蜂王受精囊维持精子可育性的生理基础研究进展

蜜蜂是社会性群居昆虫,蜂群中通常只有1头生殖系统发育健全、具有繁殖能力的雌性个体——蜂王,保持蜂王旺盛的产卵力是蜂群繁衍和发展的重要保障.受精囊是蜂王交配后存储精子的结构,它可以维持精子可育性长达数年之久,其生理结构和功能值得深入探讨.本文综述了近年来蜜蜂受精囊结构与生殖生理、精子活力的影响因素和蜂王受精囊微环境的生理功能等领域的研究进展,旨在为蜜蜂种质资源评价、保护和利用以及害虫防治策略提供指导.     

利用VNTR分子标记鉴定蜜蜂群内蜂王交配次数和雄蜂母系来源

如何准确测定蜂王交配次数和雄蜂母系来源,是研究蜜蜂亚家系行为生物学的关键。本研究利用王浆主蛋白（MRJPs）的串联重复序列多态性（VNTR）分子标记分别鉴定了蜂王单雄人工授精、双雄人工授精和自然交尾的中华蜜蜂Apis cerana cerana蜂群中的蜂王交配次数和雄蜂母系来源。结果表明： 在蜂王单雄人工授精和双雄人工授精蜂群中,蜂王的交配次数分别为1和2;在蜂王自然交尾的2个蜂群中,蜂王的交配次数分别为8和5。另外,经鉴定发现：在以上实验蜂群中,所有雄蜂都是由蜂王产的未受精卵发育而来。因此,作为一种分子标记,蜜蜂MRJPs VNTR能简单、有效地鉴定蜂群内蜂王的交配次数和雄蜂母系来源。     

蜜蜂的交配行为

蜜蜂是一妻多夫制，处女蜂王可与７－１７只雄蜂交配。说细介绍了雄蜂内阳茎的构造，交配时内阳茎翻出体外的过程，蜂王和雄蜂交配的情况，一次交配后留在蜂王蜇针腔的交配标志，以后与蜂王交配的雄蜂如何将堵塞在蜇针腔的交配标志除去的过程。     

蜂群中蜜蜂个体之间的关系

蜜蜂是营群体生活的昆虫。一个蜜蜂群体实际上是一个家庭,由一个母亲(蜂王)、几个父亲(雄蜂、交配后死亡)和它的后代(工蜂和雄蜂)组成。初看起来它们的关系很简单,但由于蜂王的交配方式不同,蜂王可以产受精卵和未受精卵,而且蜜蜂还存在着单倍、二倍性,蜜蜂家庭中蜜蜂个体的关系却相当复杂。认识一个蜂群中可能存在的关系,有助于了解蜂群的遗传组成,对于蜜蜂遗传育种有重要意义。近几十年来,经过各国蜜蜂遗传育种学家的不断努力,才逐渐搞清楚蜂群中蜜蜂个体的关系。为了说明这个问题,这里简单介绍一下蜜蜂的细胞学、蜜蜂卵子和精子的发生以及蜂王的交配方式。蜜蜂的细胞学对大多数动物来说,一个新生命是由一个精细胞和一个卵细胞结合而来。每种性细胞各含有一组染色体,因而给予新的个体两组染色体,称为二倍体数。蜂王和工蜂是雌性蜜蜂,它     

蜜蜂繁殖冲突与雌性蜜蜂信息素研究进展

在营造社会性生活的蜜蜂群体里,蜂王释放出蜂王信息素来控制工蜂卵巢发育及改造王台特性,并吸引雄蜂为之交配,使蜂群正常繁衍。本文在国内外相关研究的基础上,对蜜蜂各蜂种、亚种以及特殊蜂群无政府主义蜂群、海角蜜蜂等雌性蜜蜂信息素成分以及含量变化进行综合论述,并对蜂王主要信息素对工蜂和雄蜂生理影响以及今后的研究趋势作扼要介绍。     

