Genetic diversity and population structure of two Euglossini bee species in a host-parasite relationship.

In the current study, two euglossine species, Exaerete smaragdina and Eulaema nigrita, a cleptoparasite bee and its host, respectively, were used as models to: (i) access the genetic diversity and population structure of both species, sampled along a wide latitudinal range of Atlantic Forest, where the distribution of El. nigrita and Ex. smaragdina co-occurs; (ii) investigate the evolutionary history of these species through the Atlantic Forest, and in a wider scenario, to examine the evolutionary history of these species across others forest domains. Analyses involved males of El. nigrita and Ex. smaragdina sampled through Brazilian territory, including 19 sites in the Atlantic Forest. Bayesian Skyline Plot (BSP) was used to infer possible climate oscillations on population of both species over time. The BSP revealed stability in effective population size for both species in most of the Plio-Pleistocene period. However, BSP results aligned to the starlike configuration in the haplotype network, neutrality test, and population diversity patterns indicated population expansion of the two species during the late Pleistocene. Our findings suggest areas of potential refugia to the climatic oscillations of the Pleistocene in the Atlantic Forest in the Brazilian states of Espírito Santo for El. nigrita and Pernambuco for Ex. smaragdina.

Isolation and characterization of microsatellite markers in two species of the neotropical Epicharis (Hymenoptera, Apidae, Centridini) genus and cross-amplification in related species.

Genetics can provide invaluable tools for management and conservation of bee populations, which are declining worldwide. Among these tools, microsatellite are very useful molecular markers for population analyses. The aim of this study was to isolate and characterize microsatellites for Epicharis (Anepicharis) dejeanii and Epicharis (Epicharis) nigrita, two Neotropical species of solitary bees, both exhibiting the habit of nesting in aggregations. Microsatellite loci were identified from two enriched genomic libraries. The characterization and analysis of loci were carried out using 35 females of E. dejeanii and 34 of E. nigrita. In total, we report the development of 12 microsatellite loci for E. dejeanii and 13 for E. nigrita. For E. dejeanii, all loci were polymorphic, the number of alleles per locus ranged from 2 to 12, averaging 8.7 and, observed and expected heterozygosity were 0.485 (range 0.229-0.857) and 0.633 (range 0.288-0.843), respectively. For E. nigrita, only nine out of 13 loci amplified were polymorphic, the number of alleles per locus ranged from 2 to 12, averaging 5.5. For this species, the observed and expected heterozygosity were 0.440 (range 0.118-0.676) and 0.545 (range 0.167-0.814), respectively. Cross-amplification of primers was successful in other Centridini species. The two sets of loci described for E. dejeanii and E. nigrita species are polymorphic and informative and show promising applicability for both population genetic approaches and relatedness on these and other Centridini species.

Electric eels galore: microsatellite markers for population studies

Fourteen novel microsatellite loci are described and characterized in two species of electric eels, Electrophorus variiand E. voltaifrom floodplains and rivers of the Amazon rainforest. These loci are polymorphic, highly informative, and have the capacity to detect reliable levels of genetic diversity. Likewise, the high combined probability of paternity exclusion value and low combined probability of genetic identity value obtained demonstrate that the new set of loci displays suitability for paternity studies on electric eels. In addition, the cross-amplification of electric eel species implies that it may also be useful in the study of the closely related E. electricus, and to other Neotropical electric fishes (Gymnotiformes) species as tested herein.(AU)

Catorze novos loci microsatélites são descritos e caracterizados em duas espécies de poraquês, Electrophorus varii e E. voltai de planícies alagadas e rios da floresta amazônica. Esses loci são polimórficos, altamente informativos e têm a capacidade de detectar níveis confiáveis de diversidade genética. Da mesma forma, o alto valor de exclusão de paternidade combinado com a baixa probabilidade de identidade genética demonstra que o novo conjunto de loci exibe adequação para estudos de paternidade em poraquês. Além disso, a amplificação cruzada de espécies de peixes elétricos implica que também pode ser útil no estudo da espécie intimamente relacionada E. electricus, e de outras espécies de peixes elétricos neotropicais (Gymnotiformes).(AU)

