More than mesolectic: Characterizing the nutritional niche of Osmia cornifrons.

Characterizing the nutritional needs of wild bee species is an essential step to better understanding bee biology and providing suitable supplemental forage for at-risk species. Here, we aim to characterize the nutritional needs of a model solitary bee species, Osmia cornifrons (Radoszkowski), by using dietary protein-to-lipid ratio (P:L ratio) as a proxy for nutritional niche and niche breadth. We first identified the mean target P:L ratio (~3.02:1) and P:L collection range (0.75-6.26:1) from pollen provisions collected across a variety of sites and time points. We then investigated the P:L tolerance range of larvae by rearing bees in vitro on a variety of diets. Multifloral and single-source pollen diets with P:L ratios within the range of surveyed provisions did not always support larval development, indicating that other dietary components such as plant secondary compounds and micronutrients must also be considered in bee nutritional experiments. Finally, we used pollen metabarcoding to identify pollen from whole larval provisions to understand how much pollen bees used from plants outside of their host plant families to meet their nutritional needs, as well as pollen from individual forager bouts, to observe if bees maintained strict floral constancy or visited multiple plant genera per foraging bout. Whole larval provision surveys revealed a surprising range of host plant pollen use, ranging from ~5% to 70% of host plant pollen per provision. Samples from individual foraging trips contained pollen from multiple genera, suggesting that bees are using some form of foraging decision making. Overall, these results suggest that O. cornifrons have a wide nutritional niche breadth, but while pollen P:L ratio tolerance is broad, a tolerable P:L ratio alone is not enough to create a quality diet for O. cornifrons, and the plant species that make up these diets must also be carefully considered.

Beyond conflict: Kinship theory of intragenomic conflict predicts individual variation in altruistic behaviour.

Behavioural variation is essential for animals to adapt to different social and environmental conditions. The Kinship Theory of Intragenomic Conflict (KTIC) predicts that parent-specific alleles can support different behavioural strategies to maximize allele fitness. Previous studies, including in honey bees (Apis mellifera), supported predictions of the KTIC for parent-specific alleles to promote selfish behaviour. Here, we test the KTIC prediction that for altruism-promoting genes (i.e. those that promote behaviours that support the reproductive fitness of kin), the allele with the higher altruism optimum should be selected to be expressed while the other is silenced. In honey bee colonies, workers act altruistically when tending to the queen by performing a 'retinue' behaviour, distributing the queen's mandibular pheromone (QMP) throughout the hive. Workers exposed to QMP do not activate their ovaries, ensuring they care for the queen's brood instead of competing to lay unfertilized eggs. Due to the haplodiploid genetics of honey bees, the KTIC predicts that response to QMP is favoured by the maternal genome. We report evidence for parent-of-origin effects on the retinue response behaviour, ovarian development and gene expression in brains of worker honey bees exposed to QMP, consistent with the KTIC. Additionally, we show enrichment for genes with parent-of-origin expression bias within gene regulatory networks associated with variation in bees' response to QMP. Our study demonstrates that intragenomic conflict can shape diverse social behaviours and influence expression patterns of single genes as well as gene networks.

Examining parent-of-origin effects on transcription and RNA methylation in mediating aggressive behavior in honey bees (Apis mellifera).

Conflict between genes inherited from the mother (matrigenes) and the father (patrigenes) is predicted to arise during social interactions among offspring if these genes are not evenly distributed among offspring genotypes. This intragenomic conflict drives parent-specific transcription patterns in offspring resulting from parent-specific epigenetic modifications. Previous tests of the kinship theory of intragenomic conflict in honey bees (Apis mellifera) provided evidence in support of theoretical predictions for variation in worker reproduction, which is associated with extreme variation in morphology and behavior. However, more subtle behaviors - such as aggression - have not been extensively studied. Additionally, the canonical epigenetic mark (DNA methylation) associated with parent-specific transcription in plant and mammalian model species does not appear to play the same role as in honey bees, and thus the molecular mechanisms underlying intragenomic conflict in this species is an open area of investigation. Here, we examined the role of intragenomic conflict in shaping aggression in honey bee workers through a reciprocal cross design and Oxford Nanopore direct RNA sequencing. We attempted to probe the underlying regulatory basis of this conflict through analyses of parent-specific RNA m6A and alternative splicing patterns. We report evidence that intragenomic conflict occurs in the context of honey bee aggression, with increased paternal and maternal allele-biased transcription in aggressive compared to non-aggressive bees, and higher paternal allele-biased transcription overall. However, we found no evidence to suggest that RNA m6A or alternative splicing mediate intragenomic conflict in this species.

Tissue-specific transcriptional patterns underlie seasonal phenotypes in honey bees (Apis mellifera).

Faced with adverse conditions, such as winter in temperate regions or hot and dry conditions in tropical regions, many insect species enter a state of diapause, a period of dormancy associated with a reduction or arrest of physical activity, development and reproduction. Changes in common physiological pathways underlie diapause phenotypes in different insect species. However, most transcriptomic studies of diapause have not simultaneously evaluated and compared expression patterns in different tissues. Honey bees (Apis mellifera) represent a unique model system to study the mechanisms underpinning diapause-related phenotypes. In winter, honey bees exhibit a classic diapause phenotype, with reduced metabolic activity, increased physiological nutritional resources and altered hormonal profiles. However, winter bees actively heat their colony by vibrating their wing muscles; thus, this tissue is not quiescent. Here, we evaluated the transcriptional profiles of flight muscle tissue and fat body tissue (involved in nutrient storage, metabolism and immune function) of winter bees. We also evaluated two behavioural phenotypes of summer bees: nurses, which exhibit high nutritional stores and low flight activity, and foragers, which exhibit low nutritional stores and high flight activity. We found winter bees and nurses have similar fat body transcriptional profiles, whereas winter bees and foragers have similar flight muscle transcriptional profiles. Additionally, differentially expressed genes were enriched in diapause-related gene ontology terms. Thus, honey bees exhibit tissue-specific transcriptional profiles associated with seasonal phenotypes, laying the groundwork for future studies evaluating the mechanisms, evolution and consequences of this tissue-specific regulation.