Methods for species delimitation in bumblebees (Hymenoptera,Apidae, Bombus): towards an integrative approach

Delimitation of closely related species is often hindered by the lack of discrete diagnostic morphological characters. This is exemplified in bumblebees (genus Bombus). There have been many attempts to clarify bumblebee taxonomy by using alternative features to discrete morphological characters such as wing shape, DNA, or eco‐chemical traits. Nevertheless each approach has its own limitations. Recent studies have used a multisource approach to gather different lines of speciation evidence in order to draw a strongly supported taxonomic hypothesis in bumblebees. Yet, the resulting taxonomic status is not independent of selected evidence and of consensus methodology (i.e. unanimous procedure, majority, different weighting of evidence). In this article, we compare taxonomic conclusions for a group of taxonomically doubtful species (the Bombus lapidarius‐group) obtained from the four commonly used lines of evidence for species delimitation in bumblebees (geometric morphometric of wing shape, genetic differentiation assessment, sequence‐based species delimitation methods and differentiation of cephalic labial gland secretions). We ultimately aim to assess the usefulness of these lines of evidence as components of an integrative decision framework to delimit bumblebee species. Our results show that analyses based on wing shape do not delineate any obvious cluster. In contrast, nuclear/mitochondrial, sequence‐based species delimitation methods, and analyses based on cephalic labial gland secretions are congruent with each other. This allows setting up an integrative decision framework to establish strongly supported species and subspecies status within bumblebees.     

Male behaviour in two bumblebees, Bombus nevudensis auricomus and B. griseicollis (Hymenoptera: Apidae)

The mating system of the bumblebees Bombus nevadensis and B. griseicollis involves apparent territorial defence of perch sites near or within visually conspicuous trees. Non–contact chases between perch owners and intruders are rare in B. nevadensis and more common in B. griseicollis. The perches of B. nevadensis are in open areas whereas those of B. griseicoilis are within trees or in sites shaded by trees. The same locations may be occupied on 2 different years. Individual males may return to favoured perches over periods ranging from 2 to 24 days. Males of both species scent–mark spots near their perches early in the morning and then scan visually for passing females for as much as 7–8 h daily. The landmark–based mating system of these bumblebees is similar to that of some other insects and closely resembles the pure dominance (lek) mating system of certain vertebrates. Some hypotheses for the evolution of diversity in male mating tactics among bumblebee species are presented.     

Proboscis length and resource utilization in two Uruguayan bumblebees: Bombus atratus Franklin and Bombus bellicosus Smith (Hymenoptera: Apidae)

Bumblebees (Bombus sp.) are eusocial insects with an annual life cycle whose use as pollinator of crops has gained great importance in modern agriculture. Several authors have found that resource use in Bombus species is usually based on the correlation between the proboscis length of the bumblebees and the corolla depth of the flowers. The aim of this study was to determine proboscis length of Bombus atratus and B. bellicosus, two Uruguayan bumblebees, and verify the resource exploitation testing two cultivated species, the red clover and the bird's foot trefoil. Bumblebee foraging activity was recorded in two culture conditions: in a red clover and bird's foot trefoil mixed meadow, and in contiguous plots of these legumes, and the proboscis length of collected foragers was determined. Both species of bumblebees exploited red clover and bird's foot trefoil although they did it in different proportions in all instances tested. The results indicated that the choice of the resources in B. atratus and B. bellicosus was influenced by their proboscis length. Bombus atratus has a longer proboscis and preferably visited red clover, possibly obtaining nectar easier and faster than B. bellicosus, which has a shorter proboscis. Bombus bellicosus used both resources without any clear preference.     

