Sunflower pollen reduces a gut pathogen in worker and queen but not male bumble bees

1. Social insect castes and sexes differ in many ways, including morphology, behavior, and sometimes ploidy level. Recent studies have found that consuming sunflower pollen reduces the gut pathogen Crithidia bombi in workers of the common eastern bumble bee (Bombus impatiens). Here, this work is extended to the reproductive individuals that represent colony fitness – males and queens – to assess if the medicinal effects of sunflower pollen vary with bee caste and sex. 2. This study examined the effect of sunflower pollen compared to a diverse wildflower pollen mix on infection in worker, male, and daughter queen commercial B. impatiens. Bees were infected, fed either sunflower pollen or wildflower pollen for 7 days, and then infection levels were assessed. 3. Compared to wildflower pollen, sunflower pollen dramatically reduced Crithidia infection in workers and daughter queens, but not males. Infection levels were very low for both diets in males; this could be due to low pollen consumption or other mechanisms. 4. Reducing Crithidia infection in young queens before they undergo hibernation is important for population dynamics since infected queens are less likely to survive hibernation, and those that do are less likely to successfully establish a nest the following spring. Because sunflowers bloom in late summer when new queens are emerging, sunflowers could provide an important dietary component for queens during this critical life stage. Deepening our understanding of how diet impacts pathogens in reproductive bees, as well as workers, is crucial to maintain healthy pollinator populations.     

The life-history impact and implications of multiple parasites for bumble bee queens

Most studies of the consequences of parasitism on fitness have examined single host–parasite systems. However, parasitological studies show that most hosts are constantly challenged by a complex parasite community. Thus, neither the response of hosts to individual parasite species nor the individual impact of these parasite species is likely to be as unconstrained as studies of single host–parasite systems might suggest. In this study, the parasite community structure in spring queens of the common European bumble bee, Bombus pratorum, was assessed. By capturing queens and allowing them to rear colonies in the laboratory, the relative impact of different parasite species on fitness across the annual host life-cycle could be examined. Of 160 queens, 67% were parasitised by one or more members of a five-species parasite community. The impact of parasites varied from being highly virulent to undetectable under benign laboratory conditions. The majority of multi-parasite infections involved a high impact parasite, which resulted in the removal of associated parasites from the host population. This study shows that, whilst multiple infections occur within individual hosts, most parasites act individually on their hosts. However, multiple parasite species in the host population may constrain the host population’s ability to adapt to single parasite species.     

Orange, yellow and white-cream: inheritance of carotenoid-based colour in sunflower pollen

Inheritance of pollen colour was studied in sunflower (Helianthus annuus L.) using three distinct pollen colour morphs: orange, yellow and white‐cream. Orange is the most common colour of sunflower pollen, while the yellow morph is less frequent. These two types were observed in the inbred lines F11 and EF2L, respectively. White‐cream pollen is a rare phenotype in nature, and was identified in a mutant, named white‐cream pollen, recovered in the R₂ generation of an in vitro regenerated plant. The F11 inbred line was used as starting material for in vitro regeneration. The carotenoid content of these three pollen morphs differed, and was extremely reduced in white‐cream pollen. The phenotype of F₁ populations obtained by reciprocal crosses revealed that the orange trait was dominant over both white‐cream and yellow. Segregation of F₂ populations of both crosses, orange × yellow and orange × white‐cream, approached a 3:1 ratio, indicating the possibility of simple genetic control. By contrast, a complementation cross between the two lines with white‐cream and yellow pollen produced F₁ plants with orange pollen. The F₂ populations of this cross‐segregated as nine orange: four white‐cream: four yellow. A model conforming to the involvement of two unlinked genes, here designated Y and O, can explain these results. Accessions with yellow pollen would have the genotype YYoo, the white‐cream pollen mutant would have yyOO and the accession with orange pollen would have YYOO. Within F₂ populations of the cross white‐cream × yellow a new genotype, yyoo, with white‐cream pollen was scored. The results of the cross yyoo × YYoo produced only F₁ plants with yellow pollen, supporting a recessive epistatic model of inheritance between two loci. In this model, yy is epistatic on O and o. In F₂ populations, the distributions of phenotypic classes suggested that the genetic control of carotenoid content is governed by major genes, with large effects segregating in a background of polygenic variation. These three pollen morphs can provide insight into the sequence in which genes act, as well into the biochemical pathway controlling carotenoid biosynthesis in anthers and the transfer of these different pigments into pollenkitt.     

