Engrailed expression and body segmentation in the honeybee Apis mellifera

Summary Honeybee embryos were stained with a monoclonal antibody raised against the Drosophila engrailed protein. The antibody was found to label rows of nuclei in the transverse grooves that form the earliest external sign of metameric germ band organization. These grooves demarcate metameric units about seven cell rows wide, of which about three rows with reduced apical cell surfaces account for the grooves. The en stripes appear in the grooves as soon as these form and grow from one to about four cells in width and thus completely overlap the groove. During the rudimentary germ band retraction, the grooves shift slightly backwards relative to both the en stripes and the trachdeal pits. The spatio-temporal pattern by which the series of grooves and stripes arises is quite striking. Both become visible first in the gnathal and thoracic regions, then in the pregnathal parts of the head and in the abdomen. The stripes arise essentially in an antero-posterior sequence. In addition, the earliest stripes to form display a pattern of alternating intensities whereas the later stripes, those in the abdomen, arise with approximately equal strength. The latter trait was earlier observed in the grasshopper, while the former is known from Drosophila where, however, the strong stripes correspond to the weak stripes in the honeybee.     

Local honeybee (Apis mellifera mellifera L.) populations in the Urals

The COI-COII intergenic region of mitochondrial DNA (mtDNA) was studied in local honeybee (Apis mellifera mellifera) L. populations from the Middle and Southern Urals. Analysis of bee colonies in these regions revealed apiaries enriched in families descending from A. m. mellifera in the maternal lineage. These results confirm the suggestion of preservation of A. m. mellifera refuges in the Urals and provide grounds for work on the preservation of the gene pool of this bee variety, valuable for all Russia.     

Changes in food source profitability affect Nasonov gland exposure in honeybee foragers Apis mellifera L.

Summary. When arriving at a known artificial food source, foraging honeybees usually perform circular flights around the feeding place prior to landing. During these flights bees expose their Nasonov gland, an exocrine gland located at the base of the 7^th tergum, that releases a complex blend of volatiles. This behavior may continue even after the bee starts food ingestion. The proportion of bees exposing the Nasonov gland and the duration of its exposure before and during feeding for individual bees were quantified. Trained bees collected sugar solution during 12 visits from a feeder located at 160 m from the hive. Five different reward programs were presented: three constant and two variable. The constant programs offered 0.6, 1.2 or 2.4 M sugar for all 12 visits, while the variable programs delivered either 0.6, 1.2, 0.6 M or 0.6, 2.4, 0.6 M, four visits for each molarity. Results showed that sugar concentration changed the thresholds and durations of Nasonov gland exposure. However, this relationship was found only for Nasonov exposure before bees began to feed. During feeding, a protruded Nasonov gland was only observed for bees that had exposed it prior to feeding; suggesting that Nasonov gland exposure before feeding is a releaser of the during-feeding exposure. In variable reward programs, changes in sugar concentration were followed by changes in both thresholds and durations of exposure. However, Nasonov gland exposure during feeding did not appear to decrease based on measurements of the low profitability during the current foraging visit. These results suggest that Nasonov gland exposure is programmed on the basis of reward expectations, with the bees having acquired this information in the previous foraging visits to the food source.     

Lipophorin of the larval honeybee, Apis mellifera L

Most insects have a major lipoprotein species in the blood (hemolymph) that serves to transport fat from the midgut to the storage depots in fat body cells and from the fat body to peripheral tissues. The generic name lipophorin is used for this lipoprotein. In larvae of the honeybee, Apis mellifera, a lipophorin has been found with properties that correlate well with those of the only other lipophorin reported for an immature insect, that of the tobacco hornworm, Manduca sexta. The honeybee lipophorin (Mr = 530,000) has a density of 1.13 g/ml, contains approximately 41% lipid and 59% protein, and contains two apoproteins, apoLp-I, Mr = 250,000 and apoLp-II, Mr = 80,000, both of which are glycosylated. The lipids consist predominantly of polar lipids, of which phospholipids and diacylglycerols represent 60% of the total. When the intact lipophorin is treated with trypsin, apoLp-I is rapidly proteolyzed, while apoLp-II is resistant, indicating a difference in exposure of the two apoproteins to the aqueous environment. Honeybee apoLp-II cross-reacts with antibodies to M. sexta apoLp-II, but not to anti-M. sexta apoLp-I. No cross-reactivity of honeybee apoLp-I to anti-M. sexta apoLp-I was observed.     

