Functional genomics in the wild: a case study with paper wasps shows challenges and prospects for RNA interference in ecological systems.

RNA interference (RNAi) is a useful tool to assess gene function by knocking down expression of a target gene and has been used successfully in domestic and laboratory organisms. However, the use of RNAi for functional genomics has not fully extended into ecological model organisms in natural environments. Assessment of gene function in the wild is important because gene function can be environmentally and context dependent. Here, we present a case study using RNAi to assess gene function in wild paper wasps Polistes metricus, to test roles for two candidate genes (NADH dehydrogenase (NADHdh) and retinoid and fatty acid binding protein (RfaBp)) in the development of reproductive castes. Previous studies have shown that these genes are upregulated in larvae that become queens compared to workers, but this pattern was reversed in the laboratory, making field-based studies necessary. We orally administered dsRNA to larvae in field colonies and found evidence of a short-term knockdown followed by a compensatory rebound in expression for RfaBp. We also observed the predicted worker-like decrease in lipid stores in NADHdh dsRNA treated wasps, suggesting a possible role for NADHdh in caste development. We discuss our results in the context of challenges for using RNAi for functional genomics in ecological model organisms in the field.

Candidate genes for individual recognition in Polistes fuscatus paper wasps.

Few animals are known to individually recognize conspecifics, i.e. learn and recall unique individuals during subsequent encounters, and nearly all are social vertebrates. Remarkably, the social paper wasp Polistes fuscatus has recently been discovered to possess this ability, which is useful for remembering identities during competitive social interactions. We analyzed brain gene expression in staged encounters between pairs of individuals to explore potential mechanisms underlying wasps' ability to recall familiar individuals using real-time qRT-PCR. We identified four candidate genes (IP3K, IP3R, Nckx30C and Su(var)2-10) that were down-regulated in the presence of familiar individuals compared to single wasps and pairs of wasps meeting for the first time. These candidate genes are related to calcium signaling, therefore, we treated wasps with lithium chloride, a pharmacological agent that inhibits calcium signaling in neurons. This treatment decreased aggression in paper wasps, but did not affect expression of genes related to calcium signaling. The results suggest calcium signaling differences may be related to individual memory recall in wasps, and we present four promising candidate genes for future study. These data suggest genes associated with dominance behavior may be co-opted for individual recognition, but further work is needed to establish a causal association with the behavior.

Formation of nanoparticles by ion beam irradiation of thin films.

The possibility of fabricating nanoparticles by ion bombardment was investigated by the ion bombardment of indium films on oxide covered Si and Cr surfaces. The different masses of implanting specimen ensured the different energy transfer while the same Si substrate ensured the same thermal conductivity for the In and Cr layers. Chromium served as a reference for the effect of ion bombardment and as a substrate as well. The SRIM program was used to simulate the ion surface interaction process. The nanoparticles were detected by scanning electron microscopy (SEM). We found that the melting of the In layer results in the formation of nanoparticles of 50-300 nm diameter and 5-10 nm height. This method can be promising for nanoparticle formation of materials with low melting point.

Nutrition and division of labor: Effects on foraging and brain gene expression in the paper wasp Polistes metricus.

Deeply conserved molecular mechanisms regulate food-searching behaviour in response to nutritional cues in a wide variety of vertebrates and invertebrates. Studies of the highly eusocial honey bee have shown that nutritional physiology and some conserved nutrient signalling pathways, especially the insulin pathway, also regulate the division of labour between foraging and non-foraging individuals. Typically, lean workers leave the nest to forage for food, and well-nourished workers perform tasks inside the nest. Here we provide the first direct test of whether similar mechanisms operate in a primitively eusocial insect in an independently evolved social lineage, the paper wasp Polistes metricus. We found that food deprivation caused reduced lipid stores and higher levels of colony and individual foraging. Individuals with greatly reduced lipid stores foraged at extremely elevated levels. In addition, brain expression of several foraging-related genes was influenced by food deprivation, including insulin-like peptide 2 (ilp2). Together with previous findings, our results demonstrate that nutrition regulates foraging division of labour in two independently evolved social insect lineages (bees and wasps), despite large differences in social organization. Our results also provide additional support for the idea that nutritional asymmetries among individuals, based on differences in nutritional physiology and expression of conserved nutrient signalling genes in the brain, are important in the division of labour in eusocial societies.