Contrasting movement patterns of nectar-collecting and pollen-collecting bumble bees (Bombus terricola) on fireweed (Chamaenerion angustifolium) inflorescences

Abstract. 1. Movements of nectar and pollen-foraging bumble bees on inflorescences of Chamaenerion angustifolium (L.) J. Holub (fireweed or rosebay willow herb) were compared with predictions based on reward distributions and optimality principles. Observations suggest that nectar and pollen-gathering bumble bees behave according to the same set of reward maximization criteria when foraging from flowers of this species.
2. Both kinds of foragers matched their arrival points with the vertical positions on inflorescences in which the densities of their respective food resources were greatest. For nectar-foragers, this point was located at the lowest tier of flowers, whereas for pollen-foragers it was found in the middle of the inflorescences. Nectar and pollen-foraging bees both moved upward on inflorescences following gradients from high to low reward availability.
3. Nectar-foragers responded to decreases in inflorescence size over the season by reducing the number of flower visits made on each raceme. Number of flowers visited by pollen-foragers was low throughout and reflected the scarcity of male-phase flowers on racemes. Flower revisitation rates were low for both kinds of workers, but were slightly higher for those collecting pollen.     

Effects of nitrate on influx, efflux and translocation of potassium in young sunflower plants

Influx, efflux and translocation of K⁺(⁸⁶Rb) were studied in the roots of sunflower seedlings ( Helianthus annuus L. cv. Uniflorus) treated with 0–4.0 m M NO₃⁻ during a 9 day growth period or a 24 h pretreatment period. Roots treated with high levels of NO₃⁻ absorbed and translocated more K⁺(⁸⁶Rb) than seedlings treated with low levels of NO₃⁻. The content of K⁺ in the shoots was, however, higher in seedlings treated with low levels of NO₃⁻, indicating a low rate of retranslocation of K⁺ in those plants. K⁺(⁸⁶Rb) efflux was highest into the low-NO₃⁻ solutions. All effects on K⁺(⁸⁶Rb)-fluxes were more obvious in high-K plants than in low-K plants. The results are discussed in relation to the Dijkshoorn-Ben Zioni hypothesis for K⁺+ NO₃⁻-uptake and translocation in plants.     

Influence of pollen selection for Alternaria helianthi resistance on the progeny performance against leaf blight in sunflower (Helianthus annuus L.)

Seven genotypes of sunflower, including populations and hybrids, showing differential susceptibility to Alternaria leaf and stem blight were crossed to CMS 234A with pollen selection and without pollen selection. The pathogen culture filtrate was used as selective pressure at stylar tissue by applying it 1 h before pollination. Distilled water applied to stigmas and styles served as control. Two sets of seven hybrids, one set from selective and the other from non-selective fertilization, were evaluated for reaction to Alternaria leaf and stem blight during the rainy season under natural epiphytotic conditions. Selection for resistant pollen on the stigmatic surface resulted in a corresponding increase in progeny resistance. The study demonstrates the transmission of the selected trait from the pollen generation to progeny. Further, it was observed that the effect of pollen selection was high in the progenies of moderately resistant parents compared to progenies of highly susceptible parents. The effect of successive pollen selection was studied by backcrossing the progeny derived through selective fertilization to the fertile parent using selective fertilization. Successive pollen selection further improved disease resistance of progeny. However, the improvement was not very great. Hence, repeated cycles of selection are required to achieve a useful level of resistance in the case of sunflower, since resistance is polygenetically controlled. Received: 17 May 1999 / Revision accepted: 13 September 1999     

Sex, horizontal transmission, and multiple hosts prevent local adaptation of Crithidia bombi, a parasite of bumblebees (Bombus spp.)

