Standard methods for molecular research in Apis mellifera

From studies of behaviour, chemical communication, genomics and developmental biology, among many others, honey bees have long been a key organism for fundamental breakthroughs in biology. With a genome sequence in hand, and much improved genetic tools, honey bees are now an even more appealing target for answering the major questions of evolutionary biology, population structure, and social organization. At the same time, agricultural incentives to understand how honey bees fall prey to disease, or evade and survive their many pests and pathogens, have pushed for a genetic understanding of individual and social immunity in this species. Below we describe and reference tools for using modern molecular-biology techniques to understand bee behaviour, health, and other aspects of their biology. We focus on DNA and RNA techniques, largely because techniques for assessing bee proteins are covered in detail in Hartfelder et al. (2013). We cover practical needs for bee sampling, transport, and storage, and then discuss a range of current techniques for genetic analysis. We then provide a roadmap for genomic resources and methods for studying bees, followed by specific statistical protocols for population genetics, quantitative genetics, and phylogenetics. Finally, we end with three important tools for predicting gene regulation and function in honey bees: Fluorescence in situ hybridization (FISH), RNA interference (RNAi), and the estimation of chromosomal methylation and its role in epigenetic gene regulation.Fundação para a Ciência e Tecnologi

Langstroth hive construction with cement-vermiculite

Exfoliated vermiculite is a light-weight and cheap product that, because of its thermal resistance, has become a valuable insulating material. With regard to its use in beekeeping, this research tested whether the box for honey bees constructed with cement-vermiculite mortar (CVM) presents physical characteristics similar to those of wood. The experiment was carried out at Seropédica, RJ, Brazil, for eight months. The cement-vermiculite mortar was compared with a control material (pinewood), in the construction of Langstroth boxes and boards, in a completely randomized design, with respect to thermal control, thermal conductivity and its capacity to absorb and lose water. The production cost for a CVM box was estimated. There were no internal temperature differences between CVM and wooden boxes. Thermal conductivity values for CVM and pinewood were similar. CVM absorbed more water and lost water faster than pinewood. Since CVM boxes can be easily constructed, at a low cost and with similar characteristics as traditional boxes, made of wood, the material can be recommended for use in non-migratory beekeeping

The volatile emission of honeybee queens (Apis mellifera L)

The volatile compounds released by honeybee queens (Apis mellifera L) were trapped from the vapour phase with an absorbent (Tenax TA) and extracted in hexane. Heads and tergites of these queens were extracted in dichloromethane. After gas chromatography, the chromatograms were statistically analyzed and compared. Seventy-three percent of the compounds in head extracts and 56% of tergal compounds were found in the trapped volatile signal. There were, however, substantial quantitative differences between the actual amounts of individual compounds found in the trap and in head and tergite extracts respectively. For example, little (E)-9-oxodec-2-enoic acid (9-ODA), the classical queen substance and predominant compound of head extracts, was found in the vapour phase. Tergal signals and mandibular gland secretions contributed equally to the total pheromone blend in the volatile signal. Furthermore, compounds not present in either the tergal or head extracts were found

Genotypic intragroup variance and hoarding behaviour in honeybees (Apis mellifera L.)

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A centromere-specific probe for fluorescence in situ hybridization on chromosomes of Apis mellifera L

Repetitive DNA was isolated through CsCl gradient centrifugation from genomic DNA of Apis mellifera L. DNA fragments were cloned and used for fluorescence in situ hybridization (FISH). Chromosomes were isolated from testes tissue of drone larvae. One clone, GC-6, proved to hybridize to the centromere of 14 chromosomes. This probe could be used in combination with others to identify various chromosomes of Apis mellifera

Mitochondrial DNA length variation in the cytochrome oxidase region of honey bee (Apis mellifera L)

Restriction fragment length polymorphism analysis of Apis mellifera capensis mitochondrial DNA detected a length variation in the cytochrome oxidase I and II gene region. A Bcl I digest revealed a 430-bp insert not shared by A m scutellata and A m carnica

Morphometric analysis of 2 southern African races of honeybee

A suite of 10 morphological characters was identified that discriminated between the 2 southern African honeybee races, Apis mellifera capensis and A m scutellata. Collections of samples from 32 localities which spanned the sub-continent from the west coast to the east coast and ranged from Cape Town in the south to north of Johannesburg were used to define the current distribution of the 2 races and the hybrid zone between them. The data obtained from the morphometric analysis are in good agreement with data on laying worker reproduction

Nestmate Recognition and the Genetic Gestalt in the Mound-Building Ant Formica polyctena

Introduction Social insects live in colonies and with few exceptions they resist mixing with other colonies even in the same species. It has been argued that individuals recognize nest members through a common chemical label or "colony odor". They react with aversion or aggression to conspecifics bearing a foreign label or lacking the appropriate one (Fielde, 1903; Forel, 1923). The recognition labels could be of genetic or environmental origin and a variety of different mechanisms have been found for different species (Wallis, 1962; Mabelis, 1979; Stuart, 1987; Carlin and Hlldobler, 1986). In many cases, transient nestmate recognition cues can be derived from environmental sources such as food or nesting material are capable of overriding other more stable, apparently genetically-based cues, and of inducing aggression among otherwise non aggressive kin (Wilson, 1971; Breed, 1983; Gamboa et al., 1986). Stuart (1987) pointed out that the existence and importance of these cues in a na