Genetic diversity and population structure of Brycon nattereri (Characiformes: Bryconidae): a Neotropical fish under threat of extinction

Brycon nattereri is an endangered Neotropical fish reported along conserved stretches of the upper Paraná, Tocantins and São Francisco rivers. Populations of this species have been very rare in some Paraná River sub basins. This study analyzes the genetic diversity and population structure of B. nattereri in a restricted area of occurrence recently identified in upper Paraná River basin. Seven microsatellite loci and 497 bp of D-Loop mitochondrial region were examined in 92 individuals from four points along the area of occurrence. Both molecular markers indicated a single population distributed along a stretch of the river approximately 80 km long. Although some of the data suggest an ancient bottleneck, current levels of genetic diversity (H E = 0.574 and h = 0.616) were similar to those of other species of the genus Brycon. The results suggest that the population of B. nattereri has been able to maintain satisfactory levels of genetic diversity, in spite of the small area of occurrence. These data have highlighted an important conservation area and action may prove essential to improve the quality of the environment, and especially the water and riparian plant life, if the area is to be managed and conserved efficiently.(AU)

Brycon nattereri é um peixe Neotropical ameaçado de extinção reportado para trechos conservados dos rios Paraná, Tocantins e São Francisco. Populações desta espécie têm sido muito raras em algumas sub-bacias do rio Paraná. Este estudo analisou a diversidade genética e a estrutura populacional de B. nattereri em uma área de ocorrência restrita recentemente identificada na bacia do alto rio Paraná. Sete locos microssatélites e 497 pb da região mitocondrial D-Loop foram examinados para 92 indivíduos de quatro pontos ao longo da área de ocorrência. Ambos os marcadores moleculares indicaram uma única população distribuída em um trecho de aproximadamente 80 km do rio. Embora alguns dados tenham sugerido um antigo gargalo genético, os atuais níveis de diversidade genética (H E = 0,574, h = 0,616) foram similares aos de outras espécies do gênero Brycon. Estes resultados sugerem que a população de B. nattereri tem mantido níveis satisfatórios de diversidade genética, apesar da pequena área de ocorrência. Estes dados destacaram uma importante área de conservação e ações podem melhorar a qualidade do ambiente, especialmente para a vida aquática e mata ciliar, se a área for eficientemente manejada e conservada.(AU)

Reappraisal of the systematics of Microglanis cottoides (Siluriformes, Pseudopimelodidae), a catfish from southern Brazil.

The southern region of Brazil is characterized by high species diversity and endemism of freshwater fishes distributed across geographically isolated river basins. Microglanis cottoides has a widespread range across these river basins and occurs in sympatry with other endemic species of the genus (e.g. M. cibelae, M. eurystoma, and M. malabarbai). Herein we tested the monophyly of M. cottoides and presented for the first time information about the molecular phylogeny of species in the genus. The results suggest that M. cottoides currently forms a non-monophyletic group which includes populations endemic to the Uruguay River basin that are more closely related to M. malabarbai, and excludes M. cibelae, found to be nested within M. cottoides. Based on an integrative approach using morphological and molecular data, we propose M. cibelae as a junior synonym of M. cottoides, and the populations of the Uruguay River basin previously assigned to M. cottoides in fact belong to M. malabarbai. Our molecular phylogeny shows that M. cottoides is sister to M. parahybae, which is also a coastal species, and M. malabarbai is sister of M. garavelloi, both endemic to inland river basins. The time-calibrated phylogeny indicates that the separation between inland and the coastal clades occurred in the Tertiary period, and that the species within the coastal basins diverged in the Pliocene, which overlaps with the diversification times estimated for the two inland species as well. This pattern of diversification corroborates some previous studies with other fishes from the same region.

Genetic Diversity of the Endangered Neotropical Cichlid Fish (Gymnogeophagus setequedas) in Brazil.

