollen addition and its timing, and the response was dose-dependent but insensitive to pollen source. Pollen removal tended to extend floral longevity, but only insofar as it precluded pollination-induced wilting via autonomous self-pollination. Under delayed pollination, the wilting response was faster and no cost was detected. Pollination further led to reduced peak floral displays and condensed flowering periods. CONCLUSIONS: Floral longevity and display plasticity could optimize fitness in S. angularis, a species prone to pollen limitation and high inbreeding depression. Under pollinator scarcity, extended floral longevities offer greater opportunities for pollen receipt and export at no cost to seed set, reproductive assurance via autonomous self-pollination and larger, more attractive floral displays. Under high pollinator availability, shortened longevities lead to smaller displays that should lower the risk of geitonogamy.

Key innovations and climatic niche divergence as drivers of diversification in subtropical Gentianinae in southeastern and eastern Asia.

PREMISE OF THE STUDY: Geological and climatic changes associated with the uplift of the Qinghai-Tibet Plateau (QTP) have been suggested as drivers for biological diversification locally and in neighboring regions. To test this hypothesis, we investigated the niche evolution of Tripterospermum (Gentianaceae) and related Asian genera through time. METHODS: We conducted Species Distribution Modeling using Maximum Entropy Modeling (MaxEnt). Furthermore, we performed stochastic character mapping and produced disparity-through-time plots, and examined putative key innovations using the binary state speciation and extinction approach (BISSE). KEY RESULTS: Kuepferia and Sinogentiana prefer the coolest and driest habitat, having rather conserved niches. Despite a tendency for niche evolution, Crawfurdia and Metagentiana are probably restricted to a narrow distribution range because of their poor dispersal ability. In contrast, Tripterospermum has the broadest niche and occurs under the warmest and wettest conditions. A higher degree of niche evolution and a more efficient dispersal mechanism allowed this genus to diversify more and occupy a broader distribution range. CONCLUSIONS: The QTP genera producing dry capsules, whether displaying niche conservatism (Kuepferia and Sinogentiana) or a tendency for niche evolution (Crawfurdia and Metagentiana), are less species-rich and have a more restricted distribution than Tripterospermum (stronger niche evolution and berry-like fruits). The evolution of berry-like fruits corresponds to increased speciation rates, and could therefore be viewed as a key innovation. In contrast to the majority of studies on plants occurring around the QTP, we find that speciation was probably mediated by niche breadth and dispersal ability rather than geophysical changes.

Apple latent spherical virus vector-induced flowering for shortening the juvenile phase in Japanese gentian and lisianthus plants.

MAIN CONCLUSION: Infection by apple latent spherical virus (ALSV) vectors that promote the expression of Arabidopsis thaliana FLOWERING LOCUS T ( AtFT ) or Gentiana triflora GtFT s accelerates flowering in gentian and lisianthus plants. Apple latent spherical virus (ALSV) has isometric virus particles (25 nm in diameter) that contain two ssRNA species (RNA1 and RNA2) and three capsid proteins (Vp25, Vp20, and Vp24). ALSV vectors are used for foreign gene expression and virus-induced gene silencing in a broad range of plant species. Here, we report the infection by ALSV vectors that express FLOWERING LOCUS T (AtFT) from Arabidopsis thaliana or its homolog GtFT1 from Gentiana triflora in three gentian cultivars ('Iwate Yume Aoi' [early flowering], 'Iwate' [medium flowering], and 'Alta' [late flowering]), and two lisianthus cultivars ('Newlination Pink ver. 2' and 'Torukogikyou daburu mikkusu') promotes flowering within 90 days post-inoculation using particle bombardment. Additionally, seedlings from the progeny of the early-flowering plants were tested by tissue blot hybridization, and the results showed that ALSV was not transmitted to the next generation. The promotion of flowering in the family Gentianaceae by ALSV vectors shortened the juvenile phase from 1-3 years to 3-5 months, and thus, it could be considered as a new plant breeding technique in ornamental gentian and lisianthus plants.

Florivory Modulates the Seed Number-Seed Weight Relationship in Halenia elliptica (Gentianaceae).

Generally, plant reproductive success might be affected negatively by florivory, and the effects may vary depending on the timing and intensity of florivory. To clarify the impacts of florivory by the sawfly larvae (Tenthredinidae) on seed production of Halenia elliptica D. Don, we simulated florivory by removing different proportion of flowers at three reproductive stages in this alpine herb and then examined the seed number per fruit, the seed weight, and the seed mass per fruit of the remaining flowers. Seed number per fruit reduced significantly when flowers were removed at flowering and fruiting stages or when 15% and 60% of flowers were removed. However, seed weight increased significantly after flowers were removed, independent of treatments of reproductive stage and proportion. There was a similar seed mass per fruit between the plants subjected to simulation of florivory and control. The results indicated that florivory modulated the seed number-seed weight relationship in this alpine species. Our study suggested that selective seed abortion and resource reallocation within fruits may ensure fewer but larger seeds, which were expected to be adaptive in the harsh environments.

