Occurrence and morphometry of the hypopharyngeal glands inScaptotrigona postica Lat. (Hymenoptera, Apidae, Meliponinae)

As compared toApis mellifera where only workers have hypopharyngeal glands, inScaptotrigona postica, these glands occur in workers, queens and males. They are composed of two long axial ducts with many unicellular secretory alveoli interconnected by secretory canaliculi. The axial ducts are longer in males than in workers, but the alveolar areas of queens and males are generally smaller. In workers the alveoli have their greatest size in the nurses or middle-aged individuals while in queens and males they are larger in newly emerged individuals. The results indicate that the glands in workers may produce food for the brood as inA. mellifera, since they are well developed in the nurse workers. However, the function of the glands in queens and males remains to be clarified since these individuals have no part in brood care.     

Ultrastructural and cytochemical aspects of metamorphosis in the midgut of Apis mellifera L. (Hymenoptera: Apidae: Apinae)

The midgut of Apis mellifera is remodeled during metamorphosis. The epithelium and, to a lesser extent, the muscular sheath degenerate between the end of the last larval instar and the onset of pupation (prepupa). The larval epithelium is shed to the midgut lumen and digested, while a new epithelium is reconstructed from larval regenerative cells. During pupation, some reorganization still occurs, mainly in brown-eyed pupae. In pharate adult, the midgut wall shows the characteristics of adult, although some cells have pycnotic nuclei. The localization of alkaline and acid phosphatases showed that these enzymes were not involved in the reabsorption of the midgut wall.     

Distribution of acid phosphatases in the hypopharyngeal glands from workers,queens and males of a Brazilian stingless bee Scaptotrigona postica Latreille: an ultrastructural cytochemical study

The present study reports the localization of acid phosphatase in the hypopharyngeal gland cells from workers (newly-emerged, nurse and forager), queens (newly-emerged and laying) and males (newly-emerged and mature for mating) of the Brazilian stingless bee, Scaptotrigona postica. The phosphatase activity varied in intensity and localization depending on the individual class, physiological age and the substrate used. In newly-emerged workers, the phosphatase-positive sites suggest the involvement of the enzyme with cellular differentiation that occurs in the presecretory phase, in nurse workers with protein synthesis and in forager workers with changes in cellular activity or glandular regression. In males mature for mating and laying queens, the positive sites are related to secretory activity, showing that the gland maintains some activity in spite of the regressive aspect. Of the substrates used, beta-glycerophosphate gave the least specific localization.     

The fine structure of the tarsal glands of the honeybee Apis mellifera L. (Hymenoptera)

Summary Tarsal glands are located in the 6th tarsomere of adult honeybee queens, workers and drones. Their structural features are not cast or sex specific. The glandular epithelium is lined by a thin endocuticular layer. A cuticular pocket is formed from a postimaginal delamination of the cuticle secreted by the glandular epithelium. The apical plasma membrane of the glandular cells shows numerous cristae and microvilli lining large crypts that communicate with the subcuticular space. Pinocytotic vesicles, multivesicular bodies and residual dense bodies are present in the apical part of the glandular cells. The RER is well developed in perinuclear and basal parts of the glandular cells, but the Golgi apparatus is a discrete organelle without secretory granules. No exocytotic secretory structures were observed. To reach the glandular pocket, the non-proteinaceous secretory product must pass across the subcuticular space, the cuticular intima, the space between the intima and the cuticular wall, and the cuticular wall of the glandular pocket.     

A comparison of the protein quality of pollens for growth-stimulation of the hypopharyngeal glands and longevity of honey bees,Apis mellifera L. (Hymenoptera: Apidae)

Summary Newly-emerged worker honey bees confined in small cages were fed 33 isonitrogenous diets containing 25 species of pollen. The growth-stimulation of the hypopharyngeal glands was determined by morphological examination after 7 days of feeding. Twenty-two species of pollen were evaluated and compared when they were fed as 10% concentrations of protein, three as 7.5%, six as 5%, two as 2.5%, and one each as 1.25 and 0.625% concentrations.A 0.625% concentration of protein from pollen ofTaraxacum officinale was just as effective in the diet as a 2.5% concentration of protein from pollen ofBaccharis viminea or a 5.0% concentration of protein from pollen ofSorghum. A concentration of 1.25% protein from pollen ofT. officinale in the diet stimulated as much gland growth as either a 7.5% concentration from pollen ofSorghum andB. viminea or a 5.0% concentration of pollen fromZea mays var.saccharata. At a 5.0% concentration of protein, pollen fromRobinia pseudoacacia andPolygonum promoted significantly better development thanRanunculus arvensis andT. officinale fed at the same level. At a 10.0% concentration, pollens fromTrifolium pratense, T. officinale, Prunus serotina, Malus pumila, andPopulus fremonti were significantly better than pollen fromAgoseris, Populus nigra var. italica, orMedicago sativa.Longevity data were obtained for nine species of pollen, six at a concentration of 10% protein, three at 7.5%, four at 5%, and two at 2.5%.