Gymnogeophagus setequedas is a rare and rheophilic species of tribe Geophagini, considered endangered in Brazilian red lists. Its previously known geographical distribution range was the Paraná River basin, in Paraguay, and a tributary of the Itaipu Reservoir in Brazil. Since its description no specimens have been collected in the original known distribution area. However, recent records of G. setequedas in the lower Iguaçu River, in a region considered highly endemic for the ichthyofauna, extended the known geographical distribution and may represent one of the last remnants of the species. The aim of this study was to estimate the genetic diversity and population structure of G. setequedas, using microsatellite markers and mitochondrial haplotypes, in order to test the hypothesis of low genetic diversity in this restricted population. Muscular tissue samples of 86 specimens were obtained from nine locations in the Lower Iguaçu River basin, between upstream of the Iguaçu Falls and downstream of the Salto Caxias Reservoir. Seven microsatellites loci were examined and a total of 120 different alleles were obtained. The number of alleles per locus (NA) was 17.429, effective alleles (NE) 6.644, expected heterozygosity (HE) 0.675, observed (HO) heterozygosity 0.592, and inbreeding coefficient (FIS) 0.128. Twelve haplotypes in the D-Loop region were revealed, with values of h (0.7642) and π (0.00729), suggesting a large and stable population with a long evolutionary history. Thus, both molecular markers revealed high levels of genetic diversity and indicated the occurrence of a single G. setequedas population distributed along a stretch of approximately 200 km. The pattern of mismatch distribution was multimodal, which is usually ascribed to populations in demographic equilibrium. Nevertheless, the construction of a new hydroelectric power plant, already underway between the Salto Caxias Reservoir and Iguaçu Falls, could fragment this population, causing loss of genetic diversity and population decline, and for this reason it is necessary to maintain the Iguaçu River tributaries and downstream area from the Lower Iguaçu Reservoir free of additional dams, to guarantee the survival of this species.

Comparative analysis of adaptive and neutral markers of Drosophila mediopunctata populations dispersed among forest fragments.

Comparison of adaptive and neutral genetic markers is a valuable approach to characterize the evolutionary consequences of populations living in environments threatened by anthropogenic disturbances, such as forest fragmentation. Shifts in allele frequencies, low genetic variability, and a small effective population size can be considered clear signs of forest fragmentation effects (due to genetic drift) over natural populations, while adaptive responses correlate with environmental variables. Brazilian Atlantic Forest had its landscape drastically reduced and fragmented. Now, several forest remnants are isolated from each other by urban and crop areas. We sampled Drosophila mediopunctata populations from eight forest remnants dispersed on two adjacent geomorphological regions, which are physiognomic and climatically quite distinct. Microsatellite data of inversion-free chromosomes (neutral genetic marker) indicate low structuration among populations suggesting that they were panmictic and greatly influenced by gene flow. Moreover, significant differences in chromosomal inversion frequencies (adaptive genetic marker) among populations and their correlations with climatic and geographical variables indicate that genetic divergence among populations could be an adaptive response to their environment. Nonetheless, we observed a significant difference in inversion frequencies of a population in two consecutive years that may be associated with edge and demographic effects. Also, it may be reflecting seasonal changes of inversion frequencies influenced by great temperature variation due to edge effects. Moreover, the forest fragment size does not affect genetic variation of neutral markers. Our data indicate that despite oscillations in chromosomal inversion frequencies, D. mediopunctata populations from Brazilian Atlantic Forest and their divergence may be driven by adaptive factors to local differences, perhaps because it is a small flying insect easily carried by the wind increasing its migration rates.

Development and characterization of fifteen polymorphic microsatellite loci in Bryconamericus aff. iheringii (Teleostei: Characidae) and cross-amplification in related Characidae species

Data on 15 novel microsatellite loci from the Neotropical fish Bryconamericus aff. iheringii are presented here. Analyses of 32 individuals from four different streams revealed 192 different alleles, ranging from four to 32 alleles per locus (mean of 12.8 per locus). Observed and expected heterozygosities ranged from 0.094 to 0.813 and 0.205 to 0.952, respectively. These loci showed high polymorphic information content and will be a resource for genetic studies of B. aff. iheringii. Furthermore, several loci also amplified other small Neotropical Characidae (Piabarchus stramineus and Piabina argentea) and should be useful for these species.(AU)

