CO2麻醉对中华蜜蜂处女蜂王卵巢发育的影响

【目的】明确中华蜜蜂Apis cerana cerana蜂王羽化后生殖系统的动态变化,探索CO₂麻醉对中华蜜蜂处女蜂王卵巢发育的影响。【方法】通过人工育王技术和蜂王的储存获取不同日龄的中华蜜蜂处女蜂王,观察蜂王羽化后生殖系统的形态,测定生殖系统各项形态学指标(体重、卵巢重、受精囊直径和卵巢管数目)。中华蜜蜂处女蜂王连续2 d(分别于5和6日龄时)接受不同浓度(50%, 75%与90%)CO₂麻醉8 min, 以及不同日龄处女蜂王接受75% CO₂麻醉1~3次(每次8 min),待蜂王发育至10日龄时,利用石蜡切片技术与HE染色法观察蜂王卵巢形态、测定蜂王卵巢重和蜂王卵巢最大卵母细胞长度,通过实时荧光定量PCR(RT-qPCR)测定10日龄蜂王头部卵黄原蛋白基因(Vg)的相对表达量。【结果】中华蜜蜂蜂王的卵巢管数目与受精囊直径不会随着日龄发生明显的波动,但蜂王卵巢重在6-12日龄明显增加,之后将趋于稳定。与对照组(未接受CO₂麻醉处理)相比,50%, 75%与90% CO₂麻醉后中华蜜蜂蜂王卵巢重与最大卵母细胞长度均增加,并且头部Vg基因的相对表达量上调。其中CO₂浓度为75%,麻醉次数2次显著促进了中华蜜蜂蜂王的卵巢发育。【结论】中华蜜蜂蜂王羽化后卵巢会发生明显的发育变化,而CO2麻醉能加速卵巢的发育。     

How to fool a virgin: Artificial dealation triggers oviposition in virgin Lasius niger queens

Once inseminated, ant queens rapidly shed their wings and start to lay eggs. Here we test whether there is a causal link between dealation and oviposition in the ant Lasius niger. We show that artificially dealated virgin queens start to lay eggs shortly after wing removal, whereas winged virgin queens hardly ever lay eggs. Dealate virgins do, however, produce fewer eggs than mated queens. These findings indicate that dealation does induce egg-laying, and that other factors, such as mating and/or insemination, further stimulate oviposition under natural conditions. Received 2 January 2006; revised 8 February 2006; accepted 4 April 2006.     

授粉用明亮熊蜂的人工饲养技术

为提高授粉用熊蜂人工繁育效率,降低生产成本,对明亮熊蜂Bombus lucorumL.人工饲养中诱导蜂王产卵和人工控制下的交配2个技术环节进行了研究。结果表明,采用诱导器和伴饲2~3只蜜蜂工蜂的诱导方法,蜂王产卵率和蜂群成群率最高,成群时间最短;使用塑料诱导器和木制诱导器的效果没有显著差异。将8~9日龄的蜂王和11~12日龄的雄蜂放入交配笼时,其交配成功率最高。交配笼内的蜂王数量应控制在30只/m3左右。在晴天,交配笼放置在室外(758 000 lux),其交配成功率最高;在阴天,交配笼应放在交配室内,并开启荧光灯照明(35 000 lux),其交配成功率最高。     

Increasing the availability of food and the reproduction ofPoecilus cupreus in a cereal field by strip-management

The spruce bud moth,Zeiraphera canadensis Mutt & Free. (Lepidoptera: Tortricidae), displayed a bimodal activity pattern in two young white spruce,Picea glauca (Moench) Voss, plantations. During the day, when temperatures were highest and relative humidities lowest, most moths remained in the lower crown, presumably in response to the risk of hygrothermal stress and/or predation. Flight activity peaks occurred near dawn and dusk. Moths moved higher in the crowns shortly after sunset and many were observed flying above the tree canopy. This activity peak was associated with oviposition and mating. Oviposition started at sunset and continued for approximately 3 h. Females probed oviposition sites with their ovipositor before ovipositing, suggesting that they use sensillae on the ovipositor to evaluate potential oviposition sites. Most mating occurred between 22∶00 and 04∶00 h. Males hovered adjacent to branches containing females before alighting. After alighting, males usually approached to within 2–5 cm of females and then remained motionless for a short time before attempting mounting, indicating that some short-range communication usually occurs prior to mating. Reduced flight activity between midnight and dawn was not due to low temperatures, which were above the minimal temperatures for flight for males (10°C) and females (12°C) determined in the laboratory.     