婚飞行为影响中华蜜蜂性成熟处女蜂王的基因表达

婚飞是性成熟处女蜂王与雄蜂交配过程中的一个重要前奏, 在该过程中蜂王体内伴随着一系列重要的生理变化。为了探究中华蜜蜂Apis cerana cerana处女蜂王婚飞过程中基因表达变化, 本研究利用数字基因表达谱(digital gene expression, DGE) 技术分析了中华蜜蜂性成熟处女蜂王飞行与未飞行之间的基因表达差异。经DGE测序, 分别从两个样品中获得5.98和6.01 百万条Clean标签。通过分析检测到250个基因有差异表达, 其中133个基因在飞行蜂王中上调表达, 117个基因在飞行蜂王中下调表达。这些差异基因可以归类到348个功能性类别和142个生化途径。结果表明中华蜜蜂性成熟处女蜂王在婚飞过程中大量基因的表达发生了变化。这些结果为进一步研究中华蜜蜂蜂王婚飞过程中生理变化的分子机制提供了重要的基因表达信息。     

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。     

中华蜜蜂交配和产卵行为生态学研究

研究了中华蜜蜂(Apis cerana cerana Fab.)蜂王与雄蜂交配行为生态以及蜂王产卵行为生态。结果表明,温度对中华蜜蜂蜂王和雄蜂封盖子期影响,导致蜂王初生重、性成熟时间差异显著(P<0．05),蜂王和雄蜂认巢飞行次数分别为1．23～1．31和1．08～1．13,持续时间分别为0．12～0．13和0．16～0．20h;蜂王和雄蜂交配飞行次数分别为1．10～1．12和1．01～1．05,持续时间分别为0．22～0．23和0．18～0．23h;蜂王与雄蜂交配最适宜温度为20～28℃,蜂王交配飞行一次侧输卵管的精子数为3．37×10^6～4．15×10^6,自然交配产卵蜂王受精囊精子数为3．55×10^6～3．62×10^6蜂王初生重与产卵量之间呈正相关,周年蜂王产卵量受气候和蜜粉源影响明显。     

翼手目动物的特殊生殖策略：精子贮存

介绍了精子贮存现象在翼手目动物中的分布、产生的原因、精子贮存的部位、贮存后的受精能力、精子长期贮存的机制及贮存过程中的免疫问题,并对将来的研究方向进行了展望。     

中华蜜蜂与意大利蜜蜂蜂王体表信息素含量比较

【目的】分析中华蜜蜂Apis cerana cerana与意大利蜜蜂Apis mellifera ligustica蜂王不同时期的体表信息素含量变化,探索两蜂种之间蜂王交尾干扰机理。【方法】本试验通过GC-MS检测并分析了中华蜜蜂蜂王和意大利蜜蜂蜂王刚出房、性成熟时期以及婚飞过程中体表信息素含量变化。【结果】研究结果表明:中华蜜蜂性成熟蜂王在飞行过程中,其体表9-ODA含量显著高于刚出房蜂王,9-HDA显著高于刚出房蜂王和性成熟蜂王;意大利蜜蜂飞行蜂王在9-ODA含量也显著高于刚出房蜂王。另外,意大利蜜蜂性成熟期蜂王在9-ODA、9-HDA、10-HDA含量显著高于中华蜜蜂蜂王,而两蜂种蜂王体表信息素在婚飞时期差异不显著。【结论】同种蜂王不同发育时期,其体表信息素含量存在差异;中华蜜蜂蜂王与意大利蜂王在婚飞过程中,其体表信息素差异不显著,但部分体表信息素在性成熟而未进行婚飞时差异显著。     

Multiple paternity in a natural population of a salamander with long-term sperm storage