---

Résumé Des ouvrières d'abeilles venant de naître et placées dans des cagettes ont reçu 33 alimentations différentes ayant même teneur en azote et renfermant 25 espèces de pollen. Au bout de 7 jours, la rapidité de croissance des glandes hypopharyngiennes a été étudiée par un examen morphologique. On a comparé la valeur de 22 espèces de pollen administrées aux concentrations en protéine suivantes: 10%, 7,5% pour 3 espèces, 5% pour 6 espèces, 2,5% pour 2 espèces, 1,25 et 0,625% pour 1 espèce chacune.Le pollen deTaraxacum officinale, à une concentration en protéine égale à 0,625%, s'est montré aussi efficace que celui deBaccharis viminea à 2,5% ou celui deSorghum à 5,0%. Le pollen deT. officinale, à une concentration en protéine de 1,25%, a eu le même effet stimulant sur la croissance des glandes que celui deSorghum ouB. viminea à 7,5% ou deZea mays var.saccharata à 5,0%. A une concentration en protéine de 5,0%, les pollens deRobinia pseudoacacia et Polygonum ont donné un meilleur résultat queRanunculus arvensis etT. officinale administrés sous la même forme. A la concentration de 10,0%, les pollens deTrifolium pratense, T. officinale, Prunus serotina, Malus pumila etPopulus fremonti étaient nettement plus efficaces que ceux deAgoseris, Populus nigra var.italica ouMedicago sativa. Des données sur la longévité ont été obtenues pour 9 espèces de pollen, 6 à la concentration en protéine de 10%, 3 à 7%, 4 à 5% et 2 à 2,5%.

     

意大利工蜂咽下腺结构观察

通过研究意大利蜜蜂Apis mellifera工蜂咽下腺的结构及形态,为蜜蜂形态学和蜜蜂生理学以及王浆分泌生理提供新知识采用光学显微镜、扫描电镜、HE染色石蜡切片和透射电镜,从形态学和细胞学水平对意大利工蜂咽下腺的结构和功能做了详尽探究和分析.意大利工蜂咽下腺由多个卵圆形腺泡组成,每个腺泡的直径约为120×180 μm,腺泡之间有微气管相连,腺泡外侧有神经细胞,神经细胞的胞体直径约为12μm.每个腺泡是由多细胞构成,最外层有0.1 ～0.2 μm的结缔组织膜,内部有壁细胞、酶原细胞和王浆分泌细胞,其中王浆分泌细胞分布较多,细胞直径约为16 ～ 18 μm,细胞核直径约为6μm. 由咽下腺的形态结构和细胞类型推断,意大利工蜂咽下腺主要分泌王浆,也具有分泌消化酶的功能,同时了解到分泌消化酶和王浆的途径.     

The vibration dance behavior of queenless workers of the honey bee,Apis mellifera (Hymenoptera: Apidae)

The vibration dance was investigated in queenless (QL) colonies of honey bees. Workers performing the dance had significantly less-developed ovaries than recipients. Vibrators were more likely to be mauled by nestmates (an aggressive act) and were more strongly associated with foraging than were nonvibrating controls. Recipients responded to the dance by increasing the amount of time spent performing tasks. The vibration dance may therefore be associated with aggression in QL colonies and may give workers with less-developed ovaries a degree of control over the behavior of bees with greater ovarian development.     

Ultrastructural modifications in the venom glands of workers of Apis mellifera L. (Hymenoptera: Apidae) promoted by topical application of juvenile hormone

The present study analyzed, the influence of the treatment with juvenile hormone on the ultrastructure of Apis mellifera L. workers' venom glands. Newly emerged workers received topical application of 1 microl of juvenile hormone diluted in hexane, in the concentration of 2 microg/pl. Two controls were used; one control received no treatment (group C1) and other received topical application of 1 microl of hexane (group C2). The aspect of the glandular cells, in not treated newly emerged workers, showed that they are not yet secreting actively. Cellular modifications happened according to the worker age and to the glandular area considered. The most active phase of the gland happened from the emergence to the 14th day. At the 25th day the cells had already lost their secretory characteristic, being the distal area the first to suffer degeneration. The treatment with juvenile hormone and hexane altered the temporal sequence of the glandular cycle, forwarding the secretory cycle and degeneration of the venom gland.     

