Characterization of 14 polymorphic microsatellite loci for the facultatively eusocial sweat bee Halictus rubicundus (Hymenoptera, Halictidae) and their variability in related species

Sweat bees display considerable variation in social organization and a few species, such as Halictus rubicundus, are even facultatively eusocial. Fourteen polymorphic, unlinked microsatellite loci were isolated from H. rubicundus and characterized in 45 females. The number of alleles per locus ranged from two to 18 (mean 10.1), observed heterozygosity ranged from 0.24 to 0.98 (mean 0.71) and expected heterozygosity ranged from 0.24 to 0.98 (mean 0.70). Six or more loci cross-amplified in four other sweat bees. These loci will be useful for the study of social evolution and population genetic structure in H. rubicundus and many other sweat bees.     

Genetic differentiation across the social transition in a socially polymorphic sweat bee,Halictus rubicundus

Eusociality is widely considered a major evolutionary transition. The socially polymorphic sweat bee Halictus rubicundus, solitary in cooler regions of its Holarctic range and eusocial in warmer parts, is an excellent model organism to address this transition, and specifically the question of whether sociality is associated with a strong barrier to gene flow between phenotypically divergent populations. Mitochondrial DNA (COI) from specimens collected across the British Isles, where both solitary and social phenotypes are represented, displayed limited variation, but placed all specimens in the same European lineage; haplotype network analysis failed to differentiate solitary and social lineages. Microsatellite genetic variability was high and enabled us to quantify genetic differentiation among populations and social phenotypes across Great Britain and Ireland. Results from conceptually different analyses consistently showed greater genetic differentiation between geographically distant populations, independently of their social phenotype, suggesting that the two social forms are not reproductively isolated. A landscape genetic approach revealed significant isolation by distance (Mantel test r = 0.622, P < 0.001). The Irish Sea acts as physical barrier to gene flow (partial Mantel test r = 0.453, P < 0.01), indicating that geography, rather than expression of solitary or social behaviour (partial Mantel test r = −0.238, P = 0.053), had a significant effect on the genetic structure of H. rubicundus across the British Isles. Although we cannot reject the hypothesis of a genetic underpinning to differences in solitary and eusocial phenotypes, our data clearly demonstrate a lack of reproductive isolation between the two social forms.     

Colony structure, provisioning and sex allocation in the sweat bee Halictus ligatus (Hymenoptera: Halictidae)

Population and colony-level sex allocation and nest productivity in the eusocial sweat bee Halictus ligatus Say were studied by excavating nests during one season. The emphasis was on measuring the provision masses, which differ in size and shape depending on the sex of the egg to be laid on them (male-producing provision masses are smaller and more or less round, whereas gyne-producing provision masses are larger and 'loaf-shaped). The primary aim of this study was to test theoretical predictions about female-bias of the sex ratio in the summer brood, both on the population level and on the colony level.
The overall sex ratio of the summer brood was moderately biased towards gynes. A significant positive correlation between the overall size of provision masses (as an estimate for the degree of female bias of the nest sex ratio) and the number of eusocial workers was found. This relationship further improved in partial analyses in which the provision mass weights were adjusted for sampling date, removing the effect of protandry. Foundress size, however, had no effect on the second brood provision masses and neither was there an effect of worker number on the size of gynes and males separately. In the first brood only the size of the foundress had a consistently positive effect on the size of the provision masses and on the size of the emerging daughter workers.
The observed increase of female bias in the nest sex ratio with increasing numbers of eusocial worker bees conforms to optimization predictions following from kin-selection theory.     

Socially induced brain development in a facultatively eusocial sweat bee Megalopta genalis (Halictidae)

Changes in the relative size of brain regions are often dependent on experience and environmental stimulation, which includes an animal''s social environment. Some studies suggest that social interactions are cognitively demanding, and have examined predictions that the evolution of sociality led to the evolution of larger brains. Previous studies have compared species with different social organizations or different groups within obligately social species. Here, we report the first intraspecific study to examine how social experience shapes brain volume using a species with facultatively eusocial or solitary behaviour, the sweat bee Megalopta genalis. Serial histological sections were used to reconstruct and measure the volume of brain areas of bees behaving as social reproductives, social workers, solitary reproductives or 1-day-old bees that are undifferentiated with respect to the social phenotype. Social reproductives showed increased development of the mushroom body (an area of the insect brain associated with sensory integration and learning) relative to social workers and solitary reproductives. The gross neuroanatomy of young bees is developmentally similar to the advanced eusocial species previously studied, despite vast differences in colony size and social organization. Our results suggest that the transition from solitary to social behaviour is associated with modified brain development, and that maintaining dominance, rather than sociality per se, leads to increased mushroom body development, even in the smallest social groups possible (i.e. groups with two bees). Such results suggest that capabilities to navigate the complexities of social life may be a factor shaping brain evolution in some social insects, as for some vertebrates.     