Back to Gondwanaland: can ancient vicariance explain (some) Indian Ocean disjunct plant distributions?

Oceans, or other wide expanses of inhospitable environment, interrupt present day distributions of many plant groups. Using molecular dating techniques, generally incorporating fossil evidence, we can estimate when such distributions originated. Numerous dating analyses have recently precipitated a paradigm shift in the general explanations for the phenomenon, away from older geological causes, such as continental drift, in favour of more recent, long-distance dispersal (LDD). For example, the 'Gondwanan vicariance' scenario has been dismissed in various studies of Indian Ocean disjunct distributions. We used the gentian tribe Exaceae to reassess this scenario using molecular dating with minimum (fossil), maximum (geological), secondary (from wider analyses) and hypothesis-driven age constraints. Our results indicate that ancient vicariance cannot be ruled out as an explanation for the early origins of Exaceae across Africa, Madagascar and the Indian subcontinent unless a strong assumption is made about the maximum age of Gentianales. However, both the Gondwanan scenario and the available evidence suggest that there were also several, more recent, intercontinental dispersals during the diversification of the group.

Diplostigmaty in plants: a novel mechanism that provides reproductive assurance.

Differentiation of female sexual organs in flowering plants is rare and contrasts with the wide range of male reproductive strategies. An unusual example involves diplostigmaty, the possession of spatially and temporally distinct stigmas in Sebaea (Gentianaceae). Here, the single pistil within a flower has an apical stigma, as occurs in most flowering plants, but also a secondary stigma that occurs midway down the style, which is physically discrete and receptive several days after the apical stigma. We examined the function of diplostigmaty in Sebaea aurea, an insect-pollinated species of the Western Cape of South Africa. Floral manipulations and measurements of fertility and mating patterns provided evidence that basal stigmas function to enable autonomous delayed self-pollination, without limiting opportunities for outcrossing and thus avoiding the costs of seed discounting. We suggest that delayed selfing serves as a mechanism of reproductive assurance in populations with low plant density. The possession of dimorphic stigma function provides a novel example of a flexible mixed-mating strategy in plants that is responsive to changing demographic conditions.

Variation and evolution of herkogamy in Exochaenium (Gentianaceae): implications for the evolution of distyly.

BACKGROUNDS AND AIMS: The spatial separation of stigmas and anthers (herkogamy) in flowering plants functions to reduce self-pollination and avoid interference between pollen dispersal and receipt. Little is known about the evolutionary relationships among the three main forms of herkogamy - approach, reverse and reciprocal herkogamy (distyly) - or about transitions to and from a non-herkogamous condition. This problem was examined in Exochaenium (Gentianaceae), a genus of African herbs that exhibits considerable variation in floral morphology, including the three forms of herkogamy. METHODS: Using maximum parsimony and maximum likelihood methods, the evolutionary history of herkogamic and non-herkogamic conditions was reconstructed from a molecular phylogeny of 15 species of Exochaenium and four outgroup taxa, based on three chloroplast regions, the nuclear ribosomal internal transcribed spacer (ITS1 and 2) and the 5·8S gene. Ancestral character states were determined and the reconstructions were used to evaluate competing models for the origin of reciprocal herkogamy. KEY RESULTS: Reciprocal herkogamy originated once in Exochaenium from an ancestor with approach herkogamy. Reverse herkogamy and the non-herkogamic condition homostyly were derived from heterostyly. Distylous species possessed pendent, slightly zygomorphic flowers, and the single transition to reverse herkogamy was associated with the hawkmoth pollination syndrome. Reductions in flower size characterized three of four independent transitions from reciprocal herkogamy to homostyly. CONCLUSIONS: The results support Lloyd and Webb's model in which distyly originated from an ancestor with approach herkogamy. They also demonstrate the lability of sex organ deployment and implicate pollinators, or their absence, as playing an important role in driving transitions among herkogamic and non-herkogamic conditions.

Pharmacognosy of Enicostemma littorale: a review.

Traditional medicine system of India comprised varieties of plants which are playing a significant role in curing diseases from ancient times. Among them, Enicostemma littorale blume (E. littorale) a perennial herb of the family Gentianaceae is cosmopolitan in occurrence in India. The bittery natured plant acts as a laxative, helps in curing fever, rheumatism, skin diseases, abdominal disorders, snake bite, obesity and helps to regulate blood sugar levels. The plant constituents have been reported for possessing antimicrobial, antioxidant, antiulcer, antiinflammatory, hypolipidaemic, hepatoprotective and hypoglycemic properties. This review provides a bird's eye view about geographical distribution, physicochemical parameters, phytoconstituents and pharmacological properties of E. littorale.