Male fitness of honeybee colonies (Apis mellifera L.)

Honeybees (Apis mellifera L.) have an extreme polyandrous mating system. Worker offspring of 19 naturally mated queens was genotyped with DNA microsatellites, to estimate male reproductive success of 16 drone producing colonies. This allowed for estimating the male mating success on both the colony level and the level of individual drones. The experiment was conducted in a closed population on an isolated island to exclude interferences of drones from unknown colonies. Although all colonies had produced similar numbers of drones, differences among the colonies in male mating success exceeded one order of magnitude. These differences were enhanced by the siring success of individual drones within the offspring of mated queens. The siring success of individual drones was correlated with the mating frequency at the colony level. Thus more successful colonies not only produced drones with a higher chance of mating, but also with a significantly higher proportion of offspring sired than drones from less successful colonies. Although the life cycle of honeybee colonies is very female centred, the male reproductive success appears to be a major driver of natural selection in honeybees.     

Ecdysteroid biosynthesis in workers of the European honeybee Apis mellifera L

We previously reported preferential expression of genes for ecdysteroid signaling in the mushroom bodies of honeybee workers, suggesting a role of ecdysteroid signaling in regulating honeybee behaviors. The organs that produce ecdysteroids in worker honeybees, however, remain unknown. We show here that the expression of neverland and Non-molting glossy/shroud, which are involved in early steps of ecdysteroid synthesis, was enhanced in the ovary, while the expression of CYP306A1 and CYP302A1, which are involved in later steps of ecdysone synthesis, was enhanced in the brain, and the expression of CYP314A1, which is involved in converting ecdysone into active 20-hydroxyecdysone (20E), was enhanced in the brain, fat body, and ovary. In in vitro organ culture, a significant amount of ecdysteroids was detected in the culture medium of the brain, fat body, and hypopharyngeal glands. The ecdysteroids detected in the culture medium of the fat body were identified as ecdysone and 20E. These findings suggest that, in worker honeybees, cholesterol is converted into intermediate ecdysteroids in the ovary, whereas ecdysone is synthesized and secreted mainly by the brain and converted into 20E in the brain and fat body.     

引入西方蜜蜂对中蜂的危害及生态影响

作者阐述自1896年中国引进西方蜜蜂Apis melliferaL.的优良品种如意大利蜂Apis mellifera ligusticaSpinola和喀尼阿兰蜂Apis mellifera Carnica Pollmann以来,使当地的东方蜜蜂Apis cerana F.受到严重危害,其分布区域缩小75%以上,种群数量减少80%以上。使山林植物授粉总量减少,导致植物多样性减少。文中提出: 建立自然保护区保存本地蜜蜂遗传特性,和采用基因转移等技术,培育具有西方蜜蜂优良生产性能的中蜂新品种,逐步恢复中蜂的种群数量。     

Stratification and synaptogenesis in the mushroom body of the honeybee,Apis mellifera

Stratification is a basic anatomical feature of central brain in both vertebrates and many invertebrates. The aim of this study was to investigate the relationship between stratification and synaptogenesis in the developing mushroom bodies of the honeybee. During metamorphosis, the vertical lobe of mushroom body shows progressive stratification with three thick primary strata and more secondary strata and laminae. Three primary strata are formed at the metamorphic stage P1, before the youngest generation of the mushroom body intrinsic neurons, Kenyon cells, is produced. Thus, the primary strata within the lobe are unlikely to represent three major subpopulations of the Kenyon cells sequentially produced in the mushroom bodies. Formation of laminae starts at the stage P2 and culminates at the end of metamorphosis. The laminae appear within the lobe rather than being added sequentially from the ingrowth stratum. Alternating dark and light lamina (lamina doublets) are formed in the vertical lobe in late metamorphosis (stages P6–P9), but they are not visible in adults. The pattern of stratification is not continuous along the vertical lobe at the same developmental stage, and resorting of axons of the Kenyon cells is likely to occur within dark laminae. In the developing vertical lobe, dark laminae show lower synaptic density and exhibit an ultra structure that is indicative for a delay in synaptogenesis relative to the primary strata. A local transient block of synaptogenesis within the dark laminae may provide correct targeting of Kenyon cells by extrinsic mushroom body neurons. J. Morphol., 2010. © 2010 Wiley‐Liss, Inc.     