Lasioglossum (Dialictus) tenax (Sandhouse) (Hymenoptera: Halictidae) as a solitary sweat bee

Summary Nine nests ofLasioglossum (Dialictus) tenax were excavated near Calgary, Alberta, Canada over a time period encompassing the entire brood production period in 1988. Each nest contained a maximum of one active adult female, nest productivity peaked in mid July, protandry was noted and no significant size difference between foundresses and the earlier emerging females was detected. These data suggest that this species is solitary. These results are compared with data for the sympatrically nesting eusocial speciesL. (D.) laevissimum.     

Novel antimicrobial peptides from the venom of the eusocial bee Halictus sexcinctus (Hymenoptera: Halictidae) and their analogs

Two novel antimicrobial peptides, named halictines, were isolated from the venom of the eusocial bee Halictus sexcinctus. Their primary sequences were established by ESI-QTOF mass spectrometry, Edman degradation and enzymatic digestion as Gly-Met-Trp-Ser-Lys-Ile-Leu-Gly-His-Leu-Ile-Arg-NH₂ (HAL-1), and Gly-Lys-Trp-Met-Ser-Leu-Leu-Lys–His-Ile-Leu-Lys-NH₂ (HAL-2). Both peptides exhibited potent antimicrobial activity against Gram-positive and Gram-negative bacteria but also noticeable hemolytic activity. The CD spectra of HAL-1 and HAL-2 measured in the presence of trifluoroethanol or SDS showed ability to form an amphipathic α-helical secondary structure in an anisotropic environment such as bacterial cell membrane. NMR spectra of HAL-1 and HAL-2 measured in trifluoroethanol/water confirmed formation of helical conformation in both peptides with a slightly higher helical propensity in HAL-1. Altogether, we prepared 51 of HAL-1 and HAL-2 analogs to study the effect of such structural parameters as cationicity, hydrophobicity, α-helicity, amphipathicity, and truncation on antimicrobial and hemolytic activities. The potentially most promising analogs in both series are those with increased net positive charge, in which the suitable amino acid residues were replaced by Lys. This improvement basically relates to the increase of antimicrobial activity against pathogenic Pseudomonas aeruginosa and to the mitigation of hemolytic activity.     

Phylogeny of the bee genus Agapostemon (Hymenoptera: Halictidae)

Abstract. A phylogeny for genus Agapostemon (Hymenoptera: Halictidae) is reconstructed from morphological characters, using all species, with four representatives of Agapostemonini as outgroup taxa. Parsimony analysis using 150 unordered characters resulted in eighteen equally parsimonious trees, a strict consensus of which is highly resolved for Agapostemon. Based on this analysis, Agapostemonoides Roberts & Brooks is returned to generic status, and a new subgenus is described: Notagapostemon, which includes the rhopalocerus, heterurus, atrocaeruleus, erebus, kohliellus, nasutus and intermedius groups of Moure & Hurd. Species groups are proposed for Notagapostemon and revised in A. (Agapostemon). Agapostemon cockerelli Crawford is synonymized with A. obliquus (Provancher), A. epichryseus (Morelos) with A. leunculus Vachal, A. inca Roberts with A. heterurus Cockerell, A. hispaniolicus Roberts female with A. centratus (Vachal) and A. hispaniolicus Roberts male with A. alayoi Roberts. Optimization of known behaviours suggests communal ancestry for the genus and infers the prevalence of this behaviour in Agapostemon. Optimization of geographical range suggests a Central American or Caribbean origin of Agapostemon, with one invasion of North America and multiple invasions of South America and the Caribbean. One invasion of the Caribbean suggests arrival from North America via Florida.     