Differences in dichogamy and herkogamy contribute to higher selfing in contrasting environments in the annual Blackstonia perfoliata (Gentianaceae).

BACKGROUND AND AIMS: The establishment of plant populations in novel environments may generate pronounced shifts in floral traits and plant mating systems, particularly when pollinators are scarce. In this study, floral morphology and mating system functioning are compared between recently established and older populations of the annual plant Blackstonia perfoliata that occur in different pollinator environments. METHODS: Hand-pollination and emasculation experiments were conducted to assess the extent of pollinator-mediated pollen deposition and pollen limitation, and the contribution of autonomous selfing to total seed production. Detailed measurements of key floral traits were performed to compare the flower morphology and mating system functioning between plants from both pollination environments. KEY RESULTS: Pollinator-mediated pollen deposition was about twice as low in the recently colonized and pollinator-poor environment compared with the old and pollinator-rich sites, but total seed set was little affected by any type of pollen limitation. The contribution of autonomous selfing to total seed production was higher in the pollinator-poor sites than in the pollinator-rich sites (index of reproductive assurance = 0·56 and 0·17, respectively), and seed production was only poorly affected by selfing, whereas in the pollinator-rich populations selfing reduced total reproductive output by about 40 % compared with outcross pollination. Plants originating from pollinator-poor environments produced smaller flowers that showed significantly lower levels of dichogamy (i.e. protogyny) and herkogamy. These reductions resulted in a 2-fold higher capacity for autonomous selfing under pollinator-free conditions (index of autonomous selfing = 0·81 and 0·41 in plants originating from the pollinator-poor and pollinator-rich environment, respectively). CONCLUSIONS: The results illustrate that plant populations colonizing novel environments can differ markedly in floral morphology and mating system functioning. Due to a temporal shift in the male phase, the breeding system of B. perfoliata shifted from delayed selfing under pollinator-rich conditions towards competing selfing in recently established populations, providing greater reproductive assurance when pollinators and/or reproductive partners are limited.

Sugars and acid invertase mediate the physiological response of Schenkia spicata root cultures to salt stress.

A heterotrophic model system was established in our studies in order to differentiate the effect of high salt concentrations in external medium on growth and sugar metabolism in roots from the effect of reduced sugar availability resulting from decreased photosynthesis under salinity. Soluble sugar content and the activity of acid invertase in root cultures of salt-tolerant (ST) and salt-sensitive (SS) Schenkia spicata (L.) Mansion genotypes were investigated during exposure to different NaCl concentrations (0-200 mM). Their response to severe salinity was characterized by a metabolic adjustment that led to the accumulation of sucrose (Suc) in root tissues. There was clear evidence that cell wall invertase (CW-Inv) is the major contributor to the Suc/hexose ratio in roots during exposure to elevated salinity. The results of CW-Inv activity and immunodetection assays in our study suggest that the regulation of CW-Inv expression is most likely achieved in a salt stress dependent manner. Also, NaCl modulated soluble acid invertase (SA-Inv) expression differentially in SS and ST genotypes of S. spicata. Regardless of the salt treatment, genotype, or the amount of enzyme, SA-Inv activity was generally low, indicating regulation at the posttranslational level. The results suggest no direct role of SA-Inv in the regulation of the root tissue carbohydrate pool and therefore in the control of the availability of glucose and fructose for the primary metabolism and/or osmotic adjustment in the present heterotrophic model system.

Selective seed abortion induced by nectar robbing in the selfing plant Comastoma pulmonarium.

• Self-pollination often provides plants with the benefit of reproductive assurance; thus, it is generally assumed that species' interactions that alter floral attractiveness or rewards, such as nectar robbing, will have little effect on the seed production of selfing species. We challenge this view with experimental data from Comastoma pulmonarium, a selfing annual experiencing a high ratio of nectar robbing in the Qinghai-Tibet Plateau. • We manipulated robbing (robbed or netted) and pollination mode (hand-selfed or hand-outcrossed) in a factorial design and measured the number of developing ovules and mature seeds, together with seed weight and seed germination, in each treatment. • Robbing decreased the number of mature seeds, but not the number of developing ovules, suggesting a negative influence of robbers through indirect effects via selective seed abortion. We found no evidence for early-acting inbreeding depression, but found later-acting inbreeding depression. Our data also suggested that later-acting inbreeding depression of progeny from robbed flowers could be reduced in comparison with that from unrobbed flowers. • We suggest that nectar robbing can have both negative and positive effects on the quantity and quality, respectively, of progeny produced in selfing plants, and challenge the view that robbing has no effect on selfing species.