Differential circular RNAs expression in ovary during oviposition in honey bees

Circular RNAs (circRNAs) are non-coding RNAs newly identified and play important roles in RNA regulation. The mechanism and function of circRNAs have been reported in some species. However, little is known regarding circRNAs in honey bees. In this study, we analyzed circRNAs through bioinformatics, and predicted 12,211 circRNAs in the ovary of honey bee queens. 1340, 175 and 100 circRNAs were differentially expressed in comparisons of egg-laying queens vs virgin queens, egg-laying inhibited queens vs egg-laying queens and egg-laying recovery queens vs egg-laying inhibited queens. Further, functional annotation of differentially expressed circRNAs revealed several pathways that are closely related to ovary activation and oviposition, including insulin secretion and calcium signaling pathways. Moreover, the potential interactions among circRNAs, miRNAs, lncRNAs and mRNAs were investigated. Ame_circ_0005197 and ame_circ_0016640 were observed to sponge several reproductive related miRNAs. These findings demonstrate that circRNAs have potential effects in ovary activation and oviposition of honey bees.     

Queen power in relation to age and mating status in the Argentine antIridomyrmex humilis (Mayr)

Summary Queen power inIridomyrmex humilis during six arbitrarily chosen physiological stages of queens (virgin winged queens at the time of emergence, the same 4–5 days old, mated winged queens 5–6 days old, mated queens at the time of dealation, young egg-laying queens, and old egg-laying queens) was tested with regard to workers. Tested workers and queens were nestmates. The results were as follows. 1) Power of the old queens remained rather constant throughout the reproductive season. 2) Young queens were always less attractive than older ones. No changes were observed from the time of emergence to dealation. 3) At the time of egg-laying, these young queens became markedly attractive for workers but never as much as old egg-laying queens. Therefore, insemination and dealation do not trigger the increase in queen power. In contrast, oogenesis ending by egg-laying produces a significant increase in queen power.     

Selection for non-diapause in the bumblebee Bombus terrestris, with notes on the effect of inbreeding

Bumblebees are generally believed to be annual insects. However, here we will show that under laboratory conditions the bumblebee Bombus terrestris (L.) can produce a second generation without a period of cold storage (diapause) or CO₂ narcosis (a method to break diapause). It is also shown that this so-called non-diapause trait can be selected for. The percentage of non-diapausing queens increased from 8% (minimum) to 97% (maximum) in two generations of selection. However, it was not possible to maintain isofemale non-diapause lines. Colonies of the fourth and fifth generation remained small (expressed in worker number) and produced a small number of queens. Also the percentage of queens that started laying eggs (defined as the percentage non-diapause) decreased in the fourth and fifth generations. To study whether this decline of the non-diapause lines was caused by inbreeding, a control experiment was conducted. In this control experiment queens were mated with their brothers (full-sib mating) for several generations and the number of queens that start egg laying was measured. This revealed that inbreeding can have a negative effect on the egg-laying capacities of queens thus causing the decline of inbred (non-diapause) lines.     

The effects of temperature on development of the large narcissus fly (Merodon equestris)

Eggs, larvae, pupae and adults of the large narcissus fly (Merodon equestris) were reared at a series of constant temperatures between 9–24°C. Egg development required from 37 days at 9°C to 7 days at 21.5°C. The low-temperature threshold for development was 6.7°C. Larvae reared at 1424°C were fully-grown after 18 weeks, but it took much longer for such insects to pupate, and adult flies emerged only after about 45 weeks of development. Large narcissus flies enter diapause during the larval stage and overwinter as fully-fed larvae, forming pupae in the following spring. Post-winter pupation and pupal development took from 169 days at 10°C to 36 days at 21.5°C. Of this, pupal development required from 91 days at 10°C to 19 days at 21.5°C. The low-temperature threshold for post-winter pupation and pupal development was 7.1°C, and for pupal development alone, 7.2°C. Females maintained at or below 19°C laid few eggs, whereas some females kept at or above 21.5°C laid more than 100 eggs (mean 69 ± 36). Approximately 50% of females maintained at or above 21.5°C laid less than 10 eggs during their lifetime. The mean egg-laying time was 6 to 9 days. Although temperatures at or below 19°C inhibited mating, once a female had mated, such temperatures did not prevent oviposition.     

