Pollination by passerine birds: why are the nectars so dilute?

Bird-pollinated flowers are known to secrete relatively dilute nectars (with concentrations averaging 20–25% w/w). Many southern African plants that are pollinated by passerine birds produce nectars with little or no sucrose. Moreover, these hexose nectars are extremely dilute (10–15%). This suggests a link between sugar composition and nectar concentration. Nectar originates from sucrose-rich phloem sap, and the proportion of monosaccharides depends on the presence and activity of invertase in the nectary. Hydrolysis of sucrose increases nectar osmolality and the resulting water influx can potentially convert a 30% sucrose nectar into a 20% hexose nectar, with a 1.56 times increase in volume. Hydrolysis may also increase the gradient for sucrose transport and thus the rate of sugar secretion. When sucrose content and refractometer data were compared, some significant correlations were seen, but the occurrence of sucrose-rich or hexose-rich nectars can also be explained on phylogenetic grounds (e.g. Erythrina and Protea). Hexose nectars may be abundant enough to drip from open flowers, but evaporation leads to much variability in nectar concentration and increases the choices available to pollinators.     

Floral nectary structure and nectar composition in Eccremocarpus scaber (Bignoniaceae), a hummingbird-pollinated plant of central Chile

Surface features, anatomy, and ultrastructure of the floral nectary of Eccremocarpus scaber (Bignoniaceae), pollinated predominantly by the largest-known hummingbird (Patagona gigas gigas), were studied together with nectar sugar content and secretion rate. The annular disk nectary comprises epidermis, secretory and ground parenchyma with intercellular spaces, and branched vascular bundles terminating in the secretory parenchyma where only phloem is found. Amyloplasts and vacuoles increase in size throughout development, the latter becoming sites of organelle degradation. Transferlike cells in nectary phloem and P-proteinlike fibrillar material in phloem parenchyma were observed. Flowers produced around 32 μl of nectar (mostly after anthesis) with 11 mg of sugar composed of fructose, glucose, sucrose, and maltose in a ratio of 0.34:0.32:0.17:0.17. Morphological studies as well as the presence of maltose and glucose in nectar suggest storage of the originally phloem-derived sugars as starch with its subsequent hydrolysis. The low sucrose/hexose ratio (0.25) and high nectary secretion force (nectar per flower biomass) observed places E. scaber close to large-bodied bat-pollinated plants. A hypothesis based on nectar origin and nectar secretion is advanced to explain pollinator-correlated variation in sucrose/hexose ratio.     

The nectar of the Strelitzia reginae flower

The nectar of Strelitzia reginae Ait. was analysed using enzymatic methods and found to contain glucose, fructose and sucrose. Sugar composition changed considerably over the nectar producing period: there was an increase in the amount of glucose (41%) and fructose (32%) between the early and middle stage of secretion and thereafter a decrease of 13 and 24%, respectively, towards the end of secretion. Although the amount of sucrose secreted was initially as much as the glucose and fructose combined, it subsequently decreased, first by 14% and then by 70% at the end of the secretory period so that, whereas in the initial stages of secretion sucrose was quantitatively the dominant sugar, glucose and fructose made up the major part of the nectar as secretion reached its conclusion.
The amounts of potassium and sodium remained at the same low level (around 150 and 30 μg g^-1 [w/v) respectively) throughout secretion, while calcium (initially 18 μg g^-1) and magnesium (initially 8.0 μg g^-1) increased by 47 and 56% respectively, between the early and late stages of secretion. No free amino acids, inorganic phosphate or iron could be detected. Enzymatic analysis revealed only a trace amount of starch. Transmission electron micrographs from both immature and mature plants, however, showed starch grains among other cytoplasmic remnants in the nectary lumen. Mitochondria, vesicles, lipid droplets and ribosomes could also be identified among the luminal cytoplasmic remnants.     