Sperm competition appears to be an important aspect of any mating system in which individual female organisms mate with multiple males and store sperm. Post-copulatory sexual selection may be particularly important in species that store sperm throughout long breeding seasons, because the lengthy storage period may permit extensive interactions among rival sperm. Few studies have addressed the potential for sperm competition in species exhibiting prolonged sperm storage. We used microsatellite markers to examine offspring paternity in field-collected clutches of the Ocoee salamander (Desmognathus ocoee), a species in which female organisms store sperm for up to 9 months prior to fertilization. We found that 96% of clutches were sired by multiple males, but that the majority of females used sperm from only two or three males to fertilize their eggs. The high rate of multiple mating by females suggests that sperm competition is an important aspect of this mating system. Comparison of our data with those of other parentage studies in salamanders and newts reveals that multiple mating may be common in urodele amphibians. Nevertheless, the number of males siring offspring per clutch in D. ocoee did not differ appreciably from that in other species of urodeles with shorter storage periods.     

Stored sperm differs from ejaculated sperm by proteome alterations associated with energy metabolism in the honeybee Apis mellifera

Sperm are exposed to substantially different environments during their life history, such as seminal fluid or the female sexual tract, but remarkably little information is currently available about whether and how much sperm composition and function alters in these different environments. Here, we used the honeybee Apis mellifera and quantified differences in the abundance and activity of sperm proteins sampled either from ejaculates or from the female’s sperm storage organ. We find that stored and ejaculated sperm contain the same set of proteins but that the abundance of specific proteins differed substantially between ejaculated and stored sperm. Most proteins with a significant change in abundance are related to sperm energy metabolism. Enzymatic assays performed for a subset of these proteins indicate that specific protein activities differ between stored and ejaculated sperm and are typically higher in ejaculated compared to stored sperm. We provide evidence that the cellular machinery of sperm is plastic and differs between sperm within the ejaculate and within the female’s storage organ. Future work will be required to test whether these changes are a consequence of active adaptation or sperm senescence and whether they alter sperm performance indifferent chemical environments or impact on the cost of sperm storage by the female.However, these changes can be expected to influence sperm performance and therefore determine sperm viability or sperm competitiveness for storage or egg fertilization.     

Seminal proteins but not sperm induce morphological changes in the Drosophila melanogaster female reproductive tract during sperm storage

In most insects, sperm transferred by the male to the female during mating are stored within the female reproductive tract for subsequent use in fertilization. In Drosophila melanogaster, male accessory gland proteins (Acps) within the seminal fluid are required for efficient accumulation of sperm in the female's sperm storage organs. To determine the events within the female reproductive tract that occur during sperm storage, and the role that Acps and sperm play in these events, we identified morphological changes that take place during sperm storage in females mated to wild-type, Acp-deficient or sperm-deficient males. A reproducible set of morphological changes occurs in a wild-type mating. These were categorized into 10 stereotypic stages. Sperm are not needed for progression through these stages in females, but receipt of Acps is essential for progression beyond the first few stages of morphological change. Furthermore, females that received small quantities of Acps reached slightly later stages than females that received no Acps. Our results suggest that timely morphological changes in the female reproductive tract, possibly muscular in nature, may be needed for successful sperm storage, and that Acps from the male are needed in order for these changes to occur.     

Sperm storage by females of the polyandrous noctuid moth Heliothis virescens

Female tobacco budworm moths, Heliothis virescens, generally mate with more than one male, receiving from each mate both fertilizing sperm (eupyrene) and nonfertilizing anucleate sperm (apyrene), which is thought to play a role in sperm competition. One male typically gains sperm precedence, but it is not consistently the last or the first male to mate. I investigated the mechanism of this variable pattern of paternity by examining the patterns of storage of both types of sperm in the female's spermatheca as a function of multiple mating and male phenotype. The number of stored apyrene sperm varied with mating history, being greatest in twice-mated females and least in females mated to one nonvirgin male. In contrast, only one ejaculate's worth of eupyrene sperm was stored regardless of female mating history (once or twice mated). Thus, while they store two complements of apyrene sperm, twice-mated females apparently store only one ejaculate's worth of eupyrene sperm. This biased pattern of sperm storage may contribute to the variable pattern of paternity observed in this species. Eupyrene sperm storage also correlated positively with female size, male age and spermatophore size. Finally, a new sperm storage site was identified and described. It is a bulged region in the seminal duct. Copyright 2000 The Association for the Study of Animal Behaviour.     

