Pleiotropy, epistasis and new QTL: the genetic architecture of honey bee foraging behavior.

The regulation of division of labor in social insects, particularly in the honey bee (Apis mellifera L.), has received considerable attention from a number of biological subdisciplines, including quantitative and behavioral genetics, because of the high complexity of the behavioral traits involved. The foraging choices of honey bee workers can be accurately quantified, and previous studies have made the foraging behavior of honey bees one of the best studied naturally occurring behavioral phenotypes. Three quantitative trait loci (QTL) have been identified that influence a set of foraging variables, including the concentration of nectar collected and the amount of pollen and nectar brought back to the hive. This study extends previous genetic investigations and represents the most comprehensive investigation of the genetic architecture of these foraging variables. We examined the effects of markers for the three established QTL and for one further candidate gene (Amfor), in two reciprocal backcross populations. These populations were also used to carry out two new QTL mapping studies, with over 400 Amplified Fragment Length Polymorphism (AFLP) markers in each. We detected a variety of effects of the genetic markers for the established QTL and the candidate gene, which were mostly epistatic in nature. A few new QTL could be detected with a variety of mapping techniques. Our results add complexity to the genetic architecture of the foraging behavior of the honey bee. Specifically, we support the hypotheses that pln1, pln2, pln3, and Amfor are involved in the regulation of foraging behavior in the honey bee and add some new factors that deserve further study in the future.

Effect of pheromones, hormones, and handling on sucrose response thresholds of honey bees (Apis mellifera L.).

The responsiveness of bees to sucrose is an important indicator of honey bee foraging decisions. Correlated with sucrose responsiveness is forage choice behavior, age of first foraging, and conditioned learning response. Pheromones and hormones are significant components in social insect systems associated with the regulation of colony-level and individual foraging behavior. Bees were treated to different exposure regimes of queen and brood pheromones and their sucrose responsiveness measured. Bees reared with queen or brood pheromone were less responsive than controls. Our results suggest responsiveness to sucrose is a physiologically, neuronally mediated response. Orally administered octopamine significantly reduced sucrose response thresholds. Change in response to octopamine was on a time scale of minutes. The greatest separation between octopamine treated and control bees occurred 30 min after feeding. There was no significant sucrose response difference to doses ranging from 0.2 mug to 20 mug of octopamine. Topically applied methoprene significantly increased sucrose responsiveness. Handling method significantly affected sucrose responsiveness. Bees that were anesthetized by chilling or CO(2) treatment were significantly more responsive than control bees 30 min after handling. Sixty minutes after handling there were no significant treatment differences. We concluded that putative stress effects of handling were blocked by anesthetic.

Modulation of sucrose response thresholds in honey bees (Apis mellifera L.): influence of genotype, feeding, and foraging experience.

The perception of sugar is important to honey bees for making foraging decisions. We measured bees' perception by determining what concentration of sucrose touched to the antennae elicited the proboscis extension response (response threshold). A low response threshold (extension at low concentration) suggests a high perceptual value of sucrose. and vice versa. Perception of sucrose solutions differed between two artificially selected genotypic strains and was modulated by the bees' recent feeding experiences. Bees offered 10%, 30%, or 50% sucrose solutions in small cages overnight, and in large flight-cages or free-flying in the field for several days, had subsequent response thresholds positively correlated to the concentration offered. Empty bees, whether they were nectar, water or pollen foragers, dancers or non-dancers, had a significantly lower threshold than loaded bees. Crop volume affected response thresholds directly and independently of sucrose concentration. We interpret these findings as multiple mechanisms that operate in different time scales, modulating perception of sucrose. Changes occurred in the time scale of evolutionary processes as demonstrated by genotypic differences. Changes with foraging experience occur in hours or minutes while effects of crop filling are instantaneous.

Selection on worker honeybee responses to queen pheromone (Apis mellifera L.).