Um total de quinze novos locos microssatélites é aqui apresentado para o pequeno peixe Neotropical Bryconamericus aff. iheringii. A análise de 32 indivíduos provenientes de quatro ribeirões diferentes revelou 192 alelos diferentes, variando de quatro a 32 alelos por loco (média de 12,8 por loco), e heterozigozidades observada e esperada variando de 0,094 a 0,813 e 0,205 a 0,952, respectivamente. O conjunto de locos obtido mostrou alto conteúdo de informação polimórfica e bom potencial para estudos genéticos de B. aff. iheringii, além disso vários locos amplificaram para outras espécies de pequenos Characidae neotropicais (Piabarchus stramineus and Piabina argentea).(AU)

Changes in Orchid Bee Communities Across Forest-Agroecosystem Boundaries in Brazilian Atlantic Forest Landscapes.

Deforestation has dramatically reduced the extent of Atlantic Forest cover in Brazil. Orchid bees are key pollinators in neotropical forest, and many species are sensitive to anthropogenic interference. In this sense understanding the matrix permeability for these bees is important for maintaining genetic diversity and pollination services. Our main objective was to assess whether the composition, abundance, and diversity of orchid bees in matrices differed from those in Atlantic forest. To do this we sampled orchid bees at 4-mo intervals from 2007 to 2009 in remnants of Atlantic Forest, and in the surrounding pasture and eucalyptus matrices. The abundance, richness, and diversity of orchid bees diminished significantly from the forest fragment toward the matrix points in the eucalyptus and pasture. Some common or intermediate species in the forest areas, such as Eulaema cingulata (F.) and Euglossa fimbriata Moure, respectively, become rare species in the matrices. Our results show that the orchid bee community is affected by the matrices surrounding the forest fragments. They also suggest that connections between forest fragments need to be improved using friendly matrices that can provide more favorable conditions for bees and increase their dispersal between fragments.

Mitochondrial DNA diversity of orchid bee Euglossa fimbriata (Hymenoptera: Apidae) populations assessed by PCR-RFLP.

Euglossa fimbriata is a euglossine species widely distributed in Brazil and occurring primarily in Atlantic Forest remnants. In this study, the genetic mitochondrial structure of E. fimbriata from six Atlantic Forest fragments was studied by RFLP analysis of three PCR-amplified mtDNA gene segments (16S, COI-COII, and cyt b). Ten composite haplotypes were identified, six of which were exclusive and represented singleton mitotypes. Low haplotype diversity (0.085-0.289) and nucleotide diversity (0.000-0.002) were detected within samples. AMOVA partitioned 91.13% of the overall genetic variation within samples and 8.87% (phi(st) = 0.089; P < 0.05) among samples. Pairwise comparisons indicated high levels of differentiation among some pairs of samples (phi(st) = 0.161-0.218; P < 0.05). These high levels indicate that these populations of E. fimbriata, despite their highly fragmented landscape, apparently have not suffered loss of genetic variation, suggesting that this particular population is not currently endangered.

Eufriesea violacea (Blanchard) (Hymenoptera: Apidae): an orchid bee apparently sensitive to size reduction in forest patches.

Eufriesea violacea (Blanchard) is a very seasonal euglossine species, more frequently found in the southern and southeastern regions of Brazil. A number of studies have revealed large variations in the abundance of males of this species present in Atlantic Forest remnants throughout both regions. In this paper, we report variations in the abundance of E. violacea males sampled in several forest patches of different sizes (ranging from 10 to 580 ha), and we propose that this species is possibly sensitive to the reduction in size of forest remnants. Surveys were carried out in nine forest remnants of Atlantic rainforest located in northern Paraná State, southern Brazil. Male euglossine bees were collected with an entomological net when visiting scent-baits, between 10:00 am and 1:00 pm, from October to December of 2001 and 2006. A total of 360 E. violacea males were captured in the nine forest fragments studied. The number of bees attracted to scent baits in each forest patch varied from zero to 261. A very high association (r = 0.993) was detected between the forest patch size and the visitation rate of E. violacea males at different sites, with the highest mean number of males visiting baits/sampling (43.5) being observed for bees from the largest forest remnant. Although alternative hypothesis should not be discharged for the decline in the abundance or absence of E. violacea in small forest patches, our results indicate that populations of this euglossine species need larger forest areas for existing.