Evaluating Native Bee Communities and Nutrition in Managed Grasslands.

Native pollinators are important for providing vital services in agroecosystems; however, their numbers are declining globally. Bees are the most efficient and diverse members of the pollinator community; therefore, it is imperative that management strategies be implemented that positively affect bee community composition and health. Here, we test responses of the bee and flowering plant communities to land management treatments in the context of grasslands in the upper Midwestern United States, a critical area with respect to bee declines. Twelve sites were selected to examine floral resources and wild bee communities based on three different types of grasslands: tallgrass prairie remnants, ungrazed restorations, and grazed restorations. Total bee abundance was significantly higher in ungrazed restorations than remnants, but there were no significant differences among grasslands in community composition or Shannon diversity. Across the three grassland types we also examined mass and lipid stores as nutritional health indicators in three sweat bees (Halictidae), Augochlora pura, Agapostemon virescens, and Halictus ligatus. Although there were no differences in lipid content, total average bee mass was significantly higher in Ag. virescens collected from ungrazed restorations as compared to remnants. Floral abundance of native and non-native species combined was significantly higher in grazed restorations compared to remnants and ungrazed restorations. However, ungrazed restorations had higher abundance and richness of native flowering ramets. These data suggest that bee abundance and nutrition are driven by high abundance of native flowering plant species, rather than total flowering plants.

Ion mixing at 20 keV: a comparison of the effects of Ga+, Ar+ and CF4+ ion irradiation.

Medium-energy (some tens of keV) ion irradiation is frequently used in various technologies. It is well known that during this irradiation serious alterations are introduced to the material, changing its structure, composition, etc. While there are studies on the amorphization, no results have been reported on the medium-energy ion beam-induced mixing, however. In this work, we present Auger electron spectroscopy (AES) depth profiling measurements of Si/Cr multilayer samples, which were irradiated by various ions (Ga+, Ar+, CF4+) of 20 keV applying angles of incidence of 5 degrees (Ga+), 65 degrees (Ga+) and 75 degrees (Ar+, CF4+). The ion beam-induced mixing at the Si/Cr interface (the broadening of the interface) was measured as a function of the removed layer thickness. The weakest and strongest ion mixing (for a given removed layer thickness) were found for CF4+ and Ga+ 5 degrees irradiations, respectively. In the case of Ga+ irradiation, the larger the angle of incidence the weaker the ion mixing. The extent of mixing does not correlate with the corresponding projected range. Comparison of the experimentally measured ion mixed profiles with those given by dynamic TRIM simulations gave poor agreement for Ar+ and fails for Ga+ irradiations, respectively.

Formation of ge nanocrystals in SiO2 by electron beam evaporation.

Ge nanocrystals were formed by electron beam evaporation on SiO2 covered Si substrates. The size and distribution of the nanocrystals were studied by atomic force microscopy, scanning electron microscopy and cross-sectional transmission electron microscopy. Dependencies of the nanocrystal size, of the nanocrystal surface coverage, and sheet resistance obtained by van der Pauw method of the Ge layer have been found on the evaporation time. The suggested growth mechanism for the formation of nanocrystals is the Volmer-Weber type. The sheet resistance exhibited a power dependence on the nanocrystal size.

Evolution of magnetism on a curved nano-surface.

To design custom magnetic nanostructures, it is indispensable to acquire precise knowledge about the systems in the nanoscale range where the magnetism forms. In this paper we present the effect of a curved surface on the evolution of magnetism in ultrathin iron films. Nominally 70 Å thick iron films were deposited in 9 steps on 3 different types of templates: (a) a monolayer of silica spheres with 25 nm diameter, (b) a monolayer of silica spheres with 400 nm diameter and (c) for comparison a flat silicon substrate. In situ iron evaporation took place in an ultrahigh vacuum chamber using the molecular beam epitaxy technique. After the evaporation steps, time differential nuclear forward scattering spectra, grazing incidence small angle X-ray scattering images and X-ray reflectivity curves were recorded. In order to reconstruct and visualize the magnetic moment configuration in the iron cap formed on top of the silica spheres, micromagnetic simulations were performed for all iron thicknesses. We found a great influence of the template topography on the onset of magnetism and on the developed magnetic nanostructure. We observed an individual magnetic behaviour for the 400 nm spheres which was modelled by vortex formation and a collective magnetic structure for the 25 nm spheres where magnetic domains spread over several particles. Depth selective nuclear forward scattering measurements showed that the formation of magnetism begins at the top region of the 400 nm spheres in contrast to the 25 nm particles where the magnetism first appears in the region where the spheres are in contact with each other.