Local adaptation within host-parasite systems can evolve by several non-exclusive drivers (e.g., host species-genetic adaptation; ecological conditions-ecological adaptation, and time-temporal adaptation). Social insects, especially bumblebees, with an annual colony life history not only provide an ideal system to test parasite transmission within and between different host colonies, but also parasite adaptation to specific host species and environments. Here, we study local adaptation in a multiple-host parasite characterized by high levels of horizontal transmission. Crithidia bombi occurs as a gut parasite in several bumblebee species. Parasites were sampled from five different host species in two subsequent years. Population genetic tools were used to test for the several types of adaptation. Although we found no evidence for local adaptation of the parasite toward host species, there was a slight temporal differentiation of the parasite populations, which might have resulted from severe bottlenecks during queen hibernation. Parasite populations were in Hardy-Weinberg equilibrium and showed no signs of linkage disequilibrium suggesting that sexual reproduction is an alternative strategy in this otherwise clonal parasite. Moreover, high levels of multiple infections were found, which might facilitate sexual genetic exchange. The detection of identical clones in different host species suggested that horizontal transmission occurs between host species and underpins the lack of host-specific adaptation.     

A quantitative in vitro cultivation technique to determine cell number and growth rates in strains of Crithidia bombi (Trypanosomatidae), a parasite of bumblebees

The protozoan parasite Crithidia bombi and its host, the bumblebee Bombus terrestris, are used as a model system for the study of the evolutionary ecology of host-parasite interactions. In order to study these interactions we established a method for in vitro cultivation of single parasite strains. Additionally, a high-throughput method is developed for the determination of cell numbers in cultures by means of optical density (OD) measurements. The protocol for in vitro cultivation allowed for growing different strains on agar plates as well as in culture medium. A calibration curve for the relationship between cell number and OD has been developed. Subsequently, growth rates for different genotypes of C. bombi have been recorded. Significant differences in the growth rates and generation times between these genotypes were demonstrated. As this might be related to the virulence of the parasite, this relationship may be confirmed by in vivo growth rate determination. In comparison with conventional cell counting, the application of OD measurements allows for high-throughput experiments as the time taken to record each sample is reduced by a factor of 30. The in vitro cultivation method allows for controlled infection experiments in order to study host-parasite interactions.     

Effect of steady torque twisting on the orientation of cortical microtubules in the epidermis of the sunflower hypocotyl

Orientation of cortical microtubules (cMTs) is suggested to be affected by mechanical stress existing in cell walls. However, in mutants exhibiting helical (chiral) growth, there is a correlation between orientation of cMTs in outer tissues and helical growth direction. The aim of this research was to examine the effect of a chiral mechanical stimulation on cMTs. For this purpose, the orientation of cMTs was investigated in hypocotyls subjected to either a right- or a left-handed twist, resulting from a steady torque. cMTs were visualised in fixed material using the immunofluorescence method. The cMTs in untouched control hypocotyls were mostly transverse with respect to the cell long axis. In immobilised, but not twisted control hypocotyls, the transverse orientation was also most frequent, while applied twisting resulted in a change in cMT orientation from transverse to oblique. The data provide additional evidence that changes in tissue stress can be reorganized by cortical microtubules.     

Distribution of a model biocontrol agent (Serenade® MAX) in apple and pear by mason bees and bumble bees

1. Biocontrol agents (BCAs) are commonly sprayed on flowering pipfruit trees to prevent them from getting infected by various pathogens. By entomovectoring, BCAs can be directly delivered onto the flowers. However, we currently lack knowledge on the distribution dynamics of BCAs by pollinators.
2. Here, managed bees, both bumble bees (Bombus terrestris) and mason bees (Osmia bicornis and Osmia cornuta), were placed in the vicinity of flowering pipfruit trees (pear -‘Conference’, and apple—‘Svatava’ and ‘Jonagold’), and this allowed us to investigate the distribution of a model BCA, namely, Serenade® MAX, from spray-inoculated flowers of a centralized tree to non-inoculated flowers of surrounding receiver trees by bees in an experimental setup in outdoor conditions.
3. One hour after inoculation, we detected an enrichment of BCA in the flowers of the receiver trees and this for each tested pipfruit.
4. The distribution of BCA from treated to untreated flowers was homogenous between the receiver trees for ‘Svatava’, while significantly different loads were detected for both ‘Conference’ and ‘Jonagold’, which might be due to differences in environmental factors, and/or bee characteristics.
5. More research is needed to understand the distribution dynamics of BCAs by pollinators in field conditions, such as in commercial orchards or crop fields, and how this could result in an efficient control.