Disruptive selection for responsiveness to queen mandibular gland pheromone (QMP) in the retinue bioassay resulted in the production of high and low QMP responding strains of honeybees (Apis mellifera L.). Strains differed significantly in their retinue response to QMP after one generation of selection. By the third generation the high strain was on average at least nine times more responsive than the low strain. The strains showed seasonal phenotypic plasticity such that both strains were more responsive to the pheromone in the spring than in the fall. Directional selection for low seasonal variation indicated that phenotypic plasticity was an additional genetic component to retinue response to QMP. Selection for high and low retinue responsiveness to QMP was not an artifact of the synthetic blend because both strains were equally responsive or non-responsive to whole mandibular gland extracts compared with QMP. The use of these strains clearly pointed to an extra-mandibular source of retinue pheromones (Pankiw et al. 1995; Slessor et al. 1998; Keeling et al. 1999).

The effect of genotype, age, sex, and caste on response thresholds to sucrose and foraging behavior of honey bees (Apis mellifera L.).

Bees derived from artificially selected high- and low-pollen-hoarding strains were tested for their proboscis extension reflex response to water and varying sucrose concentrations. High-strain bees had a lower response threshold to sucrose than low-strain bees among pre-foragers, foragers, queens and drones. Pre-foraging low-strain workers showed ontogenetic changes in their response threshold to sucrose which was inversely related to age. High-strain foragers were more likely to return with loads of water compared to low-strain foragers. Whereas low-strain foragers were more likely to return with loads of nectar. Low-strain nectar foragers collected nectar with significantly higher sucrose concentrations than did the high-strain nectar foragers. Alternatively, low-strain foragers were more likely to return empty compared to high-strain foragers. These studies demonstrate how a genotypically varied sensory-physiological process, the perception of sucrose, are associated with a division of labor for foraging.

Queen mandibular gland pheromone influences worker honey bee (Apis mellifera L.) foraging ontogeny and juvenile hormone titers.

SYNTHETIC QUEEN MANDIBULAR GLAND PHEROMONE (QMP) WAS APPLIED TO HONEY BEE COLONIES TO TEST TWO HYPOTHESES: (i) QMP acts like a primer pheromone in the regulation of age-related division of labor, and (ii) this primer effect, if present, varies in three strains of workers that show genetically-based differences in their retinue attraction response to QMP (a pheromone releaser effect). Strains of workers that were high, or low in their response to QMP in a laboratory bioassay, as well as unselected 'wild-type' workers, were fostered in queenright colonies with or without supplemental QMP. Effects of QMP on foraging ontogeny and juvenile hormone III (JH) blood titers in worker honey bees were measured. Bees in QMP-supplemented colonies showed significant delays in foraging ontogeny, and foraging activity was reduced. They also had significantly lower JH titers, although the titer curves were somewhat atypical. There were no differences in foraging ontogeny or JH titers among the three strains. We conclude that (i) QMP can delay the ontogeny of foraging by some mechanism that suppresses JH production, (ii) this QMP primer response is independent of the retinue releaser response, and (iii) QMP can play an important role in regulating division of labour.

Mandibular gland components of european and africanized honey bee queens (Apis mellifera L.).

The composition of the five-component honey bee queen mandibular gland pheromone (QMP) of mated European honey bee queens was compared to those of virgin and drone-laying (i.e., laying only haploid unfertilized eggs that develop into males), European queens and Africanized mated queens. QMP of mated European queens showed significantly greater quantities of individual components than all queen types compared, except for a significantly greater quantity of 9-hydroxy-(E)-2-decenoic acid (9-HDA) found in Africanized queens. Glands of European drone-laying queens contained quantities intermediate between virgin and mated queens, reflecting their intermediate reproductive state and age. QMP ontogeny shifts from a high proportion of 9-keto-(E)-2-decenoic acid (ODA) in young unmated queens to roughly equal proportions of ODA and 9-HDA in mated queens. A biosynthetic shift occurs after mating that results in a greater proportion of 9-HDA, methylp-hydroxybenzoate (HOB), and 4-hydroxy-3-methoxyphenylethanol (HVA) production, accompanied by a decreased proportion of ODA. Africanized QMP proportions of ODA and 9-HDA were significantly different from European queens. A quantitative definition of a "queen equivalent" of QMP is proposed for the various queen types, and a standard queen equivalent for mated European honeybee queen mandibular gland pheromone is adopted as 200µg ODA, 80µg 9-HDA, 20µg HOB, and 2 µg HVA.