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.

Ecological niche modeling and geographical distribution of pollinator and plants: A case study of Peponapis fervens (Smith, 1879) (Eucerini: Apidae) and Cucurbita species (Cucurbitaceae)

The bees of the Peponapes genus (Eucerini, Apidae) have a Neotropical distribution with the center of species diversity located in Mexico and are specialized in Cucurbita plants. which have many species of economic importance. such as squashes and pumpkins Peponapis fervens is the only species of the genus known from southern South America The Cucurbita species occurring in the same area as P fervens Include four domesticated species (C ficifolia, C maxima maxima, C moschata and C pepo) and one non-domesticated species (Cucurbita maxima andreana) It was suggested that C. in andreana was the original pollen source to P fervens, and this bee expanded its geographical range due to the domestication of Cucurbita The potential geographical areas of these species were determined and compared using ecological niche modeling that was performed with the computational system openModeller and GARP with best subsets algorithm The climatic variables obtained through modeling were compared using Cluster Analysis Results show that the potential areas of domesticated species practically spread all over South America The potential area of P fervens Includes the areas of C m andreana but reaches a larger area, where the domesticated species of Cucurbita also Occur The Cluster Analysis shows a high climatic similarity between P fervens and C. m. andreana Nevertheless. P fervens presents the ability to occupy areas with wider ranges of climatic variables and to exploit resources provided by domesticated species (C) 2009 Elsevier B V All rights reserved

Reproductive biology of six Brazilian Myrtaceae: is there a syndrome associated with buzz-pollination?

The floral phenology and reproductive biology of six sympatric arboreal Myrtaceae species were studied in the coastal plain forest (Ubatuba, Brazil, 44 degrees 48`W 23 degrees 22`S), from September 1999 to April 2002. Flowering started in the transition from the driest to the most humid season (Sep/Oct) and lasted until March. The sequence with which the species flowered each year was consistently the same. However, the timing of flowering onset, peak, end, and overlap differed from one year to another. Myrtaceae species were classified as xenogamic according to the pollen:ovule ratios, but two of them seem to present some degree of self-compatibility. Flowers of all species opened at sunrise and lasted for I day. Bombus morio (Apidae: Bombini) was the most common visitor followed by Melipona rufiventris (Apidae: Meliponini). Buzz pollination in Myrtaceae was common at the study area and seems to be related to bees` behaviour and to some aspects of flowers` morphology.

Ecological niche modeling and principal component analysis of Krameria Loefl. (Krameriaceae)

Krameria plants are found in arid regions of the Americas and present a floral system that attracts oil-collecting bees. Niche modeling and multivariate tools were applied to examine ecological and geographical aspects of the 18 species of this genus, using occurrence data obtained from herbaria and literature. Niche modeling showed the potential areas of occurrence for each species and the analysis of climatic variables suggested that North American species occur mostly in deserted or xeric ecoregions with monthly precipitation below 140 mm and large temperature ranges. South American species are mainly found in deserted ecoregions and subtropical savannas where monthly precipitation often exceeds 150 mm and temperature ranges are smaller. Principal Component Analysis (PCA) performed with values of temperature and precipitation showed that the distribution limits of Krameria species are primarily associated with maximum and minimum temperatures. Modeling of Krameria species proved to be a useful tool for analyzing the influence of the ecological niche variables in the geographical distribution of species, providing new information to guide future investigations. (C) 2011 Elsevier Ltd. All rights reserved.

Hybridization and introgression across different ploidy levels in the Neotropical orchids Epidendrum fulgens and E-puniceoluteum (Orchidaceae)

The hypothesis of gene flow between species with large differences in chromosome numbers has rarely been tested in the wild, mainly because species of different ploidy are commonly assumed to be reproductively isolated from each other because of instantaneous and strong postzygotic barriers. In this study, a broad-scale survey of molecular variation was carried out between two orchid species with different ploidy levels: Epidendrum fulgens (2n = 2x = 24 chromosomes) and Epidendrum puniceoluteum (2n = 4x = 52 chromosomes). To test the strength of their reproductive barriers, we investigated the distribution of genetic variation in sympatric and allopatric populations of these two species and conducted crossing experiments. Nuclear and plastid microsatellite loci were used to genotype 463 individuals from eight populations across the geographical range of both species along the Brazilian coastal plain. All six sympatric populations analysed presented hybrid zones, indicating that hybridization between E. fulgens and E. puniceoluteum is a common phenomenon. Bayesian assignment analysis detected the presence of F(1) and F(2) individuals and also signs of introgression, demonstrating a high potential for interspecific gene flow. Introgression occurs preferentially from E. fulgens to E. puniceoluteum. Pure parental individuals of both species display strong genotype-habitat associations, indicating that environment-dependent selection could be acting in all hybrid zones. This study suggests that hybridization and introgression are evolutionary processes playing a role in the diversification of Epidendrum and indicates the importance of investigations of hybrid zones in understanding reproductive barriers and speciation processes in Neotropical orchid species.

