-Within the western honey bee (Apis mellifera ), there are more than 20 recognised subspecies. It is well known that these subspecies differ in their wing venation patterns. However, there is a demand for efficient tools to identify honey bee subspecies, ecotypes, populations or hybrids. The aim of this study was to develop a fast and easy identification method based on analysing forewing vein patterns of honey bees by geometric morphometrics. Reference samples for the subspecies were obtained from the Morphometric Bee Data Bank in Oberursel, Germany. These contained 187 honey bee colonies allocated into 25 subspecies from four evolutionary lineages. The identification of evolutionary lineages of honey bees based on forewing venations proved to be highly reliable, which confirms earlier studies. The accuracy of honey bee subspecies identification was less consistent and ranged from 100 to 50% and was particularly low in African honey bees. The obtained identification data were exported to the IdentiFly computer software, which is freely available.honey bee / subspecies discrimination / geometric morphometry / wing morphometry

Regulatory RNAs exert their cellular functions through RNA-binding proteins (RBPs). Identifying RNA-protein interactions is therefore key for a molecular understanding of regulatory RNAs. To date, RNA-bound proteins have been identified primarily through RNA purification followed by mass spectrometry. Here, we develop incPRINT (in cell protein-RNA interaction), a high-throughput method to identify in-cell RNA-protein interactions revealed by quantifiable luminescence. Applying incPRINT to long noncoding RNAs (lncRNAs), we identify RBPs specifically interacting with the lncRNA Firre and three functionally distinct regions of the lncRNA Xist. incPRINT confirms previously known lncRNA-protein interactions and identifies additional interactions that had evaded detection with other approaches. Importantly, the majority of the incPRINT-defined interactions are specific to individual functional regions of the large Xist transcript. Thus, we present an RNA-centric method that enables reliable identification of RNA-region-specific RBPs and is applicable to any RNA of interest.

Small noncoding RNAs perform multiple regulatory functions in cells, and their exogenous mimics are widely used in research and experimental therapies to interfere with target gene expression. MicroRNAs (miRNAs) are the most thoroughly investigated representatives of the small RNA family, which includes short interfering RNAs (siRNAs), PIWI-associated RNA (piRNAs), and others. Numerous methods have been adopted for the detection and characterization of small RNAs, which is challenging due to their short length and low level of expression. These include molecular biology methods such as real-time RT-PCR, northern blotting, hybridization to microarrays, cloning and sequencing, as well as single cell miRNA detection by microscopy with in situ hybridization (ISH). In this review, we focus on the ISH method, including its fluorescent version (FISH), and we present recent methodological advances that facilitated its successful adaptation for small RNA detection. We discuss relevant technical aspects as well as the advantages and limitations of ISH. We also refer to numerous applications of small RNA ISH in basic research and molecular diagnostics.

Wild bees in natural conditions can develop under various environmental stressors. Heavy metal pollution of the environment is one of the most widely studied stressors in insects, yet its effect is poorly described in bees. We have measured how pollution of the environment along a zinc, cadmium and lead contamination gradient in Poland affects bee development, using red mason bees (Osmia bicornis) as a model and their forewing asymmetry measures to assess possible developmental instabilities. We have also described wing asymmetry measures in the red mason bee—an important managed pollinator species—for the first time. The development of bee larvae in a contaminated environment did not affect forewing asymmetry measures, but it did lead to a negative correlation of wing size with contamination in females. Bees also showed a clear change in their asymmetry measures between various seasons, suggesting other, unknown environmental factors affecting wing asymmetry more than pollution. Sexes were found to have different forewing shape and size, larger females having larger forewings than the smaller males. The direction of size asymmetry was in favour of the left side in both sexes and also shape differences between the left and right wings showed similar tendencies in males and females. The levels of forewing shape and size asymmetry were smaller in females, making them the more symmetrical sex.

Biological invasions provide a unique opportunity to gain insight into basic biological processes occurring under new circumstances. During the process of establishment, exotic species are exposed to various stressors which may affect their development. Presence of the stressors is often detected by measurements of left-right body asymmetry, which consists of two main components: fluctuating asymmetry and directional asymmetry. Fluctuating asymmetry constitutes random differences between the two body sides, whereas directional asymmetry occurs when a particular trait is bigger on one of the sides. The relation between these two asymmetry components is still not fully understood. Our goal was to investigate the potential differences in asymmetry patterns between native and invasive populations of Tetropium fuscum (Fabr. 1787) (Coleoptera: Cerambycidae), a harmful forest pest native to Europe and introduced to North America. Wing asymmetry assessment was based on the geometric morphometrics of hind wings. We found that specimens from invaded area were markedly smaller and have more asymmetric wings than individuals from native population, suggesting some unfavorable conditions in the invaded area. Moreover, we found significant directional asymmetry in the native but not in the invasive population. On the other hand, differences between left and right hind wings were similar in the native and invasive populations, in terms of direction. This suggests that a high level of fluctuating asymmetry in the invasive population may blur the intrinsic directional asymmetry and hinder its detection. Our data show that fluctuating asymmetry has a potential as an indicator of developmental stress in invasive species.