Characterization of microsatellite loci from the solitary sweat bees Lasioglossum leucozonium and Lasioglossum oenotherae (Hymenoptera,Halictidae)

Microsatellite loci were isolated from two solitary sweat bees: the polylectic Lasioglossum leucozonium (10 loci) and the oligolectic Lasioglossum oenotherae (9 loci) (Hymenoptera, Halictidae). All loci were polymorphic with high observed heterozygosities (0.07–0.75 for L. leucozonium; 0.06–0.92 for L. oenotherae). These loci will be used to study the consequences of diet specialization on the population and conservation genetics of bees.     

The population genetics of a solitary oligolectic sweat bee, Lasioglossum (Sphecodogastra) oenotherae (Hymenoptera: Halictidae)

Strong evidence exists for global declines in pollinator populations. Data on the population genetics of solitary bees, especially diet specialists, are generally lacking. We studied the population genetics of the oligolectic bee Lasioglossum oenotherae, a specialist on the pollen of evening primrose (Onagraceae), by genotyping 455 females from 15 populations across the bee's North American range at six hyper-variable microsatellite loci. We found significant levels of genetic differentiation between populations, even at small geographic scales, as well as significant patterns of isolation by distance. However, using multilocus genotype assignment tests, we detected 11 first-generation migrants indicating that L. oenotherae's sub-populations are experiencing ongoing gene flow. Southern populations of L. oenotherae were significantly more likely to deviate from Hardy-Weinberg equilibrium and from genotypic equilibrium, suggesting regional differences in gene flow and/or drift and inbreeding. Short-term N(e) estimated using temporal changes in allele frequencies in several populations ranged from approximately 223 to 960. We discuss our findings in terms of the conservation genetics of specialist pollinators, a group of considerable ecological importance.     

Proventricular structure in the bee tribe Augochlorini (Hymenoptera: Halictidae)

Characters of the digestive tract have received little attention in modern phylogenetic analyses regarding relationships among bees, in part because studies on the internal morphology of bees generally concentrate on physiological and behavioural aspects. This paper presents a comparative study of the proventricular structure, analysed with scanning electronic microscopy, in bees of the tribe Augochlorini. Eleven species of Augochlorini were analysed as the ingroup, versus one each of Halictini and Caenohalictini, and two of Crabronidae as outgroups. The presence of a long columnar proventricular fold is an apomorphy for Augochlorini, whereas a proventricular fold with a keel-shaped structure at the moving lip level is an apomorphy for Augochlorina. Together these data corroborate the monophyly of Augochlorini and Augochlorina, respectively.     

The Biology of a Subtropical Population of Halictus ligatus Say (Hymenoptera; Halictidae)

The ability of Manx shearwaters (Puffinus puffinus) to locate their nesting burrows at night was investigated using observation and experiments. Shearwaters walking to their burrows did so at random with respect to the wind direction, and did not vocalize, suggesting that visual cues are important to successful burrow homing. Experiments to test whether vision, audition or olfaction functioned in guiding birds back to burrows supported this conclusion.     

A review of the bee genus Augochlorella (Hymenoptera: Halictidae: Augochlorini)

Abstract. The New World halictid bee genus Augochlorella (Augochlorini) is revised. Sixteen species are recognized, with five described as new: Augochlorella acarinata sp. n., A. una sp. n., A. meridionalis sp. n., A. stenothoracica sp. n. from South America, and A. karankawa sp. n. from U.S.A. The following new synonymies are proposed: Augochlorella michaelis (Vachal) with A. urania (Smith), A. edendata Michener with A. comis (Vachal); A. striata (Provancher) with Augochlorella aurata (Smith) and A. neglectula maritima Ordway with A. neglectula (Cockerell). The female of A. tredecim (Vachal) and the male of A. iopoecila Moure are described for the first time. Keys to the species are provided. Cladistic analysis of adult morphological characters corroborates Engel's hypotheses of monophyly of Augochlorella phylogenetic relationships with related genera as follow: (Augochlorella ((Ceratalictus, Pereirapis) Augochlora)). Vicariant events shown in the cladistic analyses are discussed, and an account of distribution is presented.     

