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。     

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。     

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     

蝇蛹金小蜂的交配行为及雄蜂交配次数对雌蜂繁殖的影响

多数昆虫能够进行多次交配,随寄生蜂雄蜂交配次数的增加,雄蜂体内精子减少,雌蜂获得的精子数量减少,产下更多的单倍体卵,发育为雄性后代;一些单寄生性的寄生蜂雌蜂一生仅能够交配1次。描述了蝇蛹金小蜂雌雄蜂的交配行为,探讨了雄蜂交配次数对雌蜂后代产量等的影响以及雌蜂的可交配次数。结果表明,交配过程包括求偶、交尾前期、交尾和交尾后期;雄蜂已交配的次数并不能够显著影响其配偶的寿命、产卵期和后代总数量,但显著影响到其配偶的雌、雄后代数量和性比。随雄蜂交配次数的增加,与之交配的雌蜂的后代雄性百分比显著增大,雌蜂在产卵期内更早地出现较多雄性后代,体内精子不足的现象更加明显。无论已交配的蝇蛹金小蜂雌蜂在产卵期中是否出现精子不足,均不能再次完成交配。     

一种斑翅果蝇寄生蜂的交配行为观察及雄虫交配次数对繁殖的影响

【目的】明确一种斑翅果蝇寄生蜂Trichopria drosophilae的交配行为以及雌雄蜂的交配次数对后代的影响。【方法】在室内对斑翅果蝇寄生蜂的交配行为进行观察,记录该蜂在交配行为中所出现的求偶行为、交尾前期行为、交尾行为和交尾完毕的动作及持续时间。测定寄生蜂各交配次数下的雌蜂寿命、后代出蜂总量和后代性比。【结果】寄生蜂的交配过程包括以下几个阶段:求偶,雄蜂逐渐靠近雌虫、追逐雌蜂并震动翅膀,直至爬上雌虫背部整个过程持续(50.47±85.01)s。交尾前期,雄虫头部从雌蜂的两触角中间伸出,并将触角从雌虫触角两侧向中间有规律的触碰雌虫触角,直至雌蜂打开生殖孔,整个过程持续(43.73±13.97)s。交尾,雄蜂将雄性交配器插入雌性生殖孔整个过程持续(36.28±11.03)s。交尾后期,雌虫左右甩动腹部2~3次,主动与雄虫分离整个过程持续(8.95±3.40)s。观察结果显示,雌虫一生只能交配一次,雄蜂一生交配次数在(16.54±1.37)次,最多达到19次(N=10),雄虫的交配次数对与之交配的雌蜂的寿命和后代总量无显著影响。雄蜂的交配次数对雌性后代的性比有显著影响。【结论】该寄生蜂的交配过程主要分为雄虫求偶、交尾前期、交尾、交尾后期等阶段。求偶和交尾前期阶段主要行为是雄虫追逐雌蜂、爬上雌虫背部并用触角摩擦雌虫触角。交尾阶段主要行为是雄蜂交配器插入雌性生殖孔。交尾后期主要行为是雌蜂将雄蜂甩开,是雌虫唯一主动发起的行为。随着雄蜂交配次数增加,雌蜂后代雌/雄性比降低。     

蜂群中不同亚家庭的工蜂寿命分析

在蜂群中,蜂王与多个雄蜂自然交尾,形成不同亚家庭.为了研究不同亚家庭中工蜂寿命是否有差异,我们以西方蜜蜂Apis mellifera L.为实验材料,随机从一群自然群中取270只刚孵化的工蜂,单独饲养于有蜜粉脾小蜂箱内,每天将自然死亡个体取出标记.并利用3对微卫星进行个体基因型分析,通过Matesoft软件划分亚家庭,然后分析了各亚家庭工蜂的自然寿命及生存曲线.结果表明:实验蜂群由12个亚家庭组成,其中2个亚家庭工蜂寿命与其他亚家庭存在显著差异(P＜0.05).     

中华蜜蜂精巢和卵巢差异表达基因分析

[目的]在蜂群中,雄蜂与蜂王都有着发育完全的性腺,但两者达到性成熟的时间却是不同步的.本研究旨在探究中华蜜蜂Apis cerana cerana雄蜂与蜂王生殖腺的基因表达差异.[方法]利用Illumina测序技术对中华蜜蜂雄蜂精巢与蜂王卵巢转录组差异表达基因(differentially expressed genes...     

