Phylogenomics Controlling for Base Compositional Bias Reveals a Single Origin of Eusociality in Corbiculate Bees.

As increasingly large molecular data sets are collected for phylogenomics, the conflicting phylogenetic signal among gene trees poses challenges to resolve some difficult nodes of the Tree of Life. Among these nodes, the phylogenetic position of the honey bees (Apini) within the corbiculate bee group remains controversial, despite its considerable importance for understanding the emergence and maintenance of eusociality. Here, we show that this controversy stems in part from pervasive phylogenetic conflicts among GC-rich gene trees. GC-rich genes typically have a high nucleotidic heterogeneity among species, which can induce topological conflicts among gene trees. When retaining only the most GC-homogeneous genes or using a nonhomogeneous model of sequence evolution, our analyses reveal a monophyletic group of the three lineages with a eusocial lifestyle (honey bees, bumble bees, and stingless bees). These phylogenetic relationships strongly suggest a single origin of eusociality in the corbiculate bees, with no reversal to solitary living in this group. To accurately reconstruct other important evolutionary steps across the Tree of Life, we suggest removing GC-rich and GC-heterogeneous genes from large phylogenomic data sets. Interpreted as a consequence of genome-wide variations in recombination rates, this GC effect can affect all taxa featuring GC-biased gene conversion, which is common in eukaryotes.

Cladistic analysis of self-grooming indicates a single origin of eusociality in corbiculate bees (Hymenoptera: Apidae)

Behavioural traits have been used extensively in recent years as an important character source for making phylogenetic inferences. The phylogenetic positions of the members of the Apini subtribe are increasingly being debated, and new characters must be examined. We analysed the presence and absence of certain behavioural patterns, as well as the sequences of some of these patterns, to generate 79 characters. Eleven species comprised the ingroup, and Xylocopini comprised the outgroup. Parsimony analysis showed that the most parsimonious tree was (Euglossina(Bombina(Apina+Meliponina))). This topology is consistent with most studies that use morphological data and the few that use behavioural data, which suggests that advanced eusociality arose only once in a common ancestor of the clade Apina plus Meliponina; however, this hypothesis is inconsistent with our molecular data. Thus we considered behavioural, molecular, and morphological data and recovered the same topology, in which eusociality has a single origin in corbiculate bees.

A molecular phylogeny of the stingless bee genus Melipona (Hymenoptera: Apidae)

Stingless bees (Meliponini) constitute a diverse group of highly eusocial insects that occur throughout tropical regions around the world. The meliponine genus Melipona is restricted to the New World tropics and has over 50 described species. Melipona, like Apis, possesses the remarkable ability to use representational communication to indicate the location of foraging patches. Although Melipona has been the subject of numerous behavioral, ecological, and genetic studies, the evolutionary history of this genus remains largely unexplored. Here, we implement a multigene phylogenetic approach based on nuclear, mitochondrial, and ribosomal loci, coupled with molecular clock methods, to elucidate the phylogenetic relationships and antiquity of subgenera and species of Melipona. Our phylogenetic analysis resolves the relationship among subgenera and tends to agree with morphology-based classification hypotheses. Our molecular clock analysis indicates that the genus Melipona shared a most recent common ancestor at least similar to 14-17 million years (My) ago. These results provide the groundwork for future comparative analyses aimed at understanding the evolution of complex communication mechanisms in eusocial Apidae. (C) 2010 Elsevier Inc. All rights reserved.

Filogenia das abelhas corbiculadas (Apidae, Apinae): uso combinado de dados comportamentais de auto-limpeza e moleculares

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Africanized honey bees are efficient at detecting, uncapping and removing dead brood

The hygienic behavior of honey bees is based on a two-step process, including uncapping and removing diseased, dead, damaged, or parasitized brood inside the cell. We evaluated during periods of 1 h the time that hygienic and non-hygienic colonies of Africanized honey bees spend to detect, uncap and remove pin-killed brood using comb inserts with transparent walls placed in observation hives. We observed that hygienic colonies are significantly faster in detecting, uncapping and removing dead brood in the cells (P < 0.001).

Gender identification of five genera of stingless bees (Apidae, Meliponini) based on wing morphology

Currently, the identification of pollinators is a critical necessity of conservation programs. After it was found that features extracted from patterns of wing venation are sufficient to discriminate among insect species, various studies have focused on this structure. We examined wing venation patterns of males and workers of five stingless bee species in order to determine if there are differences between sexes and if these differences are greater within than between species. Geometric morphometric analyses were made of the forewings of males and workers of Nannotrigona testaceicornis, Melipona quadrifasciata, Frieseomelitta varia, and Scaptotrigona aff. depilis and Plebeia remota. The patterns of males and workers from the same species were more similar than the patterns of individuals of the same sex from different species, and the patterns of both males and workers, when analyzed alone, were sufficiently different to distinguish among these five species. This demonstrates that we can use this kind of analysis for the identification of stingless bee species and that the sex of the individual does not impede identification. Computer-assisted morphometric analysis of bee wing images can be a useful tool for biodiversity studies and conservation programs.