Eufriesea violacea (Blanchard) (Hymenoptera: Apidae): an orchid Bee apparently sensitive to size reduction in forest patches / Eufriesea violacea (Blanchard) (Hymenoptera: Apidae): uma abelha das orquídeas aparentemente sensível à redução no tamanho dos fragmentos florestais

Eufriesea violacea (Blanchard) is a very seasonal euglossine species, more frequently found in the southern and southeastern regions of Brazil. A number of studies have revealed large variations in the abundance of males of this species present in Atlantic Forest remnants throughout both regions. In this paper, we report variations in the abundance of E. violacea males sampled in several forest patches of different sizes (ranging from 10 to 580 ha), and we propose that this species is possibly sensitive to the reduction in size of forest remnants. Surveys were carried out in nine forest remnants of Atlantic rainforest located in northern Paraná State, southern Brazil. Male euglossine bees were collected with an entomological net when visiting scent-baits, between 10:00 am and 1:00 pm, from October to December of 2001 and 2006. A total of 360 E. violacea males were captured in the nine forest fragments studied. The number of bees attracted to scent baits in each forest patch varied from zero to 261. A very high association (r = 0.993) was detected between the forest patch size and the visitation rate of E. violacea males at different sites, with the highest mean number of males visiting baits/sampling (43.5) being observed for bees from the largest forest remnant. Although alternative hypothesis should not be discharged for the decline in the abundance or absence of E. violacea in small forest patches, our results indicate that populations of this euglossine species need larger forest areas for existing.

Eufriesea violacea (Blanchard) é uma espécie de Euglossini bastante sazonal, de ocorrência mais frequente no Sul e Sudeste do Brasil. Vários estudos revelaram grandes variações na abundância de machos de E. violacea em remanescentes de Mata Atlântica dessas duas regiões brasileiras. Neste artigo, relatamos variações na abundância de machos de E. violacea de vários fragmentos florestais de tamanhos distintos (variando de 10 a 580 ha), e sugerimos que a espécie seja provavelmente sensível à redução no tamanho da área do fragmento florestal. As amostragens foram realizadas em nove remanescentes florestais de Mata Atlântica, localizados no Norte do Paraná, Sul do Brasil. Machos euglossíneos foram coletados com rede entomológica durante suas visitas às iscas-odores, entre 10:00h e 13:00h, de outubro a dezembro de 2001 e 2006. O total de 360 machos de E. violacea foi coletado nos fragmentos estudados. O número de abelhas atraídas às iscas em cada área variou de zero a 261 indivíduos. O tamanho do fragmento foi proporcionalmente relacionado (r = 0,993) à taxa de visitação de machos de E. violacea nas diferentes áreas, sendo que o maior número médio de machos (43,5) por amostragem foi observado para abelhas do fragmento florestal maior. Embora hipóteses alternativas para o declínio na abundância ou ausência de E. violacea em fragmentos florestais menores não devam ser descartadas, esses resultados indicam que populações de E. violacea necessitam de áreas florestais maiores para sua sobrevivência.

Genetic diversity of Hypostomus ancistroides (Teleostei, Loricariidae) from an urban stream

In this study, random amplified polymorphic DNA (RAPD) markers were applied to analyze the genetic diversity of samples of the Neotropical catfish Hypostomus ancistroides, collected from four sites (S1, S2, S3 and S4) along an urban stream in Southern Brazil. The 11 primers used in RAPD analysis amplified 147 loci, 76 (51.7 percent) of which were polymorphic. The proportions of polymorphic loci observed in the four samples were: 29.93 percent (S1), 31.97 percent (S2), 23.81 percent (S3) and 38.77 percent (S4). The average heterozygosity within sampling localities ranged from 0.1230 to 0.1526 and unbiased genetic distances ranged from 0.0253 to 0.0445. AMOVA partitioned 90.85 percent of the total variation within samples and 9.15 percent among samples. Excepting for the sample pair S1-S2 (phiST = 0.02784; p > 0.05), all others pairwise phiST values were significantly greater than zero, indicating moderate genetic differentiation among catfish samples from four localities. The relative low levels of genetic variation detected in all groups studied could be related to different factors, including the sedentary habit of these fish, which can be eroding the genetic variation of H. ancistroides from each locality.