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。     

蜜蜂蜂王质量及其性状表现影响因素研究进展

作为重要的传粉昆虫,蜜蜂蜂群损失现象受到广泛关注。研究表明,蜂王问题是导致蜂群损失的主要因素之一。蜂王是蜂群中唯一雌性生殖器官发育完全的个体,是维持蜂群存续的关键。蜂王质量决定了蜂群的群势以及生产性能,对蜂群的发展和存活至关重要。本文详细介绍了表征蜂王质量的相关指标及其在蜂王选育过程中的应用,深入论述了幼虫日龄、营养、交尾、环境温度、病虫害以及农药等相关因素对蜂王质量及其性状表现的影响,以期为蜂王的培育和使用以及蜜蜂资源的保护和利用提供参考。     

蜜蜂群内生殖分工体系的形成及其维持

本文对蜜蜂群内生殖分工体系的形成及其维持机制进行综述。蜜蜂群体具有完善的劳动分工(包括生殖分工)体系,蜂王垄断生殖权力,而工蜂生殖器官发育不完全,在蜂王信息素和幼虫信息素的作用下产卵受到抑制。蜂王的多雄交配机制降低了群内个体间的亲缘关系,但也促进了工蜂间相互监督机制的形成。工蜂间的相互监督,结合蜂王和幼虫信息素对工蜂卵巢发育的影响,解决了蜂王与工蜂、工蜂与工蜂间的生殖利益冲突,保障了蜂群内的生殖分工体系,提高了群体效率,维护了蜂群的真社会性。     

Estimation of intracolonial worker relationship in a honey bee colony (Apis mellifera L.) using DNA fingerprinting

Summary The number and frequencies of subfamilies in a honey bee colony were determined by DNA fingerprinting. Queen and brood samples were taken from three colonies with artificially inseminated queens and from one colony with a naturally mated queen. UsingHae III restriction enzyme and (GATA)₄ oligonucleotide, the number of subfamilies in the colonies with artificially inseminated queens corresponded with the number of drones used for insemination. In the colony with the naturally mated queen, 12 subfamilies were found in a random sample of 104 workers. Considering that subfamily frequencies range from 1 to 26%, introcolonial worker relationship was estimated to be 0.328, corresponding to a genetical effective number of 6.4 matings.     

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。     

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.     

Paternity and maternity frequencies in Apis mellifera sicula

Summary: Honey bee queens have been shown to mate with a high number of males, but the evolutionary advantage of this high degree of polyandry is still unclear. Mating data from a number of different Apis species and subspecies are needed to help explain polyandry in honey bees. Pupae of four colonies of Apis mellifera sicula from Sicily were genotyped on three polymorphic microsatellite loci. The genotypes of the queens and fathering drones from these colonies were deduced from the genotypes of the pupae. We found no evidence for polygyny, at least we can exclude more than one functional queen, even super-sister queens, if maternity contributions are equal. The four queens mated with at least 5 to 12 (mean: 9.3 - 3.0 SE) drones. We estimate the error in our determination of the mating frequency that is caused by limited genetic resolution of the marker loci to be less than 1 mating given that Hardy-Weinberg assumptions are satisfied. However, the drones the single queens mated with may be a non-random sample of the whole population, so that detection error may be more severe. The average pedigree relatedness among workers within the colonies was estimated to be 0.341. These results are within the range of those found in other A. mellifera subspecies and Apis species except A. dorsata. We speculate that mating frequency may be positively correlated with drone density.     

Queen reproductive state modulates pheromone production and queen-worker interactions in honeybees

The mandibular glands of queen honeybees produce a pheromone that modulates many aspects of worker honeybee physiology and behavior and is critical for colony social organization. The exact chemical blend produced by the queen differs between virgin and mated, laying queens. Here, we investigate the role of mating and reproductive state on queen pheromone production and worker responses. Virgin queens, naturally mated queens, and queens instrumentally inseminated with either semen or saline were collected 2 days after mating or insemination. Naturally mated queens had the most activated ovaries and the most distinct chemical profile in their mandibular glands. Instrumentally inseminated queens were intermediate between virgins and naturally mated queens for both ovary activation and chemical profiles. There were no significant differences between semen- and saline-inseminated queens. Workers were preferentially attracted to the mandibular gland extracts from queens with significantly more activated ovaries. These studies suggest that the queen pheromone blend is modulated by the reproductive status of the queens, and workers can detect these subtle differences and are more responsive to queens with higher reproductive potential. Furthermore, it appears as if insemination substance does not strongly affect physiological characteristics of honeybee queens 2 days after insemination, suggesting that the insemination process or volume is responsible for stimulating these early postmating changes in honeybee queens.     

Mating frequency and mating system of the polygynous ant, Leptothorax acervorum

Multiple mating by queens (polyandry) and the occurrence of multiple queens in the same colony (polygyny) alter patterns of relatedness within societies of eusocial insects. This is predicted to influence kin-selected conflicts over reproduction. We investigated the mating system of a facultatively polygynous UK population of the ant Leptothorax acervorum using up to six microsatellite loci. We estimated mating frequency by genotyping 79 dealate (colony) queens and the contents of their sperm receptacles and by detailed genetic analysis of 11 monogynous (single-queen) and nine polygynous colonies. Results indicated that 95% of queens were singly mated and 5% of queens were doubly mated. The corrected population mean mating frequency was 1.06. Parentage analysis of adults and brood in 17 colonies (10 monogynous, 7 polygynous) showed that female offspring attributable to each of 31 queens were full sisters, confirming that queens typically mate once. Inbreeding coefficients, queen-mate relatedness of zero and the low incidence of diploid males provided evidence that L. acervorum sexuals mate entirely or almost entirely at random. Males mated to queens in the same polygynous colony were not related to one another. Our data also confirmed that polygynous colonies contain queens that are related on average and that their workers had a mixed maternity. We conclude that the mating system of L. acervorum involves queens that mate near nests with unrelated males and then seek readoption by those nests, and queens that mate in mating aggregations away from nests, also with unrelated males.     