Characteristics of Nectar Secretion by the Extrafloral Nectaries of Ricinus communis

The characteristics of nectar secretion by excised extrafloralnectaries of Ricinus have been examined. Secreted nectar wasfound to contain three sugars: sucrose, glucose and fructose,with glucose and fructose occurring in a 1: 1 ratio. All threesugars supported secretion when used in the culture medium andthe yield of nectar sugar was found to be concentration-dependent.Other sugar sources failed to support secretion. Experimentsusing ¹⁴C-sugars and ¹⁴CO₂ fed to intact plants allowed themovement of sugars through the nectary to be examined. Sucrosesynthesis occurs when excised glands are fed glucose and thisoccurs very early in the transport through the nectary. Themain sugar transported was sucrose, with little hydrolysis occurringuntil the final step of secretion. There was no evidence thatsucrose hydrolysis occurs either by invertase in the nectaror by a microbial flora. Inhibitors of respiration were foundto inhibit secretion as did anaerobiosis. Temperature also hada marked effect, with a temperature coefficient of 1.8. However,secretion of sucrose was not affected by anaerobic conditions,low temperatures or inhibitors of respiration as markedly asthat of glucose and fructose. Electron microscopy revealed the presence of a thickened andheavily stained wall at the inner border of the secretory epidermallayer. This wall contained numerous plasmodesmata at a frequencyof 14 per µm² and may represent an apoplastic barrier.Light microscope cytochemistry revealed that acid phosphataseis primarily located in the nectiferous tissue, while ATPaseis concentrated in the epidermis. The possibility that the nectarycontains two pathways for sucrose secretion, both apoplasticand symplastic, is discussed. Key words: Invertase, nectary, plasmodesmata, Ricinus communis, sucrose     

Intra-plant variation in nectar sugar composition in two Aquilegia species (Ranunculaceae): contrasting patterns under field and glasshouse conditions

BACKGROUND AND AIMS: Intra-specific variation in nectar chemistry under natural conditions has been only rarely explored, yet it is an essential aspect of our understanding of how pollinator-mediated selection might act on nectar traits. This paper examines intra-specific variation in nectar sugar composition in field and glasshouse plants of the bumblebee-pollinated perennial herbs Aquilegia vulgaris subsp. vulgaris and Aquilegia pyrenaica subsp. cazorlensis (Ranunculaceae). The aims of the study are to assess the generality of extreme intra-plant variation in nectar sugar composition recently reported for other species in the field, and gaining insight on the possible mechanisms involved. METHODS: The proportions of glucose, fructose and sucrose in single-nectary nectar samples collected from field and glasshouse plants were determined using high performance liquid chromatography. A hierarchical variance partition was used to dissect total variance into components due to variation among plants, flowers within plants, and nectaries within flowers. KEY RESULTS: Nectar of the two species was mostly sucrose-dominated, but composition varied widely in the field, ranging from sucrose-only to fructose-dominated. Most intra-specific variance was due to differences among nectaries of the same flower, and flowers of the same plant. The high intra-plant variation in sugar composition exhibited by field plants vanished in the glasshouse, where nectar composition emerged as a remarkably constant feature across plants, flowers and nectaries. CONCLUSIONS: In addition to corroborating the results of previous studies documenting extreme intra-plant variation in nectar sugar composition in the field, this study suggests that such variation may ultimately be caused by biotic factors operating on the nectar in the field but not in the glasshouse. Pollinator visitation and pollinator-borne yeasts are suggested as likely causal agents.     

Pollinator adaptation and the evolution of floral nectar sugar composition

A long‐standing debate concerns whether nectar sugar composition evolves as an adaptation to pollinator dietary requirements or whether it is ‘phylogenetically constrained’. Here, we use a modelling approach to evaluate the hypothesis that nectar sucrose proportion (NSP) is an adaptation to pollinators. We analyse ~ 2100 species of asterids, spanning several plant families and pollinator groups (PGs), and show that the hypothesis of adaptation cannot be rejected: NSP evolves towards two optimal values, high NSP for specialist‐pollinated and low NSP for generalist‐pollinated plants. However, the inferred adaptive process is weak, suggesting that adaptation to PG only provides a partial explanation for how nectar evolves. Additional factors are therefore needed to fully explain nectar evolution, and we suggest that future studies might incorporate floral shape and size and the abiotic environment into the analytical framework. Further, we show that NSP and PG evolution are correlated – in a manner dictated by pollinator behaviour. This contrasts with the view that a plant necessarily has to adapt its nectar composition to ensure pollination but rather suggests that pollinators adapt their foraging behaviour or dietary requirements to the nectar sugar composition presented by the plants. Finally, we document unexpectedly sucrose‐poor nectar in some specialized nectarivorous bird‐pollinated plants from the Old World, which might represent an overlooked form of pollinator deception. Thus, our broad study provides several new insights into how nectar evolves and we conclude by discussing why maintaining the conceptual dichotomy between adaptation and constraint might be unhelpful for advancing this field.     