     

Effect of abscisic acid on exudation of sunflower roots as affected by nutrient status, glucose level and aeration

Three-week-old sunflower plants ( Helianthus annuus L. cv. Halcón) grown in nutrient solution at two K⁺ levels (0.3 and 2.5 m M ) were used to study the effect of 4 μ M abscisic acid (ABA) on the transport of K⁺ (Rb⁺) and water to the exuding stream of decapitated plants. Other conditions of the bathing medium of the roots were also assayed, such as presence of 10 m M glucose, aeration and time of ABA application. In the first 2 or 3 h after ABA application, ABA always promoted water and ion fluxes, even under the most unfavorable conditions such as low K⁺ roots without glucose or under anaerobiosis. The ABA-promoting effect on ion and water flow was higher with glucose in the medium. Under anaerobiosis the ABA effect disappeared after 3 h. With glucose and aeration the ABA-promoting effect appeared early and continued for several hours, although the effect decreased with time. If ABA was applied 24 h before excision, the effect was small or even negative. We suggest that ABA acts directly on membranes of certain root cells (endodermal or both endodermal and cortical cells) by increasing their permeability and thus releasing ions. This will decrease cell turgor pressure and, indirectly, the hydraulic conductivity of the whole root. Under conditions of higher hydraulic conductivity, the presence of ions and glucose in the root stimulates the transport of ions into the xylem. and thus increases the osmotic water flow.     

The effects of enhanced levels of calcium on the gravireaction of sunflower hypocotyls

A current hypothesis states that there is a redistribution of wall calcium from the lower to the upper sides of horizontal shoots during gravireaction, and because calcium stiffens walls, the unequal calcium distribution results in differential wall extensibility on the upper and lower sides, and thus, causes unequal growth. If this hypothesis is valid, then saturating the cell walls with calcium should minimize the effect of calcium redistribution, and thereby inhibit gravicurvature and stiffen the walls. To test this hypothesis, sunflower seedlings were grown on agar containing 0 to 50molm^-3 CaCl₂. The wall-bound calcium content of the tissues increased as the external concentration of CaCl₂ increased, and the epidermal layers were saturated with calcium by the 10molm^-3 CaCl₂ treatment. Contrary to the predictions from the hypothesis, the vertical growth and the gravicurvature rate of plants grown in 10molm^-3 CaCl₂ were actually accelerated, and wall extensibility, as measured by the Instron technique, was unaffected. These results contradict the hypothesis, and provide further evidence that wall-bound calcium is not involved in the reaction phase of gravicurvature.     

Food-environment mediates the outcome of specific interactions between a bumblebee and its trypanosome parasite

Specific host-parasite interactions, where the outcome of exposure to a parasite depends upon the genotypic identity of both parties, have implications for understanding host-parasite coevolution and patterns of genetic diversity. Thus, grasping the extent to which these interactions are mediated by environmental changes in a spatially and temporally heterogeneous world is vital. In this study, it is shown that the environment can influence specific host-parasite interactions in the well-studied system of the bumblebee Bombus terrestris and its trypanosome parasite Crithidia bombi. Naturally relevant variation in the quality of the food environment formed a three-way interaction with both host and parasite identity in determining the outcome of infection, with regard to the resistance of the host and the transmission of the parasite. The demonstration of such a host-genotype by parasite-genotype by environment interaction (G(H) x G(P) x E) shows the importance of considering environmental variation when investigating host-parasite interactions. Moreover, such interactions may to some extent explain levels of genetic diversity in natural host-parasite systems owing to the fact that they will create selection mosaics when environments are heterogeneous.     