Density-dependent and frequency-dependent selection by bumblebees Bombus terrestris (L.) (Hymenoptera: Apidae)

The behaviour of bumblebee workers foraging on arrays of artificial flowers of two colour morphs was observed. Experiments were conducted on arrays of varying morph frequencies and at three different total flower densities. Bumblebees consistently showed a preference for the commonest colour morph, and this behaviour was not significantly affected by changing density. In contrast, frequency-independent preferences changed significantly with density. At low densities, there was a strong bias towards the more conspicuous colour, whereas at higher densities there was no overall colour bias. Flight distances between flowers decreased significantly at high density. Bumblebees also visited flowers of similar colours sequentially, but this behaviour was not density-dependent. It is suggested that as densities increase, there is an increased probability that bumblebees detect yellow flowers, which were probably less conspicuous compared with blue flowers, and that this might be caused by changes in flight speed with flight distance. Where there is a positive relationship between pollinator visitation and the relative fitness of a floral morph, the observed behaviour would induce positive frequency-dependent selection on a plant population with two corolla colour morphs on which the bumblebees were foraging, which would result in stabilizing selection for a single corolla colour, irrespective of density. There was no indication that rare colour morphs would be preferred at high density. The probability of different corolla colour morphs going to fixation would, however, be affected by density.     

A method for year-round rearing of cuckoo bumblebees (Hymenoptera: Apoidea: Bombus subgenus Psithyrus)

Summary. The study of the social interactions between host bumblebees and their inquiline species requires rearing them. Here we provide a simple method for rearing of cuckoo bumblebees (Bombus subgenus Psithyrus) in the laboratory. Two bumblebee cuckoo-host systems are used: Bombus (Psithyrus) vestalis hosted by B. (Bombus) terrestris and B. (Psithyrus) sylvestris hosted by B. (Pyrobombus) pratorum. First, the invasion of the Psithyrus female is performed in species-specific host colonies containing approximately 10 young workers. On average, a parasitized colony of B. terrestris produced 90 ± 9 young males and 21 ± 3 young females of B. vestalis, whereas a parasitized colony of B. pratorum produced 16 ± 2 young males and 5 ± 1 young females of B. sylvestris. One week after emergence, Psithyrus virgin females are placed in a flight cage exposed to natural light, with Psithyrus males of other colonies. After successful mating, the females are isolated and overwintered in a cold room at 4°C for 2–4 months. This method enables a year-round and mass breeding of Psithyrus species to facilitate studies of these rare species.     

Functional morphology of the visual system and mating strategies in bumblebees (Hymenoptera,Apidae, Bombus)

Bumblebee mating behaviour intrigued Darwin over 150 years ago and has elicited much interest since. Despite our increasing knowledge about male mating strategies, we still know little about the signals and cues involved, the sender‐receiver relationship and the functional adaptations of the involved sensory systems. Although several authors have noticed a close relationship between the mating system and male‐specific elaboration of the visual system, studies on the functional adaptations have yet to be conducted. We investigated the functional morphology of the compound eyes in 11 species of Bombus from various subgenera in detail. Of these, four species (Bombus confusus, Bombus melaleucus, Bombus mendax, Bombus niveatus) have enlarged male eyes, characterized by a higher number of ommatidia compared with worker eyes and a frontal zone with enlarged facets, which is likely to be associated with improved spatial resolution and contrast sensitivity. In these species, perching mating strategies are found. In contrast, males of species that patrol scent routes (Bombus hortorum, Bombus lapidarius, Bombus pascuorum, Bombus pratorum, Bombus soroeensis, Bombus terrestris, Bombus wurflenii) show no distinct eye adaptations, and their eye morphology closely resembles that of the workers. A phylogenetic analysis of male eye structure indicates that enlarged males eyes may have been the ancestral state in Bombus. © 2014 The Linnean Society of London     

Do social parasitic bumblebees use chemical weapons? (Hymenoptera,Apidae)

The bumblebee Bombus (Psithyrus) norvegicus Sp.-Schn. is an obligate social parasite of B. (Pyrobombus) hypnorum L. Behavioural observations indicated that nest-invading B. norvegicus females may use allomones to defend themselves against attacking host workers. However, so far no defensive chemicals used by social parasitic bumblebee females have been identified. We analysed volatile constituents of the cuticular lipid profile of B. norvegicus females. Furthermore, we performed electrophysiological studies and behavioural experiments in order to identify possible chemical weapons. Coupled gas chromatography-electroantennography showed 15 compounds to trigger responses in antennae of the host workers. Using gas chromatography–mass spectrometry, the main compound among the cuticular volatiles of B. norvegicus females was found to be dodecyl acetate. A corresponding mixture of synthetic volatiles as well as pure dodecyl acetate showed a strong repellent effect on starved host workers. B. norvegicus females use dodecyl acetate to repel attacking B. hypnorum workers during nest usurpation and subsequently during colony development. Dodecyl acetate is the first repellent allomone identified in bumblebees.     