Transcriptome differences in the hypopharyngeal gland between Western Honeybees (Apis mellifera) and Eastern Honeybees (Apis cerana)

Background

Apis mellifera and Apis cerana are two sibling species of Apidae. Apis cerana is adept at collecting sporadic nectar in mountain and forest region and exhibits stiffer hardiness and acarid resistance as a result of natural selection, whereas Apis mellifera has the advantage of producing royal jelly. To identify differentially expressed genes (DEGs) that affect the development of hypopharyngeal gland (HG) and/or the secretion of royal jelly between these two honeybee species, we performed a digital gene expression (DGE) analysis of the HGs of these two species at three developmental stages (newly emerged worker, nurse and forager).

Results

Twelve DGE-tag libraries were constructed and sequenced using the total RNA extracted from the HGs of newly emerged workers, nurses, and foragers of Apis mellifera and Apis cerana. Finally, a total of 1482 genes in Apis mellifera and 1313 in Apis cerana were found to exhibit an expression difference among the three developmental stages. A total of 1417 DEGs were identified between these two species. Of these, 623, 1072, and 462 genes showed an expression difference at the newly emerged worker, nurse, and forager stages, respectively. The nurse stage exhibited the highest number of DEGs between these two species and most of these were found to be up-regulated in Apis mellifera. These results suggest that the higher yield of royal jelly in Apis mellifera may be due to the higher expression level of these DEGs.

Conclusions

In this study, we investigated the DEGs between the HGs of two sibling honeybee species (Apis mellifera and Apis cerana). Our results indicated that the gene expression difference was associated with the difference in the royal jelly yield between these two species. These results provide an important clue for clarifying the mechanisms underlying hypopharyngeal gland development and the production of royal jelly.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-744) contains supplementary material, which is available to authorized users.     

Ultrastructure and ontogeny of the mandibular glands of the queen honey bee,Apis mellifera L. (Hymenoptera: Apidae)

Each mandibular gland of the queen bee, Apis mellifera L. (Hymenoptera: Apidae) consists of an axial cavity lined with a thin cuticular intima, secreted by a flat epithelium, and numerous glandular units (type 3), each unit with one duct cell and a large polyploid glandular cell. Mitochondria and endoplasmic reticulum are preponderant organelles. During the ontogeny of the queen bee, the ultrastructure of the glandular cells evolves as her level of pheromonal activity. Variations mainly concern the mitochondrial system. Hence, at the beginning of the imaginal life, the homogeneous population of small mitochondria increases. Towards the 3rd–5th day of the queen's imaginal life, the pheromonal activity increases and the mitochondrial differentiation results in the appearance of giant forms. During the highest activity phase (6 to 18-month-old queens), giant mitochondria, associated with endoplasmic reticulum, invade cytoplasmic areas. In queens aged from 18 to 24 months, the reduction of pheromonal and secretory activities is associated with the reduction of mitochondrial population as well as with the accumulation of lipid droplets and various lytic structures.     

Autophagy and apoptosis coordinate physiological cell death in larval salivary glands of Apis mellifera (Hymenoptera: Apidae)

Larval salivary glands of bees provide a good model for the study of hormone-induced programmed cell death in Hymenoptera because they have a well-defined secretory cycle with a peak of secretory activity phase, prior to cocoon spinning, and a degenerative phase, after the cocoon spinning. Our findings demonstrate that there is a relationship between apoptosis and autophagy during physiological cell death in these larval salivary glands, that adds evidence to the hypothesis of overlap in the regulation pathways of both types of programmed cell death. Features of autophagy include cytoplasm vacuolation, acid phosphatase activity, presence of autophagic vacuoles and multi-lamellar structures, as well as a delay in the collapse of many nuclei. Features of apoptosis include bleb formation in the cytoplasm and nuclei, with release of parts of the cytoplasm into the lumen, chromatin compaction, and DNA and nucleolar fragmentation. We propose a model for programmed cell death in larval salivary glands of Apis mellifera where autophagy and apoptosis function cooperatively for a more efficient degeneration of the gland secretory cells.     