Reproductive biology and function of multiple mating in the mating system of a tree cricket,Truljalia hibinonis (Orthoptera: Podoscritinae)

A tree cricket,Truljalia hibinonis, is known to show a novel sperm removal during copulation. The pattern of copulations and ovipositions showed that the sperm removal functioned to increase reproductive success for sperm removing males. The sperm removal by males evolves under the system in which female accept multiple mating. The possible benefits of multiple mating for females are examined. Multiple mating did not seem to be necessary for avoiding sperm depletion, because females stored huge number of sperm in their sperm storage organ after finishing oviposition. The ingestion of metanotal secretion during copulation also had no effect on increasing fecundity and egg size. However, mating experience may have a positive effect on increasing fecundity slightly, though there were no differences between once- and twice-mated females. The other possible benefits for each male and female are discussed.     

Sperm storage and utilization in female Queensland fruit flies (Bactrocera tryoni)

Abstract.  Female animals that use sperm from a single mating to fertilize eggs over an extended period require efficient mechanisms for sperm storage and use. There have been few studies of these mechanisms in tephritid flies. Mating, copula duration, sperm storage and sperm usage patterns are assessed in an Australian tephritid, the Queensland fruit fly ( Bactrocera tryoni ; a.k.a. 'Q-fly'). In particular, the present study investigates whether each of these aspects of mating varies in relation to female size or male size, whether sperm storage patterns change over time after mating (1, 5, 10 and 15 days), and the relative roles of the ventral receptacle and the two spermathecae as sperm storage organs. Large females are more likely to mate than are small females, and are also more fecund in the first 5 days after mating. Females are more likely to store some sperm and, among those that store some sperm, store more sperm if their mate is large. Most sperm are stored in the spermathecae (median = 97%), often with high levels of asymmetry between the two spermathecae. Asymmetry of sperm storage is related to number of sperm stored, but not to male or female size. Total number of stored sperm declines over the 15 days after mating, but this decrease in sperm numbers only reflects changes in the spermathecae; numbers of sperm in the ventral receptacle remain unchanged over this period. As a consequence, the proportion of total sperm stored in the spermathecae declines relative to the ventral receptacle. These results are consistent with a system in which small numbers of sperm are maintained in the ventral receptacle for fertilizations, and are replenished by sperm from the spermathecae as required. Sperm distribution and usage patterns in Q-flies are comparable with recent findings in medflies, Ceratitis capitata , but differ markedly from patterns found in several Anastrepha species.     

Factors Affecting Sperm Quality Before and After Mating of Calopterygid Damselflies

Damselflies (Odonata: Zygoptera) have a more complex sperm transfer system than other internally ejaculating insects. Males translocate sperm from the internal reproductive organs to the specific sperm vesicles, a small cavity on the body surface, and then transfer them into the female. To examine how the additional steps of sperm transfer contribute to decreases in sperm quality, we assessed sperm viability (the proportion of live sperm) at each stage of mating and after different storage times in male and female reproductive organs in two damselfly species, Mnais pruinosa and Calopteryx cornelia. Viability of stored sperm in females was lower than that of male stores even just after copulation. Male sperm vesicles were not equipped to maintain sperm quality for longer periods than the internal reproductive organs. However, the sperm vesicles were only used for short-term storage; therefore, this process appeared unlikely to reduce sperm viability when transferred to the female. Males remove rival sperm prior to transfer of their own ejaculate using a peculiar-shaped aedeagus, but sperm removal by males is not always complete. Thus, dilution occurs between newly received sperm and aged sperm already stored in the female, causing lower viability of sperm inside the female than that of sperm transferred by males. If females do not remate, sperm viability gradually decreases with the duration of storage. Frequent mating of females may therefore contribute to the maintenance of high sperm quality.     