Genetic structure of Partamona helleri (Apidae, Meliponini) from Neotropical Atlantic rainforest

Stingless bees of the genus Partamona are distributed from southern Mexico to southern Brazil. This genus has been subject to different approaches to solve questions concerning general biology, taxonomy, systematics and biogeography, but population studies applying molecular techniques are inexistent. We analyzed the genetic structure of P. helleri across its geographic distribution along the coastal Atlantic tropical rainforest in Brazil. Ten mtDNA haplotypes were observed in 47 colonies of P. helleri of which some were exclusive and others shared among geographic sub-groups. Statistical analysis showed high genetic differentiation between geographic areas sampled. Fragmentation of the Atlantic forest during Pleistocene glaciations is discussed as a possible cause of the present haplotype distribution and frequency.

Isolation and characterization of 15 microsatellite loci in the stingless bee Plebeia remota (Apidae: Meliponini)

The destruction of Brazilian natural habitats has reduced bee populations and negative impacts of native flora pollination have been noticed. This work describes the isolation and characterization of microsatellite loci and evaluates them as molecular markers to study genetic variability of the stingless bee Plebeia remota. A microsatellite enriched genomic library was constructed and 15 primer pairs were designed for this species. The survey was conducted by analyzing 21 unrelated individuals. Genetic diversity indexes were calculated. The mean allelic richness was 6.3, the observed heterozygosity was 0.568, and the percentage of polymorphic loci was 93.33%. Also the primers were tested in cross-species amplification and showed promising results for P. droryana, P. emerina, P. lucii, P. meridionalis, P. pugnax, and P. saiqui. The microsatellite loci described here will be useful to evaluate genetic variability of stingless bees, and certainly will improve our knowledge about population dynamics especially in threatened environments.

Morphometrical, biochemical and molecular tools for assessing biodiversity. An example in Plebeia remota (Holmberg, 1903) (Apidae, Meliponini)

We see today many efforts to quantify biodiversity in different biomes. It is very important then to develop and to apply other methodologies that allow us to assess biodiversity. Here we present an example of application of three tools with this goal. We analyzed two populations of Plebeia remota from two distinct biomes that already showed several differences in morphology and behavior. Based on these differences, it has been suggested that the populations of Cunha and Prudentopolis do not represent a single species. In order to verify the existence or absence of gene flow between these two groups, we characterized the patterns of mtDNA through RFLP, the patterns of wing venation through geometric morphometry, and the cuticular hydrocarbons through gas chromatography-mass spectrometry. We used bees collected in these two locations and also from colonies which have being kept for around 9 years at Sao Paulo University. We found six different haplotypes in these specimens, of which three of them occurred exclusively in the population of Cunha and three only in the Prudentopolis population. The fact that the populations do not share haplotypes suggests no maternal gene flow between them. The two populations were differentiated by the pattern of the wing veins. They also had different mixtures of cuticle hydrocarbons. Furthermore it was shown that the colonies kept at the university did not hybridize. These two groups may constitute different species. We also show here the importance of using other methodologies than traditional taxonomy to assess and understand biodiversity, especially in bees.

Molecular Characterization of Propolis-Induced Cell Death in Saccharomyces cerevisiae

Propolis, a natural product of plant resins, is used by the bees to seal holes in their honeycombs and protect the hive entrance. However, propolis has also been used in folk medicine for centuries. Here, we apply the power of Saccharomyces cerevisiae as a model organism for studies of genetics, cell biology, and genomics to determine how propolis affects fungi at the cellular level. Propolis is able to induce an apoptosis cell death response. However, increased exposure to propolis provides a corresponding increase in the necrosis response. We showed that cytochrome c but not endonuclease G (Nuc1p) is involved in propolis-mediated cell death in S. cerevisiae. We also observed that the metacaspase YCA1 gene is important for propolis-mediated cell death. To elucidate the gene functions that may be required for propolis sensitivity in eukaryotes, the full collection of about 4,800 haploid S. cerevisiae deletion strains was screened for propolis sensitivity. We were able to identify 138 deletion strains that have different degrees of propolis sensitivity compared to the corresponding wild-type strains. Systems biology revealed enrichment for genes involved in the mitochondrial electron transport chain, vacuolar acidification, negative regulation of transcription from RNA polymerase II promoter, regulation of macroautophagy associated with protein targeting to vacuoles, and cellular response to starvation. Validation studies indicated that propolis sensitivity is dependent on the mitochondrial function and that vacuolar acidification and autophagy are important for yeast cell death caused by propolis.