Differences between the quality of strawberries (Fragaria x ananassa) pollinated by the stingless bees Scaptotrigona aff. depilis and Nannotrigona testaceicornis

We investigated the success of two stingless bee species in pollinating strawberries in greenhouses. Three greenhouses and one open field area were used; one greenhouse had only strawberry plants (control), another (G1) had three colonies of Scaptotrigona aff. depilis and another (G2) had three colonies of Nannotrigona testaceicornis. In the open field area, the flowers could be visited by any bee. The total production of fruits was counted and a random sample (N = 100) from each area was used to measure weight, length, circumference, and achenes number (N = 5). The percentages of deformed strawberries were: 23% (no bees); 2% (greenhouses with bees) and 13% (open field). The strawberries from the greenhouse with N. testaceicornis and the open field were heavier than those from the greenhouses with no bees and with S. depilis. The fruit circumference was largest in the greenhouses with bees. The achenes number did not differ among the experimental areas. The strawberries produced in the greenhouses with stingless bees had more quality and greater commercial value than the fruits produced in the open field area and the greenhouse without bees. We conclude that stingless bees are efficient pollinators of strawberry flowers cultivated in greenhouses.

Trap-nesting bees (Hymenoptera: Apoidea) in forest fragments of the State of Sao Paulo, Brazil

We studied the community ecology of trap-nesting bees in two forest fragments of the State of Sao Paulo, Brazil, during two years, utilizing bamboo canes and tubes made of black cardboard as trap nests. The traps were inspected once a month with an otoscope. One hundred and fifteen nests were obtained at Estacao Ecologica de Paulo de Faria, Paulo de Faria (EEPF). These included nine species belonging to five genera and two families. At Santa Cecilia Farm (SCF), 12 species belonging to seven genera and three families built 392 nests. Natural enemies reared from nests of both areas included Hymenoptera, Diptera and Coleoptera. Species richness was similar between the areas but the communities differed considerably in species composition. The higher diversity found at EEPF was due to more even distribution of the species. No difference was observed between the numbers of nests built in each year in each area. Although the species richness was lower in the cool/dry season of both years at SCF, and in the first year at EEPF, the nesting frequencies did not differ between seasons for both the overall community but for each of the most abundant species. No annual fluctuation in the frequencies of nesting was observed. As temperature and precipitation were not found to be significantly different between the two years of study in each area, we concluded that climatic stability resulted in population stability.

Sequential hygienic behavior in Carniolan honey bees (Apis mellifera carnica)

We examined the sequence, order or steps of hygienic behavior (HB) from pin-killed pupae until the removal of them by the bees. We conducted our study with four colonies of Apis mellifera carnica in Germany and made four repetitions. The pin-killing method was used for evaluation of the HB of bees. The data were collected every 2 h after perforation, totaling 13 observations. Additionally, for one hygienic colony and another non-hygienic colony, individual analyses of each dead pupa were made at every observation, including all details, steps or sequences of HB. The bees recognize the cells containing dead pupae within 2 h after perforation, initially making a hole in the capping, which is the beginning of HB. Uncapping of the dead brood cell reached maximum values from 4 to 6 h after perforation; after 24 h, practically all cells were already uncapped. Another variable, called brood partially removed, was analyzed 4 h after perforation, after the cells had been perforated, which involved uncapping, followed by partial or total removal of the brood. Maximum values of brood partially removed were found 10 h after perforation, though such cells could be found up to 48 h after perforation. The most frequent sequence of events in both colonies was: capped cell -> punctured cell. brood partially removed -> empty cell. A new model of three pairs of recessive genes (uncapping u1, u2 and remover r) was proposed in order to explain the genetic control of the HB in Apis mellifera. We recommend evaluating HB 24 h after perforation and using a correction factor to compensate for control removal levels. We found a series of details of HB, which allow a study of how various factors may affect the sequence of the activities involved in HB and investigation of the genetics that controls this process.

Comparative study of the hygienic behavior of Carniolan and Africanized honey bees directed towards grouped versus isolated dead brood cells

In Apis mellifera, hygienic behavior involves recognition and removal of sick, damaged or dead brood from capped cells. We investigated whether bees react in the same way to grouped versus isolated damaged capped brood cells. Three colonies of wild-type Africanized honey bees and three colonies of Carniolan honey bees were used for this investigation. Capped worker brood cells aged 12 to 14 days old were perforated with the pin-killing method. After making holes in the brood cells, the combs were placed back into the hives; 24 h later the number of cleaned cells was recorded in areas with pin-killed and control brood cells. Four repetitions were made in each colony. Isolated cells were more frequently cleaned than grouped cells, though variance analysis showed no significant difference (P = 0.1421). Carniolan bees also were somewhat, though not significantly more hygienic than Africanized honey bees (P = 0.0840). We conclude that honey bees can detect and remove both isolated and grouped dead brood. The tendency towards greater hygienic efficiency directed towards grouped pin-killed brood may be a consequence of a greater concentration of volatiles emanating from the wounds in the dead pupae.