[Bee diversity in Tecoma stans (L.) Kunth (Bignoniaceae): importance for pollination and fruit production]. / Diversidade de abelhas em Tecoma stans (L.) Kunth (Bignoniaceae): importância na polinização e produção de frutos.

Tecoma stans (L.) Kunth is an exotic plant in Brazil, commonly distributed in urban areas, which is considered an invasive species in crop and pasture areas. In this study, the floral biology and the behavior of bees in flowers of T. stans from three urban areas in southeastern Brazil were investigated. In all study sites, T. stans was an important food resource to the Apoidea to 48 species of bees. Centris tarsata Smith and Exomalopsis fulvofasciata Smith (Hymenoptera: Apidae) were the effective pollinators more abundant, while Scaptotrigona depilis Moure (Hymenoptera: Apidae) was the more frequent robber species. The most part of T. stans visitors (87.5%) exploited exclusively nectar, which varied in sugar concentration depending on the day period and flower phase. In all flower stages, higher averages of nectar concentration (26.4% to 32.7%) occurred from 10 am to 2 pm. The presence of osmophore in the petals and protandry were detected. In two urban areas the number of visitors varied significantly during the day. The greatest abundance of pollinators occurred when pollen availability was higher and flowers showed receptive stigma, which could be contributing to the reproductive success of T. stans. The results indicate that the production of fruits increased in plants that received a higher number of effective pollinators.

Diversidade de abelhas em Tecoma stans (L.) Kunth (Bignoniaceae): importância na polinização e produção de frutos / Bee diversity in Tecoma stans (L.) Kunth (Bignoniaceae): importance for pollination and fruit production

Tecoma stans (L.) Kunth é uma espécie vegetal introduzida no Brasil, comumente encontrada em áreas urbanas e considerada invasora de ambientes cultivados e áreas de pastagem. Neste trabalho foram estudados aspectos da biologia foral, diversidade e comportamento de coleta das abelhas em flores de T. stans em três áreas urbanas no Sudeste do Brasil. Nas três áreas, T. stans mostrou-se importante fonte de recursos alimentares utilizados 48 espécies de abelhas. Centris tarsata Smith e Exomalopsis fulvofasciata Smith (Hymenoptera: Apidae) foram os polinizadores efetivos mais abundantes e Scaptotrigona depilis Moure (Hymenoptera: Apidae) a espécie pilhadora mais freqüente. A grande maioria (87,5 por cento) dos visitantes de T. stans coletou exclusivamente néctar, recurso que variou em concentração durante o dia e nas diferentes fases da flor. Em todos os estágios florais, as maiores concentrações médias de néctar ocorreram entre 10:00h e 14:00h. Detectou-se a presença de osmóforos nas pétalas e protandria. Em duas áreas estudadas o número de visitantes variou significativamente durante o dia. A maior abundância de polinizadores ocorreu nos períodos de maior disponibilidade de pólen e estigmas receptivos, fato esse que pode ser determinante para o sucesso reprodutivo de T. stans. Os resultados indicam ainda maior produção de frutos em plantas que receberam o número maior de polinizadores efetivos.

Tecoma stans (L.) Kunth is an exotic plant in Brazil, commonly distributed in urban areas, which is considered an invasive species in crop and pasture areas. In this study, the floral biology and the behavior of bees in flowers of T. stans from three urban areas in southeastern Brazil were investigated. In all study sites, T. stans was an important food resource to the Apoidea to 48 species of bees. Centris tarsata Smith and Exomalopsis fulvofasciata Smith (Hymenoptera: Apidae) were the effective pollinators more abundant, while Scaptotrigona depilis Moure (Hymenoptera: Apidae) was the more frequent robber species. The most part of T. stans visitors (87.5 percent) exploited exclusively nectar, which varied in sugar concentration depending on the day period and flower phase. In all flower stages, higher averages of nectar concentration (26.4 percent to 32.7 percent) occurred from 10 am to 2 pm. The presence of osmophore in the petals and protandry were detected. In two urban areas the number of visitors varied significantly during the day. The greatest abundance of pollinators occurred when pollen availability was higher and flowers showed receptive stigma, which could be contributing to the reproductive success of T. stans. The results indicate that the production of fruits increased in plants that received a higher number of effective pollinators.