A importância dos mecanismos comportamentais de resistência para a dinâmica populacional de abelhas Apis mellifera e o parasita Varroa destructor

Os ácaros ectoparasitas Varroa destructor, que parasitam as abelhas tornaram-se um problema global. Embora seja pouco provável que estes ácaros, por si só, provoquem a mortalidade das colmeias, eles desempenham um importante papel como vetor de muitas doenças virais. E estas doenças são identificados como algumas das mais importantes razões para a Desordem do Colapso das Colônias. Os efeitos da infestação do V.destructor são distintas em diferentes partes do mundo. Maiores mortalidades de colônias têm sido relatadas em colônias de abelhas européias (AE) em países da Europa, Ásia e América do Norte. No entanto, este ácaro está presente no Brasil já por muitos anos e não existem relatos de perdas em colônias das abelhas africanizadas (AA). Estudos realizados no México mostraram que alguns comportamentos de resistência ao ácaro Varroa - especialmente o grooming e o comportamento higiênico - são diferentes em cada uma das subespécie. Poderiam então esses mecanismos explicar por que as abelhas africanizadas são menos suscetíveis à Desordem do Colapso das Colônias? A fim de responder a esta pergunta, propomos um modelo matemático baseado em equações diferenciais, com o objetivo de analisar o papel desses mecanismos de resistência na saúde geral da colônia e na capacidade da colônia para enfrentar desafios ambientais.

BIOCHEMICAL VARIABILITY BETWEEN VENOMS FROM DIFFERENT HONEYBEE (APIS-MELLIFERA) RACES

1. The comparison of molecular exclusion cromatography profiles of venoms from sting apparatuses of Apis mellifera ligustica, Apis mellifera adansonii and Africanized honey-bees in Sephadex G-100 revealed both qualitative and quantitative differences.2. The venoms from A.m. ligustica and A.m. adansonii presented, respectively, three and two peaks characteristic of each sub-species, while Africanized honey-bee was characterized by the absence of eight peaks common to the former.3. The polypeptides with M(r) in the range from 100,000 to 7500 da correspond respectively to 62.0%, 66.6% and 68.7% of total proteins from the venon of A.m. ligustica, A.m. adansonii and Africanized honey-bees, while the peptidic fraction with M(r) range from 4100 to 2000 da corresponds to 11.4%, 32.4% and 10.2% of venom protein, respectively.

Entomofauna visitante de Stenolobium stans (Juss.) Seem (Bignoniaceae), durante seu período de floração

Realizaram-se estudos sobre as formas de reprodução de Stenolobium stans (Juss.) Seem e determinou-se a diversidade, freqüência e constância dos insetos visitantes nas flores em diferentes horas, durante quatro anos. As flores de S. stans começam a se abrir nas primeiras horas do dia entre 5 e 6h, com duração de 3 a 8h. Quando o estigma está receptivo, o pólen tem 90% de viabilidade. Além do pólen, a flor possui outros atrativos para os insetos visitantes, ou seja, os osmóforos responsáveis pelo odor adocicado, luz ultravioleta refletida e néctar com 25% de açúcar. A planta é autocompatível, reproduzindo-se por autogamia, geitonogamia ou xenogamia o que determina a necessidade de polinizadores externos e justifica ser a espécie vegetal em estudo uma séria invasora de campos e pastagens. Grande diversidade de insetos foi verificada visitando as flores, com predominância das abelhas. Os polinizadores foram Centris collaris Lepeletier, Bombus morio (Swederus), Eulaema nigrita Lepeletier e Epicharis sp. No meio rural houve menor incidência das espécies nativas do que no ambiente urbano, com predominância da abelha introduzida Apis mellifera L. Fatores ambientais, principalmente a temperatura, luminosidade, umidade relativa do ar e velocidade do vento, influenciaram a atividade forrageadora dos insetos.

Visitantes florais de Lagerstroemia speciosa Pers: (Lythraceae)

Studies were carried out with Lagerstroemia speciosa Pers. on floral reproductive systems, diversity and constancy of visiting insects at different hours of day, the behaviour of these insects at the flowers and the influence of these environmental factors in relation to their visits. The fenology, anthesis and others particularity of this vegetal species was studied. A great diversity of insects was verified visiting the flowers with the predominance of bees. The most frequent and constant species encountered were: Nannotrigona testaceicornis (Lepeletier, 1836) (40,2%), Tetragonisca angustula (Latreille, 1811) (16,9%), Apis mellifera Linnaeus, 1758 (11,8%), Plebeia droryana (Friese, 1900) (9,1 %) e Exomalopsis fulvofasciata (Smith, 1879) (8,5%). The blossoms possessis features of melittophily syndrome and diurnal anthesis. The environmental factors influence the insects foraging activity, mainly temperature, light, time of day, humidity and wind speed. The effective pollinators were the large insects like Bombus morio (Swederus, 1787), Bombus atratus (Franklin, 1913), Centris tarsata (Smith, 1874), Centris flavifrons Fabricius, 1775, Xylocopa suspecta Camargo & Moure, 1988, Xylocopa frontalis (Olivier, 1789) and Eulaema nigrita Lepeletier, 1841.

Visitantes florais de Erythrina speciosa Andr. (Leguminosae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)