羽化和性成熟时中华蜜蜂蜂王和雄蜂转录组分析

【目的】为了系统了解中华蜜蜂Apis cerana cerana蜂王和雄蜂转录组特征,丰富蜜蜂转录组数据信息。【方法】本研究利用高通量测序的方法分别检测中华蜜蜂蜂王和雄蜂刚出房、性成熟时期以及性成熟期蜂王生殖系统和雄蜂生殖系统之间转录组表达差异。【结果】经过测序获得质量值不低于20的碱基比例（Q20）均高于90%;所有reads组装成90 839个unigenes,平均长度1 549 bp;基于5个数据库(NR,Swiss Prot,GO,COG和KEGG)进行比对,共有45 112个unigenes被注释。差异基因表达分析发现,与刚出房时相比,性成熟的蜂王和雄蜂均在表皮蛋白、细胞色素P450、气味结合蛋白等家族基因上存在显著差异表达,而且这些差异表达基因与蜜蜂生长发育和性成熟过程中蜜蜂骨骼发育、生殖系统发育、嗅觉发育等方面有关;性成熟蜂王与性成熟雄蜂之间以及它们生殖系统之间在气味结合蛋白基因方面存在显著差异。【结论】结果表明,中华蜜蜂在性成熟过程中,体内大量基因的表达发生了变化。这些结果揭示了中华蜜蜂性成熟发育的整体基因表达特征,在得到大量转录组unigene序列的同时,获得了一批与蜜蜂性成熟有关的基因序列,为深入开展中华蜜蜂生长发育与繁殖研究提供了丰富的数据资源。     

Phenotypic variation in susceptibility of honey bees,Apis mellifera,to infestation by tracheal mites,Acarapis woodi

A laboratory bioassay was used to study phenotypic differences in susceptibility of honey bees,Apis mellifera L., to tracheal mites,Acarapis woodi Rennie. Significantly different infestation frequencies were found in bees from 23 colonies containing queens that were instrumentally inseminated with single drones. Queens and drones originated from a closed population composed of commercial stock from various areas of the United States.Mites were randomly distributed with respect to right and left prothoracic tracheae. Tracheae containing mites were no more or less attractive to migrating mites than non-infested tracheae. The same quantity of progeny per female was produced in tracheae containing 1–3 mites. Female mites apparently do not migrate a second time after egg laying begins.The degree of phenotypic variation suggests that selection of honey bees for tracheal mite resistance is feasible.     

Queen promiscuity lowers disease within honeybee colonies

Most species of social insects have singly mated queens, but in some species each queen mates with numerous males to create a colony with a genetically diverse worker force. The adaptive significance of polyandry by social insect queens remains an evolutionary puzzle. Using the honeybee (Apis mellifera), we tested the hypothesis that polyandry improves a colony's resistance to disease. We established colonies headed by queens that had been artificially inseminated by either one or 10 drones. Later, we inoculated these colonies with spores of Paenibacillus larvae, the bacterium that causes a highly virulent disease of honeybee larvae (American foulbrood). We found that, on average, colonies headed by multiple-drone inseminated queens had markedly lower disease intensity and higher colony strength at the end of the summer relative to colonies headed by single-drone inseminated queens. These findings support the hypothesis that polyandry by social insect queens is an adaptation to counter disease within their colonies.     

Resistance to Varroa destructor (Mesostigmata: Varroidae) when mite-resistant queen honey bees (Hymenoptera: Apidae) were free-mated with unselected drones.

This study demonstrated (1) that honey bees, Apis mellifera L, can express a high level of resistance to Varroa destructor Anderson & Trueman when bees were selected for only one resistant trait (suppression of mite reproduction); and (2) that a significant level of mite-resistance was retained when these queens were free-mated with unselected drones. The test compared the growth of mite populations in colonies of bees that each received one of the following queens: (1) resistant--queens selected for suppression of mite reproduction and artificially inseminated in Baton Rouge with drones from similarly selected stocks; (2) resistant x control--resistant queens, as above, produced and free-mated to unselected drones by one of four commercial queen producers; and (3) control--commercial queens chosen by the same four queen producers and free-mated as above. All colonies started the test with approximately 0.9 kg of bees that were naturally infested with approximately 650 mites. Colonies with resistant x control queens ended the 115-d test period with significantly fewer mites than did colonies with control queens. This suggests that beekeepers can derive immediate benefit from mite-resistant queens that have been free-mated to unselected drones. Moreover, the production and distribution of these free-mated queens from many commercial sources may be an effective way to insert beneficial genes into our commercial population of honey bees without losing the genetic diversity and the useful beekeeping characteristics of this population.