The first record on Brazilian stingless bees published 450 years ago by Hans Staden

Only a few decades after 1492, when Christopher Columbus arrived on a Caribbean island and Pedro Alvares Cabral claimed Brazil for Portugal in 1500, a German mercenary gave the first description of stingless bees in 1557. He got to know them when he was imprisoned for months by an anthropophagous tribe in the coastal region of Santos, today in the State of Sao Paulo. This rather short but nevertheless extremely exact record on stingless bees is hidden in the first book on Brazil. Three species and important aspects of their life history were treated. This early description has been completely overlooked by bee scientists until now. My note intends to close this evident gap.

Africanized honey bees more efficiently convert protein diets into hemolymph protein than do Carniolan bees (Apis mellifera carnica)

The superiority of Africanized over European honey bees in tropical and subtropical regions of the New World is both well documented and poorly understood. As part of an effort to try to understand the process by which the displacement of European bees occurred, we examined the ability of these two types of bees and of hybrids between the two to convert natural and artificial diets into usable protein. Newly emerged bees from colonies of tropically adapted Africanized and temperate-origin Carniolan bees and first-generation hybrids between the two were caged and fed artificial and natural protein diets for six days to determine whether there were differences in their ability to use these diets. The Africanized bees developed significantly higher protein levels in the hemolymph than did the Carniolan bees. The difference was 31% when the bees were fed bee bread (37.5 and 28.56 mu g protein/mu L hemolymph, respectively). The hybrids developed protein levels intermediate between the two parental types. These were approximately 10 times the levels found in bees fed with sucrose alone. Superior food conversion efficiency of the Africanized bees may be one of the reasons for their superiority both in the wild and for beekeeping in tropical and subtropical Latin America.

Male sleeping aggregations of solitary oil-collecting bees in Brazil (Centridini, Tapinotaspidini, and Tetrapediini; Hymenoptera: Apidae)

Males of solitary bees usually spend the night in clusters on small branches of plants, cavities and flowers. The individuals usually return to the same location each evening during their life, exhibiting site fidelity to a particular plant. We report on the sleeping roosts of the males of some oil-collecting bees of the genera Centris, Paratetrapedia, Lanthanomelissa, Monoeca, and Tetrapedia, as well as the host plants. We discuss the role of the male clusters to the associated plants.

Chemical analyses confirm a rare case of seed dispersal by bees

It has been suggested that dispersal of seeds of Coussapoa asperifolia magnifolia could have endozoochoric dispersal by frugivorous birds and monkeys because the fruits are red when ripe, or exozoochoric dispersal, because the exocarp is mucilaginous and sticky. However, our field observations showed only stingless bees collecting the exocarp with seeds of C. asperifolia magnifolia, which are used for building and repairing their nests, from which the plants sprout. This paper aimed to determine the fruit chemical composition, since we postulated that C. asperifolia magnifolia is neither consumed by birds nor monkeys due to being very sticky and apparently resinous. Apolar extract analyses revealed the fruits are not resinous but extremely rich in waxes ( mainly esterified triglycerides), and polar extract analyses revealed the sugar content to be close to the sensorial minimum level. This probably accounts for why only stingless bees are seen visiting fruits and dispersing seeds.

Pheromone paths attached to the substrate in meliponine bees: helpful but not obligatory for recruitment success

In contrast to marking of the location of resources or sexual partners using single-spot pheromone sources, pheromone paths attached to the substrate and assisting orientation are rarely found among flying organisms. However, they do exist in meliponine bees (Apidae, Apinae, Meliponini), commonly known as stingless bees, which represent a group of important pollinators in tropical forests. Worker bees of several Neotropical meliponine species, especially in the genus Scaptotrigona Moure 1942, deposit pheromone paths on substrates between highly profitable resources and their nest. In contrast to past results and claims, we find that these pheromone paths are not an indispensable condition for successful recruitment but rather a means to increase the success of recruiters in persuading their nestmates to forage food at a particular location. Our results are relevant to a speciation theory in scent path-laying meliponine bees, such as Scaptotrigona. In addition, the finding that pheromone path-laying bees are able to recruit to food locations even across barriers such as large bodies of water affects tropical pollination ecology and theories on the evolution of resource communication in insect societies with a flying worker caste.