Comparing Alternative Methods for Holding Virgin Honey Bee Queens for One Week in Mailing Cages before Mating

In beekeeping, queen honey bees are often temporarily kept alive in cages. We determined the survival of newly-emerged virgin honey bee queens every day for seven days in an experiment that simultaneously investigated three factors: queen cage type (wooden three-hole or plastic), attendant workers (present or absent) and food type (sugar candy, honey, or both). Ten queens were tested in each of the 12 combinations. Queens were reared using standard beekeeping methods (Doolittle/grafting) and emerged from their cells into vials held in an incubator at 34C. All 12 combinations gave high survival (90 or 100%) for three days but only one method (wooden cage, with attendants, honey) gave 100% survival to day seven. Factors affecting queen survival were analysed. Across all combinations, attendant bees significantly increased survival (18% vs. 53%, p<0.001). In addition, there was an interaction between food type and cage type (p<0.001) with the honey and plastic cage combination giving reduced survival. An additional group of queens was reared and held for seven days using the best method, and then directly introduced using smoke into queenless nucleus colonies that had been dequeened five days previously. Acceptance was high (80%, 8/10) showing that this combination is also suitable for preparing queens for introduction into colonies. Having a simple method for keeping newly-emerged virgin queens alive in cages for one week and acceptable for introduction into queenless colonies will be useful in honey bee breeding. In particular, it facilitates the screening of many queens for genetic or phenotypic characteristics when only a small proportion meets the desired criteria. These can then be introduced into queenless hives for natural mating or insemination, both of which take place when queens are one week old.     

Physiological regulation of long-term oviposition in the house cricket, Acheta domesticus

In house crickets [Acheta domesticus (L.)] a single mating early in adult life sufficed to induce egg laying for the duration of the life of a female. Female house crickets mated readily shortly after adult emergence but oviposition did not commence until about 12–14 days after emergence, even though females matured eggs by 7 days. The egg-laying factor associated with mating remained active during prolonged periods of substrate deprivation during which the female did not oviposit. If the spermatophore was removed prematurely shortly after a mating, the long-term, egg-laying response was truncated and was correlated with a dramatic decline in the fertility of eggs which were oviposited. The egg-laying stimulus appeared to act in the spermatheca, apparently through neural means, since denervation of the spermatheca abolished mating-induced oviposition. These results indicate that the oviposition factor found in the testes is able to act for long periods of time and has to be present continually in order to be effective. Furthermore, the long-term oviposition stimulus in the house cricket may be different from prostaglandin E2 which induces a prompt ovipositional response.     

Oviposition in Triatoma protracta: Role of mating and relationship to egg growth

Yolk deposition begins in the terminal oocytes of virgins of Triatoma protracta a few days after adult eclosion, and while a few eggs may be matured before vitellogenesis ceases, none are laid. Mating at day four stimulates egg maturation and oviposition in fed and unfed females, egg-laying beginning as early as nine days after eclosion. If mating is delayed until day 16, by which time vitellogenesis normally has ceased, stored eggs are laid within two to four days and yolk deposition is resumed. Removal of the brain prevents oviposition, as does the severance of the ventral nerve cord. Thus, an intact central nervous system is required for egg-laying. Also, since neither operation inhibits egg maturation or ovulation, it appears that the latter reproductive responses to mating are independent of oviposition.     

Influence de modifications des conditions lumineuses sur les rythmes circadiens de vitellogenese et d'ovulation chez Drosophila melanogaster

Under photoperiodic conditions (LD 12:12), a rhythm was observed in the frequencies of ovarian egg chambers and of mature oöcytes. Females reared and kept in permanent darkness (DD) did not show any rhythm. After a transition from LD 12:12 to DD, the rhythm of vitellogenesis remained almost unchanged for at least 5 days while the rhythm of oöcytes retention disappeared.Suppression of a suitable oviposition substrate resulted in an accumulation of mature oöcytes in the ovaries. When a conveneint medium was given again, the egg-laying proved to be highly dependent on the light conditions. Most of the oöcytes remained in retention during the light phase. Significant egg-laying only occurred after the beginning of darkness. In such conditions females can lay one egg every 3 min.The egg-laying rhythm observed under cyclic light conditions thus arises from two separate physiological processes: oöcyte production (vitellogenesis) which has a circadian, endogenous rhythm and oviposition which is directly dependent on the light conditions.     