Inherited biotic protection in a neotropical pioneer plant.

Chelonanthus alatus is a bat-pollinated, pioneer Gentianaceae that clusters in patches where still-standing, dried-out stems are interspersed among live individuals. Flowers bear circum-floral nectaries (CFNs) that are attractive to ants, and seed dispersal is both barochorous and anemochorous. Although, in this study, live individuals never sheltered ant colonies, dried-out hollow stems--that can remain standing for 2 years--did. Workers from species nesting in dried-out stems as well as from ground-nesting species exploited the CFNs of live C. alatus individuals in the same patches during the daytime, but were absent at night (when bat pollination occurs) on 60.5% of the plants. By visiting the CFNs, the ants indirectly protect the flowers--but not the plant foliage--from herbivorous insects. We show that this protection is provided mostly by species nesting in dried-out stems, predominantly Pseudomyrmex gracilis. That dried-out stems remain standing for years and are regularly replaced results in an opportunistic, but stable association where colonies are sheltered by one generation of dead C. alatus while the live individuals nearby, belonging to the next generation, provide them with nectar; in turn, the ants protect their flowers from herbivores. We suggest that the investment in wood by C. alatus individuals permitting still-standing, dried-out stems to shelter ant colonies constitutes an extended phenotype because foraging workers protect the flowers of live individuals in the same patch. Also, through this process these dried-out stems indirectly favor the reproduction (and so the fitness) of the next generation including both their own offspring and that of their siblings, all adding up to a potential case of inclusive fitness in plants.

Delayed selfing in an alpine biennial Gentianopsis paludosa (Gentianaceae) in the Qinghai-Tibetan plateau.

Delayed selfing could provide ovules with an opportunity to be fertilized as a means of "pollination assurance" before the flowers wilt. It could, thus, be regarded as an adaptation to unpredictable pollinator environments. Within the alpine biennial Gentianopsis paludosa, the showy flowers and herkogamy at the early stage of a flower's life cycle may favor outcrossing. As the flower ages, anthers contact the central stigma due to the elongation of all filaments, resulting in autonomous selfing. Flower visitors are extremely rare in a high altitude population; and examination of the mating system indicates that G. paludosa is self-pollinated under natural conditions in this population. While at the lower altitude, the bumblebee visitation rate is relatively high but possibly unreliable. Stigma receptivity is the highest on the third day of anthesis, and decreases thereafter. Pollen viability is the highest when flowers open, and gradually decreases later. Self-pollination of G. paludosa occurs at the late stage of a flower's lifecycle when stigma receptivity and pollen viability have both decreased, suggesting delayed selfing and assurance of seed production. This delayed selfing could assure seed production under the constraints of pollinator scarcity, but ensure outcrossing when pollinators were available. Such a flexible pollination mechanism is highly adaptive in the alpine environment of the Qinghai-Tibetan Plateau.

Nectar sugar composition and volumes of 47 species of Gentianales from a southern Ecuadorian montane forest.

BACKGROUND AND AIMS: This study investigates 47 taxonomically related species (Gentianales), all native to a tropical montane forest in southern Ecuador, in terms of nectar chemistry and nectar volumes in relation to pollination biology. METHODS: Nectar volumes of covered (24-h production) and uncovered (standing crop) flowers were measured in the natural habitat. Sucrose, fructose and glucose were quantified in the nectar using high performance liquid chromatography. Flower visitors were observed. KEY RESULTS: Nectar sugar concentration did not differ significantly among the pollination syndromes. Regarding sugar composition, the only significant differences were found in chiropterophilous and myiophilous flowers, which had a significantly lower sugar ratio than sphingophilous flowers. A separation of chiropterophilous and myiophilous flowers from the other pollination syndromes is further substantiated by non-linear multidimensional scaling using the chord-normalized expected species shared index of dissimilarity based on nectar sugar compositions. The matrix test revealed no correlation of observed floral visitors to nectar concentrations; however, a weak significant correlation was found between floral visitors and nectar sugar compositions. The nectar volumes of covered and uncovered flowers are related to, and differ significantly among, pollination syndromes. Matrix tests revealed correlation between floral visitors and nectar volume of covered flowers and, to a lesser extent, of uncovered flowers. CONCLUSIONS: Sucrose is the predominant floral nectar sugar in the order Gentianales, suggesting that nectar sugar composition is a conservative characteristic. However, some degree of an adaptive convergence of floral nectar compositions to principal pollinator type within the constraints set by phylogenetic history is likely. The driving force to visitation appears to be the volume of nectar the visitor can expect to consume.