Coloured and Toxic Nectar: Feeding Choices of the Madagascar Giant Day Gecko,Phelsuma grandis

Coloured nectar is a rare phenomenon best known from islands and insular habitats. Islands are also known for lizard pollination, where coloured nectar potentially acts as a visual cue to attract pollinators, advertising the sweet reward. However, nectar may also contain secondary metabolites with toxic or deterrent effects. The aim of this study was to determine which factors are important as artificial nectar choice determinants to the Madagascar giant day gecko, Phelsuma grandis, an island pollinator: artificial nectar colour, artificial nectar colour saturation, artificial nectar conspicuousness and/or the presence of the alkaloid nicotine. Coloured artificial nectar and the darkest artificial nectar colour saturation were found to be important visual cues for the geckos, while the contrast between artificial nectar and petal colour was not. Geckos were deterred only by high nicotine concentrations (1000 μm in 0.63 m sucrose) and may even prefer low nicotine concentrations to sucrose‐only solutions. Given their overall fondness for sugar solutions, Madagascar giant day geckos are likely to be important pollinators of Malagasy plant species that produce enough nectar to attract them, and plants with coloured nectar and/or secondary metabolites may have evolved those traits to attract the geckos in particular.     

Variation in nectar volume and composition of Impatiens capensis at the individual,plant, and population levels

Although the volume and chemical composition of nectars are known to vary among plant species and to affect pollinator response to plants, relatively little is known of the variation in volume, and sugar and amino acid composition within species. We collected nectar from Impatiens capensis in a nested design: three flowers from each of three plants from each of three populations. This design enabled us to quantify variation within individual plants, among plants within populations, and among populations. Using high performance liquid chromatography, we analyzed the sugar and amino composition of the 27 flowers. Analysis of variance showed that none of the parameters (volume, concentrations of three sugars and 24 amino compounds) varied within individuals. Variation in nectar volume was not significant among plants but was nearly significant among populations. Of the three sugars detected (sucrose, glucose, and fructose), the only significant variation was that of sucrose among populations. Concentrations of 12 amino compounds varied significantly at the plant level while 7 amino compounds varied among populations. The results indicate that: (1) pooling of nectar samples from flowers of individual plants can be an acceptable methodology for those seeking to understand within species variation; (2) amino compounds appear to vary more than either volumes or sugar concentrations; (3) future studies should assess how much of the observed variation is due to genetic versus environmental differences; (4) additional studies should examine the geographic variation in nectar parameters and pollinators of I. capensis in order to assess the role different pollinators play in shaping nectar composition.     

The Role of Jasmonates in Floral Nectar Secretion

Plants produce nectar in their flowers as a reward for their pollinators and most of our crops depend on insect pollination, but little is known on the physiological control of nectar secretion. Jasmonates are well-known for their effects on senescence, the development and opening of flowers and on plant defences such as extrafloral nectar. Their role in floral nectar secretion has, however, not been explored so far. We investigated whether jasmonates have an influence on floral nectar secretion in oil-seed rape, Brassica napus. The floral tissues of this plant produced jasmonic acid (JA) endogenously, and JA concentrations peaked shortly before nectar secretion was highest. Exogenous application of JA to flowers induced nectar secretion, which was suppressed by treatment with phenidone, an inhibitor of JA synthesis. This effect could be reversed by additional application of JA. Jasmonoyl-isoleucine and its structural mimic coronalon also increased nectar secretion. Herbivory or addition of JA to the leaves did not have an effect on floral nectar secretion, demonstrating a functional separation of systemic defence signalling from reproductive nectar secretion. Jasmonates, which have been intensively studied in the context of herbivore defences and flower development, have a profound effect on floral nectar secretion and, thus, pollination efficiency in B. napus. Our results link floral nectar secretion to jasmonate signalling and thereby integrate the floral nectar secretion into the complex network of oxylipid-mediated developmental processes of plants.     