Evaluation of the RAPD profiles from different body parts of Euglossa pleosticta Dressler male bees (Hymenoptera: Apidae, Euglossina)

In the current literature, information is scarce on which part of the adult insect body is suitable for isolation of genomic DNA for genetic analysis based on DNA-markers. In this study, we evaluated RAPD profiles generated from total genomic DNA isolated from distinct body parts (head, legs, thorax + wings and abdomen) of 12 males of Euglossa pleosticta Dressler. From the total of bands analyzed, 9.0 percent did not show reproducibility. Percent variations of bands in each body segment were: 1.1 percent (head); 0.4 percent (legs); 0.8 percent (thorax/wings) and 6.7 percent (abdomen). The much higher variation (chi2one sample = 10.27; df = 1; P < 0.01) in the RAPD profiles obtained by using DNA isolated from abdomen of the euglossine males suggests that this body part of adult insects should be avoided in DNA extraction procedures. Conversely, the low variation among the RAPD profiles obtained from amplifications of genomic DNA extracted from head, legs and thorax/wings indicates that all these body parts of male bees are equally useful and secure for using in isolation and amplification procedures of total genomic DNA.

Na literatura atual, são escassas as informações sobre qual a parte do corpo do inseto adulto é mais adequada para a extração de DNA genômico para estudos de análises genéticas baseadas em marcadores de DNA. Neste estudo, foram analisados os perfis de RAPD produzidos a partir da amplificação do DNA genômico extraído de partes distintas do corpo (cabeça, pernas, tórax + asas e abdome) de 12 machos de Euglossa pleosticta Dressler. Do total de bandas analisadas, 9,0 por cento não mostraram repetibilidade. As porcentagens de variação de bandas em relação às diferentes partes do corpo das abelhas foram: 1,1 por cento (cabeça); 0,4 por cento (pernas); 0,8 por cento (tórax/asas) e 6,7 por cento (abdome). A maior variação observada (chi2para uma amostra = 10,27; gl = 1; P < 0,01), nos perfis eletroforéticos de RAPD, produzidos nas amplificações do DNA extraído do abdome de machos euglossíneos sugere que essa parte do corpo de insetos adultos deve ser evitada em procedimentos de extração de DNA. De modo diverso, a baixa variação entre os perfis de RAPD obtidos a partir das amplificações de DNA genômico extraído da cabeça, pernas e tórax/asas das abelhas indica que todas essas partes do corpo de machos euglossíneos são igualmente úteis e confiáveis para serem utilizadas para a extração e a amplificação do DNA genômico.

Genetic similarity among male bees of Euglossa truncata Rebêlo & Moure (Hymenoptera: Apidae) revealed by RAPD markers.

The genetic RAPD (random amplified polymorphic DNA) markers have been used successfully in taxonomical studies of several groups of organisms. In the present study these molecular markers were used to analyze the genetic similarity among eighteen males of Euglossa truncata Rebêlo & Moure exhibiting variations in two morphological characters (colour of the antennal scape and metaepisternal hairs) which are frequently used to identify this species of euglossine bee. The twelve primers used in the RAPD analysis amplified 127 loci, of which 40 (31.5%) were polymorphic, showing some variation among the individuals. The coefficients of genetic similarity among the individuals ranged from 0.79 to 0.95, indicating a rather high genetic similarity among the 18 male bees studied. No RAPD band was specific to any morphological character analyzed. The results indicate that all bees analyzed belong to the same species. The high genetic similarity among the eighteen euglossine males studied indicates that the variations observed in the morphological characters are not in disagreement with the identification of this species of Euglossina and these characters can vary among males of E. truncata.

