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Genetics and Molecular Biology, Volume: 30, Número: 1, Publicado: 2007
  • CYP1A1 and CYP2E1 polymorphism frequencies in a large Brazilian population Human And Medical Genetics

    Coura, Renata dos Santos; Marques, Christiane de Fátima Silva; Koifman, Rosalina J.; Koifman, Sergio; Cabello, Pedro Hernan; Hatagima, Ana

    Resumo em Inglês:

    The enzymes encoded by the polymorphic genes CYP1A1 and CYP2E1 play an important role in the activation and inactivation of xenobiotics. These enzymes have been associated with xenobiotic-induced diseases, such as cancer, therapeutic failure and adverse effects of drugs. The aim of the present study was to determine the allelic and genotypic frequencies of these polymorphisms in a large, ethnically mixed Brazilian population sample from Rio de Janeiro. Polymorphisms CYP1A1 and CYP2E1 were determined in 870 unrelated individuals by PCR-RFLP analysis in peripheral blood DNA. The observed allelic frequencies were 0.90 for CYP1A1*1A and 0.95 for CYP2E1*1A, in the total sample. The allelic frequency of CYP1A1*2C in "pardos" (0.13) and Brazilian whites (0.11) was higher than in Caucasians (0.05), which may be a result of the Amerindian genetic component, that presents the highest frequency of this allele observed up to now. The genotype distributions for both polymorphisms were in Hardy-Weinberg equilibrium and were statistically different between males and females, and among ethnic groups.
  • Atypical chromosome abnormalities in acute myeloid leukemia type M4 Human And Medical Genetics

    Fett-Conte, Agnes C.; Estrela, Roseli Viscardi; Vendrame-Goloni, Cristina B.; Carvalho-Salles, Andréa B.; Ricci-Júnior, Octávio; Varella-Garcia, Marileila

    Resumo em Inglês:

    This study reports an adult AML-M4 patient with atypical chromosomal aberrations present in all dividing bone marrow cell at diagnosis: t(1;8)(p32.1;q24.2), der(9)t(9;10)(q22;?), and ins(19;9)(p13.3;q22q34) that may have originated transcripts with leukemogenic potential.
  • Molecular characterization of glucose-6-phosphate dehydrogenase deficiency in patients from the southern Brazilian city of Porto Alegre, RS Human And Medical Genetics

    Castro, Simone M. de; Weber, Raquel; Matte, Úrsula; Giugliani, Roberto

    Resumo em Inglês:

    Glucose-6-phosphate dehydrogenase (G6PDH; EC 1.1.1.49) deficiency is one of the most common human enzymopathies throughout the world. Although most affected individuals are asymptomatic, there is a risk of neonatal jaundice and acute hemolytic anemia which can be triggered by infection, some pharmaceuticals and, in older individuals, eating fava beans. We characterized the molecular basis of G6PDH deficiency in a sample of 348 adults from Porto Alegre (population about 1.5 million), the capital of the southernmost Brazilian state of Rio Grande do Sul. Genomic DNA was extracted from peripheral blood leukocytes. We studied the three G6PDH mutations that appear to be the most frequent in Southern Brazil, the G202A and A376G A minus (A-) variants and the C563T Mediterranean (Med) variant. From July 2004 to October 2005, 348 patients (162 Females plus 186 males, age range 0 to 82 years) from Porto Alegre were referred to our laboratory for G6PDH analysis, 36 (9.7%) of which showed deficient G6PDH activity. These 36 patients and 34 randomly-selected non-deficient control individuals were submitted to molecular analysis which revealed a predominance of G6PDH A- allele among the deficient patients. The prevalence of the G6PDH A- variant agrees with its distribution among the ethnic groups that colonized RS, especially those of African, Portuguese, Spanish, and Italian origin.
  • Analysis of the DMPK gene CTG repeat in healthy Brazilians Human And Medical Genetics

    Freitas, Silvia Regina Sampaio; Cabello, Pedro H.; Moura-Neto, Rodrigo Soares; Duro, Luiz Alves