Antennal malformations in light ocelli drones of Apis mellifera (Hymenoptera, Apidae)

Malformed antennae of Apis mellifera light ocelli drones were drawn, dissected and mounted permanently on slides containing Canada balsam, in order to count the olfactory discs present in each segment, in comparison with the number of those structures in normal antennae of their brothers. Some drones presented morphological abnormalities in a single segment of the right or left antenna, but others had two or more malformed segments in a same antenna. Drones with malformations in both antennae were also observed. The 4th and 5th flagellum segments were the most frequently affected. In a low number of cases the frequency of olfactory discs in malformed segments did not differ from that one recorded for normal segments. However, in most cases studied, the antennal malformations brought about a significant reduction in the number of olfactory discs from malformed segments.     

A comparative study of morphological integration in Apis mellifera (Insecta, Hymenoptera)

Morphometric correlation matrices from 11 Mediterranean and European honey bee races have been compared. The degree of integration, measured by the eigenvalue variance of the correlation matrix or by Cheverud's index of integration, varies considerably between races but covaries neither with overall body size nor with the variance of the respective sample. Hence, the degree of morphological integration does not depend on body size or the level of variability The patterns of morphometric correlation are significantly similar among all races, but some minor differences in the patterns could be detected: The similarity between the correlation matrices is independent of the degree of phylogenetic relatedness between the respective races. In all races characters belonin to the same functional and/or developmental unit (leg, wing, abdominal characters) have Eiter correlations than the average. Most of the variation in the pattern of correlation can be expfained as a side effect of variation in the degree of integration. Races with high levels of integration tend to have hiher correlations between the main groups of characters, and races with low levels of integration tave smaller correlations between the main grous of characters, while the basic pattern remains undisturbed. In summary, the comparative analysis of morphometric correlation matrices reveals a picture of stability with respect to the pattern of integration and of variation in the degree of integration, which is random with respect to body size, degree of variability, and phylogeny.     

Influence of age and caging upon protein metabolism,hypopharyngeal glands and trophallactic behavior in the honey bee (Apis mellifera L.)

Summary Different-aged honey bees were either kept in a cage together with young sisters for eight days or lived in their colony. Following an injection of¹⁴C-phenylalanine (Phe) we measured incorporation of¹⁴C-Phe into head protein and total protein, as well as the size of the hypopharyngeal glands. While confined in a cage for four hours, injected bees (from colony or cage) dispensed the¹⁴C-labelled protein-rich products of their hypopharyngeal glands to recipients. Eight-day-old colony bees had well developed hypopharyngeal glands, whereas at the age of sixteen days the glands had already decreased in size. Young caged bees had smaller hypopharyngeal glands. Colony bees had higher incorporation rates into total protein and head protein than bees living in a cage. Bees of different age classes, irrespective of caging, fed the same number of recipients; but the amount of¹⁴C-labelled protein-rich jelly distributed by caged bees was significantly smaller than that distributed by colony bees. Our results indicate that trophallaxis between young donor workers and newly emerged recipient worker bees is not the key factor for regular development and activity of the hypopharyngeal glands.Dedicated to Achim Lass's daughter Katrin.     

Juvenile hormone III induced ultrastructural changes in the hypopharyngeal glands of honeybee Apis mellifera L. (Hymenoptera : Apidae) without and with infection by Nosema apis zander (Microsporidae : Nosematidae)

In the fully activated hypopharyngeal gland of the worker honeybee, Apis mellifera (Hymenoptera : Apidae), there were numerous electron-dense secretion granules, large secretion masses, free ribosomes, and arrays of endoplasmic reticulum. Injection of juvenile hormone into the honeybee caused crystallization of the secretion granules. After 1 and 7 days following the injection, both free and attached ribosomes were depleted from the cytoplasm. The depletion of ribosomes from the cytoplasm of the hypopharyngeal gland cells was observed only in honeybees infected by Nosema apis after injection of juvenile hormone. The appearance of an increasing number of lysosome-like bodies in the cytoplasm suggests that juvenile hormone activates the lysozymes, which leads to the degeneration of the hypopharyngeal glands.     