Genetic similarity among male bees of Euglossa truncata Rebêlo & Moure (Hymenoptera: Apidae) revealed by RAPD markers

The genetic RAPD (random amplified polymorphic DNA) markers have been used successfully in taxonomical studies of several groups of organisms. In the present study these molecular markers were used to analyze the genetic similarity among eighteen males of Euglossa truncata Rebêlo & Moure exhibiting variations in two morphological characters (colour of the antennal scape and metaepisternal hairs) which are frequently used to identify this species of euglossine bee. The twelve primers used in the RAPD analysis amplified 127 loci, of which 40 (31.5 percent) were polymorphic, showing some variation among the individuals. The coefficients of genetic similarity among the individuals ranged from 0.79 to 0.95, indicating a rather high genetic similarity among the 18 male bees studied. No RAPD band was specific to any morphological character analyzed. The results indicate that all bees analyzed belong to the same species. The high genetic similarity among the eighteen euglossine males studied indicates that the variations observed in the morphological characters are not in disagreement with the identification of this species of Euglossina and these characters can vary among males of E. truncata.

Os marcadores genéticos RAPD (DNA polimórfico amplificado ao acaso) têm sido empregados com sucesso em estudos taxonômicos de diversos grupos de organismos. No presente trabalho esses marcadores moleculares foram utilizados para analisar a similaridade genética entre dezoito machos de Euglossa truncata Rebêlo & Moure apresentando variações em dois caracteres morfológicos (coloração do escapo antenal e da pilosidade dos tufos metaepisternais), os quais normalmente são utilizados na identificação dessa espécie de abelha Euglossina. Os 12 primers utilizados nas análises produziram 127 locos de RAPD, dos quais 40 (31,5 por cento) foram polimórficos, revelando alguma variação genética entre os indivíduos analisados. Os coeficientes de similaridade genética entre os indivíduos variaram de 0,79 a 0,95, indicando alta similaridade entre os 18 machos analisados. Nenhuma banda de RAPD mostrou-se específica para os caracteres morfológicos considerados. Os resultados obtidos indicam que todos os machos analisados pertencem à mesma espécie euglossina. A alta similaridade genética entre os dezoito machos eulossíneos indica que as variações observadas nos caracteres morfológicos destes não estão em discordância com a identificação dessa espécie de Euglossina, e que tais caracteres podem variar entre os machos de E. truncata.

Evaluation of the RAPD profiles from different body parts of Euglossa pleosticta Dressler male bees (Hymenoptera: Apidae, Euglossina).

In the current literature, information is scarce on which part of the adult insect body is suitable for isolation of genomic DNA for genetic analysis based on DNA-markers. In this study, we evaluated RAPD profiles generated from total genomic DNA isolated from distinct body parts (head, legs, thorax + wings and abdomen) of 12 males of Euglossa pleosticta Dressler. From the total of bands analyzed, 9.0% did not show reproducibility. Percent variations of bands in each body segment were: 1.1% (head); 0.4% (legs); 0.8% (thorax/wings) and 6.7% (abdomen). The much higher variation (chi2(one sample) = 10.27; df = 1; P < 0.01) in the RAPD profiles obtained by using DNA isolated from abdomen of the euglossine males suggests that this body part of adult insects should be avoided in DNA extraction procedures. Conversely, the low variation among the RAPD profiles obtained from amplifications of genomic DNA extracted from head, legs and thorax/wings indicates that all these body parts of male bees are equally useful and secure for using in isolation and amplification procedures of total genomic DNA.

Genetic structure analysis of Eufriesea violacea (Hymenoptera, Apidae) populations from southern Brazilian Atlantic rainforest remnants

Random amplified polymorphic DNA (RAPD) markers were used to analyze the genetic structure of Eufriesea violacea populations in three fragments (85.47, 832.58 and 2800 ha) of Atlantic rainforest located in the north of the Brazilian state of Paraná. A total of twelve primers produced 206 loci, of which 129 were polymorphic (95 percent criterion). The proportions of polymorphic loci in each population ranged from 57.28 percent to 59.2 percent, revealing very similar levels of genetic variability in the groups of bees from each fragment. Unbiased genetic distances between groups ranged from 0.0171 to 0.0284, the smallest genetic distance occurring between bees from the two larger fragments. These results suggest that the E. violacea populations from the three fragments have maintained themselves genetically similar to native populations of this species originally present in northern Paraná.