Interaction between sunflower plants and five isolates of Plasmopara halstedii on the level of pathogenicity

The interaction between sunflower plants showing a high level of quantitative resistance and five Plasmopara halstedii (the causal agent of downy mildew) isolates of several races were studied using five single zoosporangium isolates per pathogen isolate. Aggressiveness criteria were analyzed for 25 P. halstedii single zoosporangium isolates. Based on the reaction for the P. halstedii isolates to four sunflower hybrids H1–H4 varying only in their downy mildew resistance genes, there were differences in virulence spectrum in pathogen isolates. Analysis of five single zoosporangium isolates for P. halstedii isolates showed significant variability within pathogen isolate for all aggressiveness criteria but not for all pathogen isolates. The hypothesis explaining the interaction between P. halstedii and its host plant was discussed on the level of pathogenicity.     

The combined effect of color and odor on flower choice behavior of bumble bees in flower mimicry systems

Food-deceptive flowers are pollinated by animals that expecta reward but are cheated. Such plants profit from their similarityto rewarding plants and should develop signals that hinderdiscrimination. We use artificial rewarding model flowers andnonrewarding mimicking flowers that present similar visual cues. We test how additional scent cues change flower choiceof the mimic by bumble bees (Bombus terrestris) in two situations:(1) both flower types are simultaneously present and can becompared by the pollinator, and (2) both flower types are encounteredsuccessively in the absence of each other. We find that insituation 1, discrimination learning is greater if scents areused as cues for identifying the mimic, whether the mimic hasa different scent or if it is scentless while the model isscented. In situation 2, a generalization task, a scented mimicis avoided faster than a scentless one. Discrimination of themimic is poorest if it has the same scent as the model, thusdemonstrating a potential for scent mimicry, which has not yet been proved to exist among differently rewarding flowers. Thus,the best strategy for a mimic would be to have the same scentas the model, but this strategy may not be used due to evolutionaryconstraints. Alternatively, if there are several potentialmodels, then having no scent would be a better strategy thanmimicking just one of the models. In situation 1 flower discriminationby color cues is enhanced in the mere presence of scent, comparedto unscented controls, even if the scent does not provide a distinguishable cue itself. The results indicate that the presenceof scent may enhance color discrimination by improving attentiontowards visual cues and/or that combined color/odor cues maylead to better memory formation and retrieval.     

Effects of the embryonic axis and exogenous growth regulators on sunflower cotyledon storage protein mobilization

Cotyledons of sunflower seedlings ( Helianthus annuus L. cv. Giant gray stripe) expand and their protein content first rises then begins to decrease during the first three days of growth. Storage protein structures, which are visible with scanning electron microscopy, undergo modification that leads to storage protein disappearance by day 4 post-imbibition. Expansion of cotyledons detached from seeds prior to imbibition is greatly reduced, total protein levels remain high, and storage protein structures remain visible in cells of these cotyledons. Incubation of excised cotyledons in 1.0 μM benzyladenine or kinetin increases the rates of cotyledon expansion and storage protein loss to levels higher than in intact seedling cotyledons, Incubation in 10 μM indole-3-acetic acid inhibits cotyledon expansion and protein mobilization. More rapid hydrolysis of storage proteins in cotyledons of intact seedlings or detached cotyledons treated with cytokinin is further indicated in day 2 specimens by SDS-polyacrylamide gel electrophoresis. These results suggest a possible mechanism for regulation of cotyledon development by interactions of the promotive effects of cytokinin and inhibitory effects of auxin.     