兰州熊蜂蜂王头部产卵相关microRNA生物信息学分析

【目的】头部是熊蜂感觉和取食中心,也是生理行为的指挥中心。microRNA（miRNA）参与重要生理行为的调控。本研究初步探索了产卵和未产卵兰州熊蜂Bombus lantschouensis蜂王头部差异表达miRNA及其靶基因,以期探明蜂王头部miRNA在产卵过程中作用。【方法】以本土优良蜂种兰州熊蜂 B. lantschouensis 为材料,利用Solexa高通量测序技术对产卵和未产卵蜂王头部miRNA进行测序,运用生物信息学软件对miRNA及其靶基因进行预测;并应用qPCR技术验证极显著差异表达的miRNA。【结果】共获得兰州熊蜂产卵和未产卵蜂王头部miRNA的clean data数,分别为14 228 864和21 431 031 条reads;长度分析表明,在19~24 nt序列中22 nt序列数量最多,分别占产卵和未产卵蜂王序列的45.8%和45.1%。miRNA预测结果显示,共获得297个miRNA,其中270个为已知miRNA,有92个存在于蜜蜂中,其余178个分别存在于蜜蜂以外的其他物种中;另外27个为新的miRNA。在蜜蜂92个已知的miRNA中,在产卵蜂王头部中表达的共有91个,特异表达的有2个;在未产卵蜂王头部中表达的共有90个,特异表达的有1个。27个新的miRNA中,在产卵蜂王头部中表达的共有22个,特异表达的有2个;未产卵蜂王头部中表达的共有25个,特异表达的有5个。miRNA差异表达分析表明,在产卵与未产卵蜂王头部,共有8个已知miRNA表达量达到差异极显著水平（P<0.01）。qPCR结果表明,这8个表达差异极显著的miRNA均存在于熊蜂体内。在8个差异极显著的miRNA中,3个miRNA进行靶基因预测,发现它们共同调控9个靶基因,这些基因主要参与GTP结合和转录调控。【结论】本研究首次利用高通量测序技术鉴定了熊蜂头部组织的miRNA,并发现蜂王产卵与否受到miRNA差异表达的调控。结果为今后深入开展熊蜂miRNA功能验证及产卵调控机制研究奠定了基础。     

Biological activity of five strains ofMetarhizium anisopliae,upon the coffee berry borerHypothenemus hampei (Col.: Scolytidae)

Five strains of the entomopathogenic fungusMetarhizium anisopliae (Metschnikoff) Sorokin were bioassayed against the coffee berry borerHypothenemus hampei (Ferrari) (Coleoptera: Scolytidae) under laboratory conditions (T=27±2°C; R.H.=90±5% and 12∶12 L∶D light cycle). The LC₅₀ values for the three most virulent strains, Ma4, Ma5 and Ma3, were 4.2, 5.9 and 6.7×10⁶ conidia/ml of suspension respectively and median lethal times (LT₅₀) observed were between 9.7 and 13.8 days. The maximum percentage sporulation was obtained with strains Ma3 and Ma4 both with 90% sporulation and Ma10 with 60.7%. These results suggest that the fungusM. anisopliae could be incorporated into an integrated control programme for the biological control ofH. hampei.

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Résumé Cinq souches du champignon entomopathogèneMetarhizium anisopliae (Metschnikoff) Sorokin, ont été évaluées dans les conditions du laboratoire, pour mesurer leur activité vis-à-vis du scolyte des fruits du caféierHypothenemus hampei (Ferrari) (Coleoptera: Scolytidae). Les résultats montrent que les souches Ma4, Ma5 et Ma3 sont les plus virulentes contreH. hampei, avec des valeurs de concentration létale moyenne (CL₅₀, valeurs minimales et maximales, ± limite de confiance à 95%) de 0,0226 (0,0195–0,00260), 0,0328 (0,0267–0,0390) et 0,0354 (0,0297–0,0417) équivalentes à 4,2, 5,9 et 6,7×10⁶ conidies/ml de suspension et un temps létal moyen (TL₅₀) entre 9,7 et 13,8 jours. Les pourcentages maximaux de sporulation ont été obtenus avec les souches Ma3 et Ma4 avec 90%, la souche Ma10 présentant le pourcentage le plus bas de sporulation (67%). Ces résultats démontrent queM. anisopliae peut être introduit dans un programme de lutte intégrée contreH. hampei.

