BackgroundHoney bees (Apis mellifera) provide a principal example of diphenic development. Excess feeding of female larvae results in queens (large reproductives). Moderate diet yields workers (small helpers). The signaling pathway that links provisioning to female developmental fate is not understood, yet we reasoned that it could include TOR (target of rapamycin), a nutrient- and energy-sensing kinase that controls organismal growth.Methodology/Principal FindingsHere, the role of Apis mellifera TOR (amTOR) in caste determination is examined by rapamycin/FK506 pharmacology and RNA interference (RNAi) gene knockdown. We show that in queen-destined larvae, the TOR inhibitor rapamycin induces the development of worker characters that are blocked by the antagonist FK506. Further, queen fate is associated with elevated activity of the Apis mellifera TOR encoding gene, amTOR, and amTOR gene knockdown blocks queen fate and results in individuals with worker morphology.Conclusions/SignificanceA much-studied insect dimorphism, thereby, can be governed by the TOR pathway. Our results present the first evidence for a role of TOR in diphenic development, and suggest that adoption of this ancestral nutrient-sensing cascade is one evolutionary pathway for morphological caste differentiation in social insects.

SUMMARY Nutrient sensitive insulin-like peptides (ILPs) have profound effects on invertebrate metabolism, nutrient storage, fertility and aging. Many insects transcribe ILPs in specialized neurosecretory cells at changing levels correlated with life history. However, the major site of insect metabolism and nutrient storage is not the brain, but rather the fat body, where functions of ILP expression are rarely studied and poorly understood. Fat body is analogous to mammalian liver and adipose tissue, with nutrient stores that often correlate with behavior. We used the honey bee (Apis mellifera), an insect with complex behavior, to test whether ILP genes in fat body respond to experimentally induced changes of behavioral physiology. Honey bee fat body influences endocrine state and behavior by secreting the yolk protein precursor vitellogenin (Vg), which suppresses lipophilic juvenile hormone and social foraging behavior. In a two-factorial experiment, we used RNA interference (RNAi)-mediated vg gene knockdown and amino acid nutrient enrichment of hemolymph (blood) to perturb this regulatory module. We document factor-specific changes in fat body ilp1 and ilp2 mRNA, the bee's ILP-encoding genes, and confirm that our protocol affects social behavior. We show that ilp1 and ilp2 are regulated independently and differently and diverge in their specific expression-localization between fat body oenocyte and trophocyte cells. Insect ilp functions may be better understood by broadening research to account for expression in fat body and not only brain.

In honeybee colonies, food collection is performed by a group of mostly sterile females called workers. After an initial nest phase, workers begin foraging for nectar and pollen, but tend to bias their collection towards one or the other. The foraging choice of honeybees is influenced by vitellogenin (vg), an egg-yolk precursor protein that is expressed although workers typically do not lay eggs. The forager reproductive ground plan hypothesis (RGPH) proposes an evolutionary path in which the behavioural bias toward collecting nectar or pollen on foraging trips is influenced by variation in reproductive physiology, such as hormone levels and vg gene expression. Recently, the connections between vg and foraging behaviour were challenged by Oldroyd and Beekman (2008), who concluded from their study that the ovary, and especially vg, played no role in foraging behaviour of bees. We address their challenge directly by manipulating vg expression by RNA interference-(RNAi) mediated gene knockdown in two honeybee genotypes with different foraging behaviour and reproductive physiology. We show that the effect of vg on the food-loading decisions of the workers occurs only in the genotype where timing of foraging onset (by age) is also sensitive to vg levels. In the second genotype, changing vg levels do not affect foraging onset or bias. The effect of vg on workers' age at foraging onset is explained by the well-supported double repressor hypothesis (DHR), which describes a mutually inhibitory relationship between vg and juvenile hormone (JH) -an endocrine factor that influences development, reproduction, and behaviour in many insects. These results support the RGPH and demonstrate how it intersects with an established mechanism of honeybee behavioural control. KeywordsApis mellifera; juvenile hormone; reproductive ground plan hypothesis; RNA interference; social foraging; vitellogenin yolk protein Recent advances in functional genomics have enabled new insights and the direct testing of hypotheses about the molecular regulation of behaviour. The eusocial honeybee has emerged as a model organism for the study of molecular mechanisms that can influence complex social behaviour (Grozinger and Robinson 2007;Marco Antonio et al 2008). Worker bees are

As species evolve, they become adapted to their local environments. Detecting the genetic signature of selection and connecting that to the phenotype of the organism, however, is challenging. Here we report using an integrative approach that combines DNA sequencing with structural biology analyses to assess the effect of selection on residues in the mitochondrial DNA of the two species of African elephants. We detected evidence of positive selection acting on residues in complexes I and V, and we used homology protein structure modeling to assess the effect of the biochemical properties of the selected residues on the enzyme structure. Given the role these enzymes play in oxidative phosphorylation, we propose that the selected residues may contribute to the metabolic adaptation of forest and savanna elephants to their unique habitats.

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