Genetic dissection of honeybee (Apis mellifera L.) foraging behavior

We demonstrate the effects of a new quantitative trait locus (QTL), designated pln3, that was mapped in a backcross population derived from strains of bees selected for the amount of pollen they store in combs. We independently confirmed pln3 by demonstrating its effects on individual foraging behavior, as we did previously for QTLs pln1 and pln2 (Hunt et al. 1995). QTL pln2 is very robust in its effects on foraging behavior. In this study, pln2 was again shown to affect individual foraging behavior of workers derived from a hybrid backcross of the selected strains. In addition, pln2 was shown to affect the amount of pollen stored in combs of colonies derived from a wide cross of European and Africanized honeybees. This is noteworthy because it demonstrates that we can map QTLs for behavior in interstrain crosses derived from selective breeding and study their effects in unselected, natural populations. The results we present also demonstrate the repeatability of finding QTLs with measurable effects, even after outcrossing selected strains, suggesting that there is a relatively small subset of QTLs with major effects segregating in the population from which we selected our founding breeding populations. The different QTLs, pln1, pln2, and pln3, appear to have different effects, revealing the complex genetic architecture of honeybee foraging behavior.     

Behavioural discrimination of oilseed rape volatiles by the honeybee Apis mellifera L.

This study investigated the ability of the honeybee to discriminatebetween six compounds previously identified as oilseed rapefloral volatiles: linalool, 2-phenylethanol, methyl salicylate,benzyl alcohol, (E)-2-hexenal and 1-octen-3-ol. These componentswere tested individually or in a synthetic mixture for theirability to elicit the conditioned proboscis extension response.Three experiments were done: conditioning to the mixture atone concentration (1.0, 0.1 or 0.01 µg per component)and testing to the individual components either at the conditioningor at the other concentrations; conditioning to individual componentsand testing to the mixture (1.0 µg); conditioning andtesting to the individual components (1.0 µg). The resultsfrom the proboscis extension assay were then compared to thoseobtained from free-flying bees in a flight room. From the conditionedproboscis extension assay, a conditioning threshold level wasfound (0.1 µg for the mixture studied) below which a reliableconditioning could not be achieved. Recognition thresholds wereobserved: bees responded to concentrations ten-fold higher orlower than that used for conditioning. Responses to lower concentrationswere weaker, whilst responses were increased at concentrationshigher than the conditioning one. A hierarchy within the compoundstested was found, with linalool, 2-phenylethanol and methylsalicylate cueing mixture recognition more effectively thanthe other components. The ranking order of the six componentswas similar in both the conditioned proboscis extension andthe conditioned foraging behaviour in flight room. When conditionedand tested to the individual components, bees discriminateda learned odour from a number of others. However, the specificitylevel for the recognition of the learned odour varied accordingto the component, the most clearly discriminated being the compoundswhich were used by the bee in mixture recognition.     

Differential parallel processing of olfactory information in the honeybee, Apis mellifera L.

Two distinct neuronal pathways connect the first olfactory neuropil, the antennal lobe, with higher integration areas, such as the mushroom bodies, via antennal lobe projection neurons. Intracellular recordings were used to address the question whether neuroanatomical features affect odor-coding properties. We found that neurons in the median antennocerebral tract code odors by latency differences or specific inhibitory phases in combination with excitatory phases, have a more specific activity profile for different odors and convey the information with a delay. The neurons of the lateral antennocerebral tract code odors by spike rate differences, have a broader activity profile for different odors, and convey the information quickly. Thus, rather preliminary information about the olfactory stimulus first reaches the mushroom bodies and the lateral horn via neurons of the lateral antennocerebral tract and subsequently odor information becomes more specified by activities of neurons of the median antennocerebral tract. We conclude that this neuroanatomical feature is not related to the distinction between different odors, but rather reflects a dual coding of the same odor stimuli by two different neuronal strategies focusing different properties of the same stimulus.     