     

Biology ofMicroplitis mediator [Hym.: Braconidae] a gregarious endoparasite ofAgrotis segetum [Lep.: Noctuidae]

Microplitis mediator (Haliday) a gregarious endoparasite was recorded for the first time fromAgrotis segetum (Schiff) in Ankara, Turkey. The female parasites found their hosts by responding to the faeces of the caterpillars. An average, females laid 15.5±1.6 eggs in the bodies of their hosts. The newly laid eggs were elongated, oval in shape and 0.23±0.004 mm long and 0.07 mm wide. They hatched in 5, 4 and 3 days at 20±2°C, 25±2°C and 30±2°C respectively when maintained at 60–70% R.H. and 14∶10 light∶dark regime. At the same temperatures, the larval stage lasted for 24.9±0.6, 18.2±0.4 and 17.1±0.5 days respectively. The prepupal stage was completed in 2 days at 25±2°C, whereas the prepupal and pupal (cocoon) stage lasted 10.9±0.2, 7.0±0.1 and 6.2±0.1 days respectively at the temperatures mentioned above. The adults started mating and feeding shortly after emergence. Female parasites started laying after one day, 7–11 hours and 5–7 hours at the temperatures stated above. At these temperatures females lived for 10.8±0.2, 5.4±0.1, 4.6±0.1 days and laid on average 556, 484 and 363 eggs respectively, whereas the males survived 10.5±0.3, 4.7±0.1 and 4.4±0.1 days respectively.      

Adult response of the almond seed wasp,Eurytoma amygdali, to chemicals from its host and certain nonhosts

Females of the almond seed wasp,Eurytoma amygdali Enderlein (Hymenoptera, Eurytomidae), responded in an olfactometer positively to odours from almond flowers and unripe fruits, but not to almond leaf odours and odours from flowers and unripe fruits of certain other nonhostPrunus species. Males responded to none of these odours. Extracts of undamaged unripe almond fruits (using ethanol, methanol, acetone, hexane, dichloromethane, or petroleum ether) stimulated female aggregation on glass surfaces treated with these extracts; in addition, certain fruit extracts (ethanol, methanol, or acetone) stimulated oviposition. Extracts of undamaged almond leaves (ethanol, methanol, or acetone) and flowers (ethanol or methanol) also stimulated female aggregation and oviposition. Aggregation and oviposition in response to an ethanol extract of almond fruits was intense in females aged 5 to 14 days and from 12∶00 to 18∶00h (photophase between 06∶00 and 20∶00). Certain almond fruit (ethanol, methanol, acetone or hexane) and flower extracts (ethanol or methanol) also provoked female response in the olfactometer. The results strongly suggest that certain chemical stimuli emanating from parts of the almond tree play a major role in host selection and oviposition. Some of the extracts tested may be a good source for the isolation, identification and synthesis of compounds stimulating attraction, aggregation and oviposition in nature. *** DIRECT SUPPORT *** A3414024 00003     

Biological studies including population growth parameters of the predatory miteAmblyseius barkeri [Acarina.: Phytoseiidae] at 25°C in the laboratory

Amblyseius barkeri (Hughes) fed onThrips tabaci (Lind.) at 25°C showed an average duration of 2.2, 0.8 and 3.2 days for the egg, larval and nymphal stages, with mortalities at 1.0, 1.0 and 3.1%, respectively. Females represented 63% of the population and required multiple matings for optimal fertility. The oviposition period was 20.3 days and the average oviposition rate 2.3 eggs per day. The intrinsic rate of increase was 0.22 per day. The expected life span was 29.6 days for ♀♀ and 27.4 days for ♂♂.A. barkeri ♂♂ and ♀♀ both consumed 3.3 nymphs of thrips per day (mean value for the feeding stages). The larva does not take up food. In the absence of thripsA. barkeri was able to consume two-spotted spider mites,Tetranychus urticae (Koch), and their eggs, adult broad mites,Polyphagotarsonemus latus (Banks), and pollen of various plants. Cannibalism was observed when food was lacking. Certain morphological features, egglaying, mating and predatory behaviour are described.      