Genetic distance as a predictor of heterosis and hybrid performance within and between heterotic groups in sunflower

Heterosis is significant for seed yield and is one of the driving forces behind the hybrid seed industry in cultivated sunflower (Helianthus annuus L). Heterotic groups in sunflower, if any other than the female and male inbred-line groups exist, have not been well studied or described. The primary aims of this study were to assess the utility and validity of a series of proposed heterotic groups and estimate correlations between genetic distance, heterosis, and hybrid performance for seed yield in sunflower. Fortytwo female by male heterotic group (A × R) and 81 female by female heterotic group (A × B) single-cross hybrids were grown in Corvallis, Ore., and Casselton, N.D., in 1996 and 1997. Heterosis was significant for seed yield and plant height but not for seed oil concentration and days to flowering. Genetic distances were significantly correlated with hybrid seed yield when estimated from AFLP fingerprints (G _D) (r = 0.63 for A × R and 0.79 for A × B hybrids), but not from coancestries (G _C) (r = -0.02 for A × R and 0.54 for A × B hybrids). G _D (R ² = 0.4) was a poor predictor of hybrid seed yield. The proposed heterotic groups in sunflower seem to have utility, but do not seem to be as strongly differentiated as those in corn (Zea mays L.). The highest-yielding hybrids were from the B_C× R_B heterotic pattern; however, several B_C× B_C hybrids (within-group hybrids) were among the top-yielding hybrids. The outstanding performance of certain B_C× B_C hybrids casts some doubt on the validity of the B_C group. Substantial genetic diversity seems to be present within and between heterotic groups in sunflower. Received: 1 September 1998 / Accepted: 14 September 1999     

Effects of salinity on the contents of polyamines and some other compounds in sunflower plants differing in salt tolerance

The effects of salt stress (50, 100, and 150 mM NaCl) on the levels of free, bound, and total polyamines were studied in the leaf tissues of salt-tolerant (Coban) and salt-sensitive (Sanbro) cultivars of sunflower (Helianthus annuus L.) plants grown for 15 or 25 days under salinity. The amounts of free, acid-soluble bound, and total spermine increased in leaf tissues of sunflower plants subjected to salt stress while the levels of other polyamines decreased or no significant changes occurred. The increase in some PA titers suggests their potential role in overcoming the adverse effect of salinity stress. The salt sensitivity of the sunflower plants was associated with the excessive accumulation of total polyamines in the leaf tissues of salt-sensitive cultivar (Sanbro) under saline condition. The content of other compounds such as proline, protein, and ions (Na⁺, K⁺, Cl^-, K⁺/Na⁺) in leaf tissue changed depending on salt concentration and the cultivars used.From Fiziologiya Rastenii, Vol. 52, No. 1, 2005, pp. 36–42.Original English Text Copyright © 2005 by Mutlu, Bozcuk.This article was presented by the authors in English.     

Cloning,sequence and characterization of a sunflower (Helianthus annuus L.) pathogen-induced gene showing sequence homology with auxin-induced genes from plants

The establishment of a plant-pathogen interaction involves changes in gene expressions in both organisms. To isolate Helianthus annuus genes whose expression is induced during processes of resistance to Plasmopara halstedii, a comparison of the expression pattern of healthy sunflowers was made with sunflowers infected with 2 races of P. halstedii, either virulent or avirulent, using differential display of mRNA. A full-length cDNA, HaAC1, representing a sunflower gene whose expression is enhanced during early stages of the incompatible interaction, was isolated. Different timing of RNA accumulation is observed between compatible and incompatible combinations. Sequence analysis and database search revealed significant homology with auxin-induced genes from plants. The expression of this gene, is also induced after treatment with 2,4-dichlorophenoxyacetic acid (2,4-D), salicylic acid (SA) and wounding.     

Cloning and characterization of a root sunflower peroxidase gene putatively involved in cell elongation

A cDNA-encoding a peroxidase (Helianthus annuus POX (HaPOX)1) was isolated and characterized from the roots of sunflower seedlings. This gene exhibited homology with other peroxidases from several higher-plant species, and its expression in the root growth was particularly abundant during cell expansion. To elucidate the precise functions of HaPOX1 in sunflower root, we examined its expression pattern in response to several plant growth regulators. Expression of HaPOX1 is down-regulated by abscisic acid (ABA), whereas indole-3-acetic acid (IAA) induced its expression. These results suggest that HaPOX1 expression is differentially regulated by phytohormonal components of signaling cascades. Since IAA appears to participate in the regulation of HaPOX1 expression, we postulate that the peroxidase encoded by HaPOX1 may be involved in the reactions that promote cell elongation during the early stage of root growth.