Female mating frequency in a wild population of scorpionflies (Panorpa germanica, Panorpidae, Mecoptera)

In this study, we investigated mating frequencies of female scorpionflies Panorpa germanica in the field using two different experimental approaches. First, the mating status of wild‐caught females was estimated on the basis of sperm numbers present inside their sperm storage organs. Secondly, the number of mating partners wild‐caught females must have had was inferred from mother‐offspring analyses of a polymorphic microsatellite locus. Our results suggest a very low mating frequency of wild female P. germanica. Consequently, the risk of sperm competition is rather low in this species. The relevance of female mate choice in relation to the low mating frequency is discussed.     

Sperm numbers,their storage and usage in the fly Dryomyza anilis

In the fly Dryomyza anilis females have two kinds of sperm storage organs: one bursa copulatrix and three spermathecae (two spermathecae with a common duct form the doublet, and the third is a singlet spermathecal unit). At the beginning of a mating the male deposits his sperm in the bursa copulatrix. After sperm transfer the male taps the female''s abdomen with his claspers. This behaviour has been shown to increase the male''s fertilization success. After mating, the female discharges large quantities of sperm before oviposition. To find out where the sperm remaining in the female are stored, I counted the number of sperm in the droplet and in the female''s sperm storage organs after different types of mating. I carried out three mating experiments. In experiment 1, virgin females were mated with one male and the matings were interrupted either immediately after sperm transfer or after several tapping sequences. The results show that during male tapping more sperm moved into the singlet spermatheca. In addition, the total number of sperm correlated with sperm numbers in all sperm storage organs, and male size was positively related to the number of sperm remaining in the bursa. In experiment 2, females mated with several males. The number of sperm increased with increasing number of matings only in the doublet spermatheca. No increase in the number of sperm in the singlet spermatheca during consecutive matings suggests that sperm were replaced or did not reach this sperm storage organ. In experiment 3, virgin females were mated with a single male and half of them were allowed to lay eggs. The experiment showed that during egglaying, females primarily used sperm from their singlet spermatheca. The results from the three experiments suggest that sperm stored in the singlet spermatheca is central for male fertilization success and male tapping is related to sperm storage in the singlet spermatheca. The different female''s sperm storage organs in D. anilis may have separate functions during sperm storage as well as during sperm usage.     

Sex peptide receptor is required for the release of stored sperm by mated Drosophila melanogaster females

The storage of sperm in mated females is important for efficient reproduction. After sperm are transferred to females during mating, they need to reach and enter into the site(s) of storage, be maintained viably within storage, and ultimately be released from storage to fertilize eggs. Perturbation of these events can have drastic consequences on fertility. In Drosophila melanogaster, females store sperm for up to 2 weeks after a single mating. For sperm to be released normally from storage, Drosophila females need to receive the seminal fluid protein (SFP) sex peptide (SP) during mating. SP, which binds to sperm in storage, signals through the sex peptide receptor (SPR) to elicit two other effects on mated females: the persistence of egg laying and a reduction in sexual receptivity. However, it is not known whether SPR is also needed to mediate SP’s effect on sperm release. By phenotypic analysis of flies deleted for SPR, and of flies knocked down for SPR, ubiquitously or in specific tissues, we show that SPR is required to mediate SP’s effects on sperm release from storage. We show that SPR expression in ppk⁺ neurons is needed for proper sperm release; these neurons include those that mediate SP’s effect on receptivity and egg laying. However, we find that SPR is also needed in the spermathecal secretory cells of the female reproductive tract for efficient sperm release. Thus, SPR expression is necessary in both the nervous system and in female reproductive tract cells to mediate the release of stored sperm.