Social control of egg-laying rate in queens of the fire ant, Solenopsis invicta*

ABSTRACT Social control of egg-laying rate in queens of the fire ant (Solenopsis invicta Buren) was studied by experimental manipulation of the number of larvae, pupae and workers in colonies, and the age and size of larvae and workers. Workers and pupae do not stimulate oviposition by queens. The number of fourth instar larvae, on the other hand, bears a positive log-log relationship to the queen's egg-laying rate. Such larvae are needed both to stimulate and maintain oviposition. Their withdrawal results, within 48 h, in a decline in queen oviposition almost to zero. Their addition to broodless nests results in peak laying in about 4 days. Larvae in the first three stadia and early in the fourth stadium have a much lower effect upon queen fecundity. Sexual larvae are only c. 5% as stimulating on a weight basis, but equivalent on an individual basis. Several associated measures are positively correlated to egg-laying rate: weight of the queen, the number of her vitellogenic follicles per ovariole, total vitellogenic follicles, the time she spends feeding and (usually) the number of workers in the retinue that cares for her. The egg volume is negatively correlated with laying rate, so that queens lay more eggs for the same expenditure of material as laying rate increases. As body size of workers increases, they become less effective in transmitting the larval stimulation to the queen, but worker age has no effect on this ability. For a given number of larvae, queens in small, naturally growing colonies lay fewer, larger eggs than do queens in experimental colonies, but their fecundity increases more rapidly in relation to number of larvae. When larvae are fed vital-dyed food in one experimental colony, and then transferred to an undyed colony, the dye is rapidly transferred to worker crops, and into the queen's eggs, indicating bulk movement of material from larvae to workers to the queen and eggs. Large larvae are more effective at this than small larvae. Fourth instar larvae may be a digestive and metabolic caste that processes protein for egg production by the queen. If that is the case, the queen and fourth instar larvae are linked in a positive feedback loop. Either the logarithmic relation of fecundity to larval numbers or physical limits of the queen may set the maximum egg-laying rate, and thus determine maximum colony size. The data do not allow a clear choice between these alternatives.     

Queen fecundity and reproductive skew in the termite Nasutitermes corniger

A major goal of studies on social animals is to understand variation in reproduction within and between groups. We used hierarchical regressions to analyze oviposition rates in the neotropical termite Nasutitermes corniger, a species with both monogynous and polygynous colonies. Queen fecundity was a non-linear, saturating function of queen weight. Greater queen weight was associated with larger nest size and with lower numbers of queens per nest, suggesting competition among queens for resources acquired by the colony. The collective egglaying rate of pairs of queens exceeded that of single queens, but further increases in queen number did not raise total fecundity. Skew in oviposition rates, as quantified by Morisita’s index I _δ, averaged 1.2, indicating inequalities in reproductive rates that are only moderately greater than expected for random apportionment. The leveling off of oviposition with increasing queen weight suggests that it is costly for individual females to produce eggs at high rates, which could favor tolerance of reproduction by other females, reducing reproductive skew. We hypothesize that the incentive to tolerate reproduction by other females is especially pronounced for heavier queens, because these queens are close to the limit of their own reproductive capacity. Consistent with this hypothesis, skew in oviposition rates was inversely related to the mean weight of queens within a nest. Received 8 March 2007; revised 17 September 2007; accepted 3 October 2007.     

熊蜂气管内寄生螨布赫纳蝗螨的生物学特性观察

对新疆与河北地区野生明亮熊蜂及小峰熊蜂蜂王内寄生螨布赫纳蝗螨Locustacarus buchneri的生物学特性进行了初步观察。结果表明： 该螨对两种熊蜂的寄生率分别为10.43%和8.3%,寄生部位为气管与气囊。室内饲养至30天的蜂王体内寄生螨量明显高于饲养至60天的蜂王。未被螨寄生的蜂王与被寄生者比较,前者体重显著高于后者,但二者间产卵始期、产卵率、食卵率和存活率差异不显著。