Simulation of the backscattered electron intensity of multi layer structure for the explanation of secondary electron contrast.

The intensities of the secondary electrons (SE) and of the backscattered electrons (BSE) at energy 100 eV have been measured on a Ni/C/Ni/C/Ni/C/(Si substrate) multilayer structure by exciting it with primary electrons of 5, 2.5 and 1.25 keV energies. It has been found that both intensities similarly vary while thinning the specimen. The difference as small as 4 nm in the underlying layer thicknesses resulted in visible intensity change. Utilizing this intensity change, the thickness difference of neighboring regions could be revealed from the SE image. No simple phenomenological model was found to interpret the change of intensity, thus the intensity of the BSE electrons has been calculated by means of a newly developed Monte Carlo simulation. This code also considers the secondary electron generation and transport through the solid. The calculated and measured intensities agree well supporting the validity of the model.

Evo-devo and the evolution of social behavior: brain gene expression analyses in social insects.

Studies of genes and social behavior, aided by new genomic resources, are coming of age. Here, we show how some of the insights that have emerged from research on the evolution of development (evo-devo) also provide a useful framework for studying the evolution of social behavior at the molecular level. These insights include co-opting old genes for new functions, phenotypic modularity, genetic tool kits, the importance of gene regulation in evolutionary change, and the influences of some genes over multiple timescales. We next outline a few differences between development and behavior that pose challenges for an evo-devo approach to behavior. For the remainder of this chapter, we review several studies that illustrate the relevance of evo-devo insights to our understanding of the evolution of behaviors related to eusociality in the insect societies.

Highly Uniform Epitaxial ZnO Nanorod Arrays for Nanopiezotronics.

Highly uniform and c-axis-aligned ZnO nanorod arrays were fabricated in predefined patterns by a low temperature homoepitaxial aqueous chemical method. The nucleation seed patterns were realized in polymer and in metal thin films, resulting in, all-ZnO and bottom-contacted structures, respectively. Both of them show excellent geometrical uniformity: the cross-sectional uniformity according to the scanning electron micrographs across the array is lower than 2%. The diameter of the hexagonal prism-shaped nanorods can be set in the range of 90-170 nm while their typical length achievable is 0.5-2.3 mum. The effect of the surface polarity was also examined, however, no significant difference was found between the arrays grown on Zn-terminated and on O-terminated face of the ZnO single crystal. The transmission electron microscopy observation revealed the single crystalline nature of the nanorods. The current-voltage characteristics taken on an individual nanorod contacted by a Au-coated atomic force microscope tip reflected Schottky-type behavior. The geometrical uniformity, the designable pattern, and the electrical properties make the presented nanorod arrays ideal candidates to be used in ZnO-based DC nanogenerator and in next-generation integrated piezoelectric nano-electromechanical systems (NEMS).

Carbohydrate metabolism genes and pathways in insects: insights from the honey bee genome.

Carbohydrate-metabolizing enzymes may have particularly interesting roles in the honey bee, Apis mellifera, because this social insect has an extremely carbohydrate-rich diet, and nutrition plays important roles in caste determination and socially mediated behavioural plasticity. We annotated a total of 174 genes encoding carbohydrate-metabolizing enzymes and 28 genes encoding lipid-metabolizing enzymes, based on orthology to their counterparts in the fly, Drosophila melanogaster, and the mosquito, Anopheles gambiae. We found that the number of genes for carbohydrate metabolism appears to be more evolutionarily labile than for lipid metabolism. In particular, we identified striking changes in gene number or genomic organization for genes encoding glycolytic enzymes, cellulase, glucose oxidase and glucose dehydrogenases, glucose-methanol-choline (GMC) oxidoreductases, fucosyltransferases, and lysozymes.