Exocrine gland secretions of virgin queens of five bumblebee species (Hymenoptera: Apidae, Bombini)

Secretions of three different glands (mandibular gland, labial gland, and Dufour's gland) of virgin queens of five bumblebee species (Bombus lucorum, B. lapidarius, B. hypnorum, B. pascuorum, and B. terrestris) were analysed. Around 200 compounds were identified in the secretions. The compositions of the secretions of labial and mandibular glands were species-specific. Dufour's gland of all species produced mainly hydrocarbons, both saturated and unsaturated, the proportions of which differed quantitatively between the species studied.     

The first record of the rare oligolectic bumblebee Bombus gerstaeckeri Morawitz (Hymenoptera: Apidae: Bombini) lrom Ukraine

The oligolectic bumblebee Bombus gerstaeckeri Morawitz has been recorded in the Ukrainian Carpathians for the first time during field research from 2002 to 2007. Queens and workers of the species have been observed foraging on two Aconitum spp. within 540 m and 1400 m altitude. The additional forage resource of this species, that is Salvia glutinosa L., has been recorded at low height. Three competitor bumblebees, not confined only to Aconitum spp. have been observed foraging on monkshoods: B. hortorum (L.), B. pascuorum (Scopoli) and B. wurflenii Radoszkowski.     

The bumblebees of the subgenus Subterraneobombus: integrating evidence from morphology and DNA barcodes (Hymenoptera,Apidae, Bombus)

Although bumblebees have received a lot of attention, some taxonomic problems have persisted for many years. One particularly obdurate case has been the species of the subgenus Subterraneobombus. We revise the bees of this subgenus by integrating evidence from both morphology and, for a 5% subsample, from DNA (cytochrome c oxidase subunit 1, CO1) barcodes from pinned museum specimens. We apply a reciprocal illumination procedure: (1) taxa recognized previously from morphology are used to stratify samples for DNA subsampling; (2) DNA barcodes from these subsamples are used to recognize groups of phylogenetically related specimens; and (3) for these groups, we re‐examine morphological characters in order to recognize and diagnose species. A total of 3854 specimens from 1535 samples from across the geographic range of the subgenus throughout the Holarctic and northern Oriental regions are identified to 11 species. This includes one species newly recognized from Mongolia, Bombus mongolensis Williams sp. nov. Taxon concepts are modified substantially for four species, seven lectotypes are designated, and four new synonyms are recognized. The prevailing usage of Bombus distinguendus is maintained as valid by designating Bombus elegans as a nomen oblitum and designating B. distinguendus as a nomen protectum. Identification keys and colour‐pattern diagrams are provided, and geographic distributions, elevational ranges, and phenological activity periods are described to characterize the species. An estimate of the biogeographic history is reconstructed with dispersal–vicariance analysis. In this study, DNA barcode data have been a cost‐effective source of additional characters for diagnosing groups of specimens. The barcode data contributed directly to recognizing the one new species, of which females remain difficult to identify from morphology alone. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 163 , 813–862.     

Fauna of bumblebees (Hymenoptera: Apidae: Bombus Latreille) in the mainland part of Arkhangelsk Region,NW Russia

The focus of this study is to present bumblebee fauna of Arkhangelsk Region (north-western Russia). This research is based on the museum collections and materials collected by the authors. Collecting localities and data of faunistic records are given in the species list. We include 34 species in the fauna of bumblebees of Arkhangelsk Region. The regional fauna is dominated by Transpalaearctic species. Bumblebee fauna in the study region has low specificity. The recent distribution of bumblebee species in the study region is a result of post-glacial immigration.     