Environmental influences on male production and social structure inHalictus rubicundus (Hymenoptera: Halictidae)

Summary Several years' observations of population of a primitively social halictine bee,Halictus rubicundus, revealed the following: (1) there is a correlation between ambient temperatures during the spring provisioning phase and the sex ratio of the resulting brood, such that warmer temperatures are associated with an increase in male bias; (2) over the course of the season, the degree of male bias of eggs laid appears to correspond with photoperiod; and (3) increasing male bias in the first brood is associated with decreasing proportions of social colonies formed in the population, and this effect may be accentuated by small population size. These phenomena suggest that abiotic environmental conditions at the time of brood production may profoundly influence the demography of this population, and that the demography in turn determines the degree to which sociality is expressed. These findings are related to hypotheses dealing with caste determination in temperate halictine bees, viewed in the context of the evolution and maintenance of sociality, and it is proposed that these phenomena reveal a mechanism by which social behavior (the occurrence of a worker caste, in particular) is facultatively fine-tuned to suit the characteristics of the environment.     

A revision of the augochlorine bee genus Chlerogas Vachal (Hymenoptera: Halictidae)

The augochlorine bee genus Chlerogas Vachal is revised and the female is described for the first time. The genus is newly diagnosed and new characters discussed for its separation from other augochlorine genera. Seven new species are recognized– Chlerogas araguaensis, C. boliviensis, C. colombiensis, C. cyaneus, C. ttephos, C. tiara, and C. tozvnesi –in addition to two previously described species: C. chlerogas (Vachal) and C. hirsutipennis Cockerell. The group was previously only recorded from Peru, but is now known in Bolivia, Colombia, Venezuela, and Ecuador. A key is provided for the identification of all species.     

The effect of pollen protein concentration on body size in the sweat bee Lasioglossum zephyrum (Hymenoptera: Apiformes)

Adult bees and wasps provide all the food their offspring require to grow from egg to adult. For a given diet, offspring body size generally increases with an increase in the amount of food consumed as a larva, but the extent to which body size is influenced by the type of food consumed is poorly known. Pollen ranges from 2–60% protein among plant species, and bees are extremely efficient at assimilating nitrogen; therefore, it seems likely that either parent bees adjust the size of larval provisions to compensate for differences in pollen protein concentration or bee offspring attain different body size depending on the pollen type(s) consumed as a larva. We presented the generalist sweat bee Lasioglossum zephyrum with pollen diets that differed in protein content and monitored offspring body size during two experiments. In a protein supplementation experiment, diets ranged from 20–66% protein and consisted of Typha pollen amended with soy protein. On a pollen/soy diet, offspring body size increased 25% with a shift from 20–37% protein, but did not increase further at greater protein concentrations. In a multiple pollen experiment, pollen diets ranged from 20–39% protein and consisted of eight pollens that differed naturally in protein concentration. The largest offspring arose from the most protein-rich pollens, whereas much smaller bees developed on protein-poor pollens. Provision size only predicted offspring size when pollen type, and therefore protein quantity, was considered. Adult foragers did not adjust provision size to compensate for pollen protein. Therefore, offspring body size appears to result from a combination of controlled (provision size) and uncontrolled (pollen quality) factors that arise out of bee foraging decisions.     

Flexible social organization and high incidence of drifting in the sweat bee, Halictus scabiosae

The very diverse social systems of sweat bees make them interesting models to study social evolution. Here we focus on the dispersal behaviour and social organization of Halictus scabiosae , a common yet poorly known species of Europe. By combining field observations and genetic data, we show that females have multiple reproductive strategies, which generates a large diversity in the social structure of nests. A detailed microsatellite analysis of 60 nests revealed that 55% of the nests contained the offspring of a single female, whereas the rest had more complex social structures, with three clear cases of multiple females reproducing in the same nest and frequent occurrence of unrelated individuals. Drifting among nests was surprisingly common, as 16% of the 122 nests in the overall sample and 44% of the nests with complex social structure contained females that had genotypes consistent with being full-sisters of females sampled in other nests of the population. Drifters originated from nests with an above-average productivity and were unrelated to their nestmates, suggesting that drifting might be a strategy to avoid competition among related females. The sex-specific comparison of genetic differentiation indicated that dispersal was male-biased, which would reinforce local resource competition among females. The pattern of genetic differentiation among populations was consistent with a dynamic process of patch colonization and extinction, as expected from the unstable, anthropogenic habitat of this species. Overall, our data show that H. scabiosae varies greatly in dispersal behaviour and social organization. The surprisingly high frequency of drifters echoes recent findings in wasps and bees, calling for further investigation of the adaptive basis of drifting in the social insects.