Differential ability of three bee species to move genes via pollen.

Since the release of genetically engineered (GE) crops, there has been increased concern about the introduction of GE genes into non-GE fields of a crop and their spread to feral or wild cross-compatible relatives. More recently, attention has been given to the differential impact of distinct pollinators on gene flow, with the goal of developing isolation distances associated with specific managed pollinators. To examine the differential impact of bee species on gene movement, we quantified the relationship between the probability of getting a GE seed in a pod, and the order in which a flower was visited, or the cumulative distance traveled by a bee in a foraging bout. We refer to these relationships as 'seed curves' and compare these seeds curves among three bee species. The experiments used Medicago sativa L. plants carrying three copies of the glyphosate resistance (GR) allele as pollen donors (M. sativa is a tetraploid), such that each pollen grain carried the GR allele, and conventional plants as pollen recipients. Different foraging metrics, including the number of GR seeds produced over a foraging bout, were also quantified and contrasted among bee species. The lowest number of GR seeds set per foraging bout, and the GR seeds set at the shortest distances, were produced following leafcutting bee visits. In contrast, GR seeds were found at the longest distances following bumble bee visits. Values for honey bees were intermediate. The ranking of bee species based on seed curves correlated well with field-based gene flow estimates. Thus, differential seed curves of bee species, which describe patterns of seed production within foraging bouts, translated into distinct abilities of bee species to move genes at a landscape level. Bee behavior at a local scale (foraging bout) helps predict gene flow and the spread of GE genes at the landscape scale.

Bee species visiting Medicago sativa differ in pollen deposition curves with consequences for gene flow.

PREMISE: Pollinator foraging behavior can influence pollen dispersal and gene flow. In many plant species a pollinator trips a flower by applying pressure to release its sexual organs. We propose that differences in tripping rate among grooming pollinators could generate distinct pollen deposition curves, the pattern of pollen deposition over successive flowers visited. This study compares the pollen deposition curves of two grooming pollinators, a social bumble bee and a solitary leafcutting bee, with distinct tripping rates on Medicago sativa flowers. We predict a steeper deposition curve for pollen moved by leafcutting bees, the pollinator with the higher tripping rate. METHODS: Medicago sativa plants carrying a gene (GUS) whose product is easily detected by staining, were used as pollen donors. After visiting the GUS plants, a bee was released on a linear array of conventional M. sativa plants. The number of GUS pollen grains deposited over successive flowers visited or over cumulative distances was examined. Distinct mixed effect Poisson regression models, illustrating different rates of decay in pollen deposition, were fitted to the pollen data for each bee species. RESULTS: Pollen decay was steeper for leafcutting bees relative to bumble bees for both models of flowers visited and cumulative distance, as predicted by their higher tripping rate. CONCLUSIONS: This is the first report of a difference in pollen deposition curves between two bee species, both grooming pollinators. Such differences could lead to distinct impacts of bee species on gene flow, genetic differentiation, introgression, and ultimately speciation.

Linking the foraging behavior of three bee species to pollen dispersal and gene flow.

Foraging behaviors that impact gene flow can guide the design of pollinator strategies to mitigate gene flow. Reduced gene flow is expected to minimize the impact of genetically engineered (GE) crops on feral and natural populations and to facilitate the coexistence of different agricultural markets. The goal of this study is to link foraging behavior to gene flow and identify behaviors that can help predict gene flow for different bee species. To reach this goal, we first examined and compared the foraging behaviors of three distinct bee species, the European honey bee, Apis mellifera L., the common eastern bumble bee, Bombus impatiens Cr., and the alfalfa leafcutting bee, Megachile rotundata F., foraging on Medicago sativa flowers. Each foraging behavior investigated differed among bee species. Both social bees exhibited directionality of movement and had similar residence, in contrast to the random movement and shorter residence of the solitary bee. Tripping rate and net distance traveled differed among the three bee species. We ranked each behavior among bee species and used the relative ranking as gene flow predictor before testing the predictions against empirical gene flow data. Tripping rate and net distance traveled, but not residence, predicted relative gene dispersal among bee species. Linking specific behaviors to gene flow provides mechanisms to explain differences in gene flow among bee species and guides the development of management practices to reduce gene flow. Although developed in one system, the approach developed here can be generalized to different plant/pollinator systems.

The role of pollinators in maintaining variation in flower colour in the Rocky Mountain columbine, Aquilegia coerulea.

BACKGROUND AND AIMS: Flower colour varies within and among populations of the Rocky Mountain columbine, Aquilegia coerulea, in conjunction with the abundance of its two major pollinators, hawkmoths and bumble-bees. This study seeks to understand whether the choice of flower colour by these major pollinators can help explain the variation in flower colour observed in A. coerulea populations. METHODS: Dual choice assays and experimental arrays of blue and white flowers were used to determine the preference of hawkmoths and bumble-bees for flower colour. A test was made to determine whether a differential preference for flower colour, with bumble-bees preferring blue and hawkmoths white flowers, could explain the variation in flower colour. Whether a single pollinator could maintain a flower colour polymorphism was examined by testing to see if preference for a flower colour varied between day and dusk for hawkmoths and whether bumble-bees preferred novel or rare flower colour morphs. KEY RESULTS: Hawkmoths preferred blue flowers under both day and dusk light conditions. Naïve bumble-bees preferred blue flowers but quickly learned to forage randomly on the two colour morphs when similar rewards were presented in the flowers. Bees quickly learned to associate a flower colour with a pollen reward. Prior experience affected the choice of flower colour by bees, but they did not preferentially visit novel flower colours or rare or common colour morphs. CONCLUSIONS: Differences in flower colour preference between the two major pollinators could not explain the variation in flower colour observed in A. coerulea. The preference of hawkmoths for flower colour did not change between day and dusk, and bumble-bees did not prefer a novel or a rare flower colour morph. The data therefore suggest that factors other than pollinators may be more likely to affect the flower colour variation observed in A. coerulea.

Impact of insect pollinator group and floral display size on outcrossing rate.

Despite the strong influence of pollination ecology on the evolution of selfing, we have little information on how distinct groups of insect pollinators influence outcrossing rate. However, differences in behavior between pollinator groups could easily influence how each group affects outcrossing rate. We examined the influence of distinct insect pollinator groups on outcrossing rate in the rocky mountain columbine, Aquilegia coerulea. The impact of population size, plant density, size of floral display, and herkogamy (spatial separation between anthers and stigmas) on outcrossing rate was also considered as these variables were previously found to affect outcrossing rate in some plant species. We quantified correlations between all independent variables and used simple and two-factor regressions to determine direct and indirect impact of each independent variable on outcrossing rate. Outcrossing rate increased significantly with hawkmoth abundance but not with the abundance of any of the other groups of floral visitors, which included bumblebees, solitary bees, syrphid flies, and muscidae. Outcrossing rate was also significantly affected by floral display size and together, hawkmoth abundance and floral display size explained 87% of the variation in outcrossing rate. None of the other independent variables directly affected the outcrossing rate. This is the first report of a significant impact of pollinator type on outcrossing rate. Hawkmoths did not visit fewer flowers per plant relative to other pollinator groups but preferred visiting female-phase flowers first on a plant. Both the behavior of pollinators and floral display size affected outcrossing rate via their impact on the level of geitonogamous (among flower) selfing. Given that geitonogamous selfing is never advantageous, the variation in outcrossing rate and maintenance of mixed mating systems in populations of A. coerulea may not require an adaptive explanation.