Genetic structure of Korean populations of bumblebees Bombus ignitus (Hymenoptera: Apidae) as revealed by microsatellite markers

Bumblebee Bombus ignitus, which is indigenous to Korea, Japan, and China, has been recognized as a valuable pollinator for both crops and wild plants. Bombus ignitus has now become commercially important as a pollinator because of its use in the agricultural industry, particularly for greenhouse pollination. For long‐term management and effective conservation of B. ignitus, an understanding of the genetic structure of its naturally occurring populations is practically important. In this study, the genetic structure among the five populations of B. ignitus in South Korea was assessed using nine microsatellite loci. These markers showed high levels of genetic variability, with the number of alleles ranging from 6 to 22 (mean = 13.4) and the frequency of the most common allele ranging from 0.11 to 0.66. Only the Sabuk (SB) population showed a genetic signature of a recent bottleneck, which was further supported by the lowest level of allelic richness (AR) (mean AR = 3.944). Genetic differentiation was highly significant among all pairs of populations (P < 0.001) across the nine microsatellite markers, suggesting a lack of gene flow among those populations. Interestingly, F_ST (and R_ST and D_est) values were always greater for the Taebaek population than for the four remaining populations. The phylogenetic analysis showed evidence supporting our hypothesis that the Taebaek population is genetically more divergent than the other populations. Overall, our results suggest that the Korean populations of B. ignitus might have undergone geographic isolation and have become highly separated spatially from one another, thereby having a limited range of migration among their local habitats.     

Wolbachia在熊蜂中的双重感染

Wolbachia是一类广泛存在于节肢动物体内细胞质遗传的细菌,它可以通过诱导产雌孤雌生殖、引起细胞质不亲和、遗传雄性的雌性化、雄性致死和增强生殖力等作用方式引起其寄主生殖行为的改变.本文以16S rDNA为标记检测了3种熊蜂不同组织(头,胸,足,卵巢或雄外生殖器)的Wolbachia感染.其中明亮熊蜂Bombus l...     

Bombus impatiens (Hymenoptera: Apidae): an alternative to Apis mellifera (Hymenoptera: Apidae) for lowbush blueberry pollination.

The pollination effectiveness of the commercially reared bumble bee Bombus impatiens Cresson, was compared in field studies to the honey bee, Apis mellifera L., for lowbush blueberry, Vaccinium angustifolium Ait. A preliminary study indicated that B. impatiens had potential as an alternative pollinator. In a 3-yr study, percentage fruit set, percentage harvested berries, berry weight, and seeds per berry were compared in blueberry fields stocked at 7.5 A. mellifera hives per hectare to 5, 7.5, or 10 B. impatiens colonies per hectare. Percentage of harvested berries (yield) was significantly higher in fields stocked with B. impatiens at 10 colonies per hectare. No other parameters measuring pollinator effectiveness were significantly different at 5, 7.5, or 10 colonies per hectare. Flower handling time was significantly faster for B. impatiens and it more frequently collected blueberry pollen. All parameters of pollinator effectiveness were similar for B. impatiens, A. mellifera, and native wild bees in a follow-up study. Overall, B. impatiens was a suitable alternative to A. mellifera.     

Frequency-dependent selection by pollinators: mechanisms and consequences with regard to behaviour of bumblebees Bombus terrestris (L.) (Hymenoptera: Apidae)

Bombus terrestris , a typical pollinating insect species, was offered artificial flowers of two different corolla colours with the same sucrose solution reward in an array. Common colours were significantly preferred, and the strength of the frequency-dependent response increased as a result of learning. There were also frequency-independent biases towards blue flowers, probably because blue flowers appeared more conspicuous to bumblebees than yellow flowers, and the degree of preference for blue was greater when flowers had low nectar rewards. Flower-to-flower movements by individual bumblebees between flowers were non-random, were biased to movements within the same flower colour, and were also dependent on morph frequency. The mechanisms governing flower selection in bumblebees are discussed. Pollinators foraging similarly in a natural situation would induce positive frequency-dependent selection, assortative mating, and directional selection on different corolla colour morphs of the plant population being visited, resulting in stabilizing selection for a single flower colour.     