意大利蜜蜂不同日龄工蜂脑部三种miRNA的表达水平

【目的】意大利蜜蜂 Apis mellifera ligustica（简称“意蜂”）是社会性昆虫,蜂群中的工蜂表现出年龄依赖性的行为转变,其行为转变的机制一直是研究的热点。本研究分别检测了3种miRNA（ame-let-7, ame-miR-13b和ame-miR-279）在不同日龄意蜂工蜂脑部的表达情况,以期为探究意蜂工蜂年龄依赖性行为变化机制提供重要线索。【方法】通过荧光定量PCR分别检测了不同发育时期（4, 8, 12, 17, 22, 26和30日龄）意蜂工蜂脑部3种miRNA的表达情况,并检验miRNA的表达差异情况。【结果】ame-let-7的表达量随工蜂日龄的增加逐渐降低,在17日龄后其表达量趋于稳定;ame-miR-13b的表达量则随着工蜂日龄的增加逐渐增加,但在26和30日龄的工蜂中稳定表达;ame-miR-279的表达量则呈类似正态分布状态,其中在12日龄工蜂中表达量最高,且在17日龄后稳定表达。这3种miRNA表达均具有明显的时间特异性。【结论】 ame-let-7, ame-miR-13b和ame-miR-279的表达与工蜂年龄依赖性行为变化具有相关性和规律性,对于深入探究miRNA的作用位点具有重要参考价值。     

Modes of cell death in the hypopharyngeal gland of the honey bee (Apis mellifera l)

Different modes of cell death have been revealed in the regressing hypopharyngeal glands of worker honey bees. The hypopharyngeal gland, which is well developed in young nursing bees to produce protein for larval food, was seen to regress naturally in foraging adult worker bees. A range of techniques including histology, cytochemistry, in situ TUNEL, Annexin V and Comet assays indicated that cells within the gland demonstrate progressive symptoms of apoptosis, necrosis and a vacuolar form of programmed cell death. The latter mode of cell death did not display chromatin margination, but was accompanied by an enhanced level of autophagic and hydrolytic activity in which a cytosolic source of acid phosphatase became manifest in the extra-cisternal spaces. Normal and annexin-positive cells were found to occur in the younger nursing bees, whilst necrosis and an aberrant vacuolar type of apoptosis predominated in the older foraging bees. The relevance of these results to the classification of programmed cell death is discussed.     

中华蜜蜂工蜂咽下腺胚后发育过程中细胞的增殖和凋亡

【目的】咽下腺(hypopharyngeal gland)是蜜蜂重要的外分泌腺,是工蜂合成和分泌蜂王浆的主要腺体。本研究的目的在于了解中华蜜蜂Apis cerana cerana工蜂咽下腺的胚后发育特点。【方法】通过组织形态学、Brd U免疫组织化学和TUNEL细胞凋亡检测等技术,对中华蜜蜂工蜂咽下腺的胚后发育过程及组织结构特点进行了比较研究。【结果】中华蜜蜂工蜂的咽下腺起源自预蛹阶段口器内壁的陷入,细胞分裂活动的高峰期集中在蛹发育的前3 d,随后分裂细胞数减少,并一直持续到蛹发育的第7天左右结束;分泌腺泡的出现大约在蛹发育的第5天。到蛹发育的末期,咽下腺已基本形成,但是没有发育完全;哺育蜂的咽下腺高度发育,分泌活动旺盛;采集蜂的咽下腺中有许多分泌细胞凋亡。【结论】本研究揭示了中蜂工蜂咽下腺胚后发育过程中细胞增殖和凋亡的模式,为昆虫咽下腺的发育和功能研究提供了一定的理论依据。     

Flower visitation patterns of the coexisting honey bees Apis cerana japonica and Apis mellifera (Hymenoptera: Apidae)

We compared flower visitation patterns of two coexisting honey bees, Apis mellifera Linnaeus and Apis cerana japonica Radoszkowski, on 20 plant species, including three exotics, under natural conditions in Nara, Japan, from April to August 2012. We also measured flower color based on bee color vision (15 flower species), nectar volume (nine species) and nectar concentration (eight species). Flowers colored white, pink, red, purple and cream were classified as bee‐blue‐green, and yellow was classified as bee‐green. Apis cerana visited 14 plant species and A. mellifera visited 11. Although the two Apis species are similar in morphology, they visited different plants: in particular, A. cerana visited native plant species more often than did A. mellifera. Both A. mellifera and A. cerana visited not only nectariferous flowers but also those with no nectar. We also found different visitation patterns between A. cerana and A. mellifera: Apis cerana more often visited flowers with smaller color angle (bee‐blue‐green), lower chroma and higher brightness, and flowers secreting nectars of higher concentration and smaller volume than did A. mellifera.