The modern science of ambrosiology: in honour of Herbert and Irene Baker

 Herbert and Irene Baker were a team. Their contributions to pollination biology have been many, but it is in the area of nectar its chemical composition, functional and ecological significance, and value in taxonomy and evolution that we honour them here. Although nectar can be described as simply a sugary secretion from plants, we know that it is more than that. Herbert and Irene explored the depths of nectar chemistry, analysing sugar components, non-sugar constituents that can benefit nectarivorous animals (amino acids, lipids, antioxidants, proteins, minerals, vitamins) or that may be deleterious (alkaloids, phenolics, other toxins, salts, and perhaps some other amino acids). They related the ratios of disaccharide (sucrose) to monosaccharides (glucose and fructose), and the amounts of amino acids, and other constituents in nectar to pollinator types and theorized on the reasons for the trends they observed. They noted tight relationships between nectar constituents and plant systematics, and additive effects of some constituents after hybridization. In all their nectar studies made over a quarter century, evolution and ecology were forefront. This scholarly couple made an indelible mark. Received November 12, 2002; accepted January 17, 2003 Published online: June 2, 2003     

A model for nectar secretion in animal-pollinated plants

Summary A game theoretic model was developed for nectar secretion in animal-pollinated plants in order to examine how the total amount of resources allocated to flowers affects the spread of nectarless plants. It was assumed that pollinators concentrate on patches whose nectar rewards are relatively large compared to other patches and if pollinators visit a patch, they concentrate on the plants whose nectar rewards are relatively large compared to other plants in the patch. It was shown that plants are more likely to secrete nectar in populations where the total amount of resources allocated to flowers is large. It was also shown that strong interplant competition, strong interpatch competition and the nectar discrimination of the pollinators are also important factors for nectar secretion. However, if the total amount of resources allocated to flowers is sufficiently large, plants would secrete nectar even if competition is not very strong and nectar discrimination is not so precise.     

Why do ants shift their foraging from extrafloral nectar to aphid honeydew?

When aphids parasitize plants with extrafloral nectaries (EFNs) and aphid colony size is small, ants frequently use EFNs but hardly tend aphids. However, as the aphid colony size increases, ants stop using EFNs and strengthen their associations with aphids. Although the shift in ant behavior is important for determining the dynamics of the ant–plant–aphid interaction, it is not known why this shift occurs. Here, we test two hypotheses to explain the mechanism responsible for this behavioral shift: (1) Extrafloral nectar secretion changes in response to aphid herbivory, or (2) plants do not change extrafloral nectar secretion, but the total reward to ants from aphids will exceed that from EFNs above a certain aphid colony size. To judge which mechanism is plausible, we investigated secretion patterns of extrafloral nectar produced by plants with and without aphids, compared the amount of sugar supplied by EFNs and aphids, and examined whether extrafloral nectar or honeydew was more attractive to ants. Our results show that there was no inducible extrafloral secretion in response to aphid herbivory, but the sugar concentration in extrafloral nectar was higher than in honeydew, and more ant workers were attracted to an artificial extrafloral nectar solution than to an artificial aphid honeydew solution. These results indicate that extrafloral nectar is a more attractive reward than aphid honeydew per unit volume. However, even an aphid colony containing only two individuals can supply a greater reward to ants than EFNs. This suggests that the ant behavioral shift may be explained by the second hypothesis.     

VARIATION IN NECTAR SUGAR COMPOSITION AT THE INTRAPLANT LEVEL IN IPOMOPSIS LONGIFLORA (POLEMONIACEAE)

The nectar sugar composition of individual flowers from single plants in both greenhouse and field populations was determined by High-Performance Liquid Chromatography (HPLC). Intraplant variability in nectar sugar composition of greenhouse-grown plants was significantly greater than can be attributed to the analytical methodology. Individual greenhouse-grown plants had significantly different sugar compositions, suggesting genetic differences. Under field conditions the situation was complex. Variation among flowers from single plants in field populations was significantly greater than that of plants under greenhouse conditions. Unprotected flowers of some field populations were more variable at the intraplant level than others. Field experiments showed that, in untreated (open-visited) flowers, the percent sucrose relative to other sugars declined significantly and coefficients of variability increased significantly with flower age. However, nectars never exposed to the natural environment did not change significantly with flower age. Further, within single plants it was found that percent sucrose in nectars varied in a small but significant way during the growing season. The data suggest that, in this species, both intrinsic and extrinsic factors affect nectar sugar composition and it is desirable to obtain samples from numerous flowers to accurately characterize the nectar sugar composition of an individual under field conditions.     