A microsatellite-based linkage map of the honeybee, Apis mellifera L

A linkage map for the honeybee (Apis mellifera) was constructed mainly from the progeny of two hybrid queens (A. m. ligustica x A. m. mellifera). A total of 541 loci were mapped; 474 were microsatellite loci; a few were additional bands produced during PCRs, one of the two rDNA loci (using ITS), the MDH locus, and three sex-linked markers (Q and FB loci and one RAPD band). Twenty-four linkage groups were estimated of which 5 were minute (between 7.1 and 22.8 cM) and 19 were major groups (>76.5 cM). The number of major linkage groups exceeded by three the number of chromosomes of the complement (n = 16). The sum of the lengths of all linkage groups amounts to 4061 cM to which must be added at least 320 cM to link groups in excess, making a total of at least 4381 cM. The length of the largest linkage group I was 630 cM. The average density of markers was 7.5 cM and the average resolution was about one marker every 300 kb. For most of the large groups, the centromeric region was determined genetically, as described in (accompanying article in this issue), using half-tetrad analysis of thelytokous parthenogens in which diploid restoration occurs through central fusion. Several cases of segregation distortion that appreared to result from deleterious recessives were discovered. A low positive interference was also detected.     

茚虫威对意大利蜜蜂毒性研究

通过单次饲喂高浓度茚虫威药液以及连续饲喂亚致死浓度茚虫威药液进行染毒,记录和分析急性慢性经口毒性试验中,意大利蜜蜂的中毒症状和死亡情况;测定慢性毒性试验中意大利蜜蜂每天药液消耗量,并计算危害商值进行初步风险评价。结果表明,98%茚虫威原药对意大利蜜蜂的半致死剂量48 h-LD_(50)为0.399μg a.i./bee,每日半数致死剂量240 h-LDD_(50)为0.0233μg a.i./bee/day,其中慢性毒性试验中1.66 mg a.i./L处理组的意大利蜜蜂死亡率在144 h之后呈现显著性的增加,240 h达到70%,且该处理组意大利蜜蜂的摄食量为17.6μL,少于对照组和其他处理组。此外,茚虫威根据目前推荐使用剂量计算出的危害商值基本大于50。茚虫威对意大利蜜蜂急性毒性(48 h)是高毒,存在较高的风险;亚致死浓度在144 h后会对意大利蜜蜂造成较大的威胁。     

The mating behaviour of the honeybee (Apis mellifera L.)

An account is given of the mating responses of free-flying drones towards a queen, and of the pheromones controlling these responses.     

Genetic determination of honeybee (Apis mellifera L.) foraging preferences

Progeny from different honeybee queens that were reared in, and foraged from, the same colony sometimes differed in their floral preferences, confirming that these are to some extent innately determined.     

Associative learning and discrimination of motion cues in the harnessed honeybee Apis mellifera L.

We previously studied a conditioning paradigm to associate the proboscis extension reflex (PER) with monochromatic light (conditioned stimulus; CS) in harnessed honeybees. Here, we established a novel conditioning paradigm to associate the PER with a motion cue generated using graphics interchange format (GIF) animations with a speed of 12 mm/s speed and a frame rate of 25 Hz as the CS, which were projected onto a screen consisting of a translucent circular cone that largely covered the visual field of the harnessed bee using two liquid crystal projectors. The acquisition rate reached a plateau at approximately 40% after seven trials, indicating that the bees were successfully conditioned with the motion cue. We demonstrated four properties of the conditioning paradigm. First, the acquisition rate was enhanced by antennae deprivation, suggesting that sensory input from the antennae interferes with the visual associative learning. Second, bees conditioned with a backward-direction motion cue did not respond to the forward-direction, suggesting that bees can discriminate the two directions in this paradigm. Third, the bees can retain memory for motion cue direction for 48 h. Finally, the acquisition rate did not differ significantly between foragers and nurse bees. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Interadult feeding of jelly in honeybee (Apis mellifera L.) colonies

Summary Honeybee nurses (8 days old) were injected with ¹⁴C-phenylalanine. These bees then dispensed the ¹⁴C-labelled protein-rich products of their hypopharyngeal glands to the queen and the brood, and also to young drones and workers of all age classes. In small colonies containing 400–800 bees, nearly one-quarter of the radioactivity which could not be recovered in the nurses was fed by them in a protein-bound form to other members of the worker caste. During one night, one nurse fed an average of 4–5 foragers with proteinaceous food. The role of nurses in the work allotment system of honeybee colonies therefore needs a new, extended definition. Nurses are largely responsible for preparing nutrients from pollen, which is difficult to digest. They then distribute the nutritionally valuable protein produced by their hypopharyngeal glands to practically all hive mates.Dedicated to Professor Dr. O. Kepka on the occasion of his 65th birthday