Atypical secretions of the male cephalic labial glands in bumblebees: the case of Bombus (Rhodobombus) mesomelas Gerstaecker (Hymenoptera, Apidae)

In bumblebees, the male secretion of the cephalic labial gland is species-specific. It is highly involved in the nuptial behavior, acting as a sexual attracting pheromone. Therefore, it is also used to accurately identify the species. In contrast to this common scheme, the secretions of Bombus mesomelas are found to be strongly reduced and do not include the most volatile compounds that are present in the secretions of all the other studied bumblebee species. These secretions correspond to cuticular hydrocarbons that can be found in all bumblebee species. This was also the case for another bumblebee species from the same Rhodobombus subgenus: Bombus pomorum. This atypical composition of the male cephalic labial gland secretions seems to indicate that, at least for these two species of Rhodobombus, these secretions are not used to attract virgin females from a long distance, as it is the case for all the other bumblebee species studied.     

小峰熊蜂访花偏爱性

传粉昆虫在访花时,通常会表现出对某一类型花的偏爱性。本研究利用人工制作的大小、颜色、形态和气味不同的9种类型的花来研究小峰熊蜂Bombus hypocrita的访花偏爱性。结果表明：当增加花的大小、形态、气味等附加特征数时,小峰熊蜂的偏爱性程度与花朵附加特征数有显著相关性（P<0.01）。当花朵颜色由2种增加到4种,熊蜂对紫色花的偏爱性程度降低,但花朵颜色的种类与小峰熊蜂的访花偏爱性没有相关性（P>0.05）,花颜色的种类对熊蜂访紫色花的偏爱性影响不大。大小为5 cm的紫花被访次数（108±9次）明显高于大小为3 cm的紫花被访次数（40±4次）（P<0.01）,说明熊蜂明显喜欢访大花瓣的紫花。完全盛开的紫花被访次数（129±13次）显著高于刚绽放的花被访次数（26±3次）（P<0.01）,说明熊蜂喜欢访盛开的紫花。柠檬味的紫花被访次数（63±8次）明显低于草莓味的紫色花被访次数（88±2次）（P<0.05）,说明熊蜂喜欢访草莓味的花朵。     

Comparison and examination of Bombus occidentalis and Bombus impatiens (Hymenoptera: Apidae) in tomato greenhouses

Experiments were conducted in commercial tomato, Lycopersicon esculentum Miller (Solanaceae), greenhouses to compare the relative foraging effort of two bumble bee species, Bombus occidentalis Greene and Bombus impatiens Cresson, to examine interspecific competition between B. occidentalis and B. impatiens, and to determine whether bumble bee colonies grew to their full population potential in commercial tomato greenhouses. B. impatiens colonies had more brood and workers and made more foraging trips per hour than B. occidentalis colonies. However, B. impatiens returned to the colony without pollen loads and left their colonies without dropping off their pollen loads more frequently than B. occidentalis greenhouse colonies. Our data also suggest that the presence of B. impatiens had a detrimental effect on B. occidentalis populations. Furthermore, B. occidentalis colonies did not grow to their full population potential in tomato greenhouses, with fewer workers in greenhouse colonies than in colonies placed outside in a natural environment, or in colonies that were physically enclosed and protected from external mortality. Together, this study suggests that B. impatiens is a better pollinator than B. occidentalis. It also shows that unknown factors are limiting the size of B. occidentalis colonies in tomato greenhouses.     

Reduced cephalic labial glands in the male bumblebees of the subgenus Rhodobombus Dalla Torre (Hymenoptera: Apidae: Bombus Latreille)

Until now, all males of bumblebees are known to attract conspecific females by marking places with a chemical signal secreted by their cephalic labial gland. The specific combination of patrolling flight and scentmarking is of outmost importance to bumblebees as it is their main species specific recognition system. We report here the lack of that recognition system in species of the subgenus Rhodobombus by comparing the morphology and histology of the cephalic labial glands of Bombus (Rhodobombus) mesomelas with those of a well known species, B. (Bombus) terrestris (L.) The cephalic labial glands are much smaller in B. mesomelas than in other bumblebees species and most likely non-functional. This morphology is also observed in B. pomorum and B. brodmanni which also belong to the subgenus Rhodobombus. Our morphological observations are consistent with the chemical analyses of the secretions in B. mesomelas and B. pomorum which are very limited and of most unusual composition for a bumblebee. In addition, whereas other species are thought to use their barbae mandibularis to spread their secretion onto the substrate, these structures are absent in all Rhodobombus. All these observations would mean that the males of Rhodobombus do not attract females from far away by the use of their cephalic labial gland secretions.