Extreme intraplant variation in nectar sugar composition in an insect-pollinated perennial herb

Variation in nectar chemistry among plants, flowers, or individual nectaries of a given species has been only rarely explored, yet it is an essential aspect to our understanding of how pollinator-mediated selection might act on nectar traits. This paper describes variation in nectar sugar composition in a population of the perennial herb Helleborus foetidus (Ranunculaceae) and dissects it into components due to variation among plants, flowers of the same plant, and nectaries of the same flower. The proportions of sucrose, glucose, and fructose in single-nectary nectar samples collected at two times in the flowering season were determined using high performance liquid chromatography (HPLC). Sugar composition varied extensively among nectaries, and nearly all combinations of individual sugars were recorded. Population-wide variance was mainly accounted for by variation among flowers of the same plant (56% of total), nectaries of the same flower (30%), and only minimally by differences among plants (14%). In absolute terms, intraplant variation was similar to or greater than that ordinarily reported in interspecific comparisons. Results suggest that the prevailing notion of intraspecific constancy in nectar sugar composition may be unwarranted for some species and that more elaborate nectar sampling designs are required to detect and appropriately account for extensive within-plant variance. Within-plant variation in nectar sugar composition will limit the ability of pollinators to exert selection on nectar chemistry in H. foetidus and may be advantageous to plants by reducing the number of flowers visited per foraging bout by variance-sensitive, risk-averse pollinators.     

Nectar production and carbohydrate composition across floral sexual phases: contrasting patterns in two protandrous Aconitum species (Delphinieae,Ranunculaceae)

In dichogamous species, it is necessary that pollinators are attracted both to male- and female-phase flowers in order to achieve effective cross-pollination. We investigated, over a two-year period, how nectar production and composition differs in protandrous flowers of Aconitum lycoctonum L. and Aconitum carmichaelii Debeaux, two species originating from different geographical regions. Flowers of A. carmichaelii secreted approx. 2.5-fold more nectar than flowers of A. lycoctonum. The nectar of A. carmichaelii was sucrose-dominant (87.6:9.5:2.9, sucrose:fructose:glucose), whereas sucrose-rich nectar, lacking glucose, occurred in A. lycoctonum (39.9:60.1, sucrose:fructose). Total sugar concentration was similar (53%, on average) in both species. These species also showed contrasting patterns in nectar production and composition between the floral sexual phases. On average, in A. carmichaelii, male-phase flowers produced 2.4-fold more nectar than female-phase flowers in the two years of study, and nectar was sucrose-dominant, irrespective of sexual phase. By contrast, nectar production in A. lycoctonum was biased towards the male phase in one year of study and towards the female phase in the other, and whereas nectar was sucrose-dominant during the male-phase, it was fructose-rich during the female phase, indicating sucrose re-absorption. Although the characteristics of nectar in these Aconitum species indicate pollination by bumblebees, it is possible that their biogeographical history, and not pollinator selection alone, is important in understanding the lack of glucose in A. lycoctonum nectar. Variability in nectar production and/or carbohydrate composition between sexual phases suggests indirectly that nectar traits complement each of the latter. Nevertheless, further research is required if we are to understand the significance of these disparities in pollination ecology, i.e. for promoting pollinator movements between flowers and plants in order to achieve cross-pollination and avoid inbreeding.     

Nectary Structure, Nectar Secretion Patterns and Nectar Composition in Two Helleborus Species

Abstract: The morphological and cytological characteristics of nectaries of Helleborus foetidus and H. bocconei during the secretory period are reported. The nectaries are derived from modified petals and secrete nectar continuously for about 20 days; they consist of a single layered epidermis, nectar-producing parenchyma and photosynthesizing parenchyma. Nectar secretion is holocrine and the nectar is released by rupture of the wall and cuticle of each epidermal cell. The nectaries of the two species differ in number and external morphology. In H. foetidus, secretion begins before anthesis and secretion rate decreases with nectary age. In H. bocconei it begins on the day of anthesis and proceeds at a constant rate. The nectar has a high sugar content, mainly sucrose, and also contains lipids and proteins.     

The extrafloral nectaries of cowpea (Vigna unguiculata (L.) Walp.) II. Nectar composition,origin of nectar solutes,and nectary functioning

Nectar was collected from the extrafloral nectaries of leaf stipels and inflorescence stalks, and phloem sap from cryopunctured fruits of cowpea plants. Daily sugar losses as nectar were equivalent to only 0.1–2% of the plant's current net photosynthate, and were maximal in the fourth week after anthesis. Sucrose:glucose:fructose weight ratios of nectar varied from 1.5:1:1 to 0.5:1:1, whereas over 95% of phloem-sap sugar was sucrose. [¹⁴C]Sucrose fed to leaves was translocated as such to nectaries, where it was partly inverted to [¹⁴C]glucose and [¹⁴C]fructose prior to or during nectar secretion. Invertase (EC 3.2.1.26) activity was demonstrated for inflorescence-stalk nectar but not stipel nectar. The nectar invertase was largely associated with secretory cells that are extruded into the nectar during nectary functioning, and was active only after osmotic disruption of these cells upon dilution of the nectar. The nectar invertase functioned optimally (phloem-sap sucrose as substrate) at pH 5.5, with a starting sucrose concentration of 15% (w/v). Stipel nectar was much lower in amino compounds relative to sugars (0.08–0.17 mg g^-1 total sugar) than inflorescence nectar (22–30 mg g^-1) or phloem sap (81–162 mg g^-1). The two classes of nectar and phloem sap also differed noticeably in their complements of organic acids. Xylem feeding to leaves of a range of ¹⁴C-labelled nitrogenous solutes resulted in these substrates and their metabolic products appearing in fruit-phloem sap and adjacent inflorescence-stalk nectar. ¹⁴C-labelled asparagine, valine and histidine transferred freely into phloem and appeared still largely as such in nectar. ¹⁴C-labelled glycine, serine, arginine and aspartic acid showed limited direct access to phloem and nectar, although labelled metabolic products were transferred and secreted. The ureide allantoin was present in phloem, but absent from both types of nectar. Models of nectary functioning are proposed.     

Temporal changes in floral nectar production, reabsorption, and composition associated with dichogamy in annual caraway (Carum carvi; Apiaceae)

The dynamics of nectar production were studied in perfect florets of two varieties (Karzo, Moran) of annual caraway (Carum carvi L., Apiaceae). Florets were protandrous and strongly dichogamous, lasting 7-15 d but producing nectar from the stylopodia for 4-12 d, in an interrupted fashion. Nectar secretion began during a floret's phase of stamen elongation and anther dehiscence. After reabsorption of uncollected nectar, at which point nectary surfaces were completely dry, the two styles elongated and a second bout of secretion commenced during the female phase, up to 5 d later, when a floret became receptive to pollination. During the male and female phases, respectively, 0.392 ± 0.064 μL and 1.083 ± 0.261 μL of nectar of similar solute concentration (844 mg/mL) was produced per ten florets. On a daily basis, florets yielded 1.5-fold more nectar in the female than during the male phase. First-time nectar removal throughout the female phase did not match the sum of nectar quantities from male and female phases combined, suggesting that under natural conditions, any uncollected male-phase nectar, once reabsorbed, is not made available to visitors of the same florets when in the female phase. Nectar-sugar composition differed between bouts of secretion; it was hexose-rich (59.6% fructose, 26.9% glucose, 13.6% sucrose) initially, but hexose-dominant (70.2, 26.8, 3.1) during the female phase. A 5.7-fold difference in mean nectar production per floret occurred among plants.     

A cell wall invertase controls nectar volume and sugar composition

Nectar volume and sugar composition are key determinants of the strength of plant–pollinator mutualisms. The main nectar sugars are sucrose, glucose and fructose, which can vary widely in ratio and concentration across species. Brassica spp. produce a hexose-dominant nectar (high in the monosaccharides glucose and fructose) with very low levels of the disaccharide sucrose. Cell wall invertases (CWINVs) catalyze the irreversible hydrolysis of sucrose into glucose and fructose in the apoplast. We found that BrCWINV4A is highly expressed in the nectaries of Brassica rapa. Moreover, a brcwinv4a null mutant: (i) has greatly reduced CWINV activity in the nectaries; (ii) produces a sucrose-rich nectar; but (iii) with significantly less volume. These results definitively demonstrate that CWINV activity is not only essential for the production of a hexose-rich nectar, but also support a hypothetical model of nectar secretion in which its hydrolase activity is required for maintaining a high intracellular-to-extracellular sucrose ratio that facilitates the continuous export of sucrose into the nectary apoplast. The extracellular hydrolysis of each sucrose into two hexoses by BrCWINV4A also likely creates the osmotic potential required for nectar droplet formation. These results cumulatively indicate that modulation of CWINV activity can at least partially account for naturally occurring differences in nectar volume and sugar composition. Finally, honeybees prefer nectars with some sucrose, but wild-type B. rapa flowers were much more heavily visited than flowers of brcwinv4a, suggesting that the potentially attractive sucrose-rich nectar of brcwinv4a could not compensate for its low volume.