    Resumo em Inglês:

    Myotonic dystrophy (DM) is a neuromuscular disorder caused by the expansion of the cytosine-thymine-guanine (CTG) repeat of the myotonic dystrophy protein kinase gene (DMPK). This repeat is highly polymorphic in healthy individuals [(CTG)5-37], and it has been proposed that expanded CTG alleles originated from larger sized normal alleles [(CTG)19-37]. According to this hypothesis, a positive correlation should be expected between the frequency of these large-sized normal alleles and the prevalence of the disorder in a population. We determined the distribution of CTG alleles of the DMPK gene in 156 healthy Brazilians from Rio de Janeiro city. Our analyses of 312 chromosomes detected 20 different alleles ranging in size from 5 to 27 CTG repeats, with 24 alleles having more than 18 repeats (7.69%). This frequency of (CTG)3(19) alleles observed in our population suggests that the prevalence of DM in Rio de Janeiro should not be different from the prevalence in European populations.
  • Fluorescent in situ hybridization (FISH) as a diagnostic tool for Williams-Beuren syndrome Human And Medical Genetics

    Souza, Deise Helena de; Moretti-Ferreira, Danilo; Rugolo, Lígia Maria Suppo de Souza

    Resumo em Inglês:

    Fluorescent in situ hybridization (FISH) with commercial probes covering the elastin gene (ELN) was used to determine the frequency of the 7q11.23 deletion in 18 children clinically diagnosed with Williams-Beuren syndrome (WBS). A de novo deletion was detected in 15 of the children (83%). Diagnostic investigation for WBS started late in childhood (median = 5.8 years). All the children showed facial features typical of the syndrome, mental retardation and developmental delay. Over-friendliness was observed in the majority of cases. Clinodactyly of the 5th finger (n = 13), cardiovascular disease (n = 9), loquacity (n = 9), low birthweight (n = 8), and failure to thrive (n = 9) were observed only in those children with the deletion. Respiratory problems (n = 9), though not previously reported in the literature, was a common finding in the group studied. Our results confirmed that FISH is useful in identifying 7q11.23 deletions in cases of WBS. Clinical manifestations were more evident in the deletion-positive children.
  • Fluorescence in situ hybridization (FISH) screening for the 22q11.2 deletion in patients with clinical features of velocardiofacial syndrome but without cardiac anomalies Human And Medical Genetics

    Sandrin-Garcia, Paula; Richieri-Costa, Antonio; Tajara, Eloiza Helena; Carvalho-Salles, Andréa Borduchi; Fett-Conte, Agnes Cristina

    Resumo em Inglês:

    The velocardiofacial syndrome (VCFS), a condition associated with 22q11.2 deletions, is characterized by a typical facies, palatal anomalies, learning disabilities, behavioral disturbances and cardiac defects. We investigated the frequency of these chromosomal deletions in 16 individuals with VCFS features who presented no cardiac anomalies, one of the main characteristics of VCFS. Fluorescent in situ hybridization (FISH) with the N25 (D22S75; 22q11.2) probe revealed deletions in ten individuals (62%). Therefore, even in the absence of cardiac anomalies testing for the 22q11.2 microdeletions in individuals showing other clinical features of this syndrome is recommended.
  • Extensive polymorphism and chromosomal characteristics of ribosomal DNA in the characid fish Triportheus venezuelensis (Characiformes, Characidae) Animal Genetics

    Nirchio, Mauro; Oliveira, Claudio; Ferreira, Irani Alves; Granado, Angel; Ron, Ernesto

    Resumo em Inglês:

    The karyotype and chromosomal characteristics of the characid fish Triportheus venezuelensis were investigated using differential staining techniques (C-banding, Ag-NOR staining) and fluorescent in situ hybridization (FISH) with an 18S rDNA probe. The diploid chromosome number (2n = 52), karyotype composition and sex chromosome determination system of the ZZ/ZW type were the same as previously described in other species of the genus Triportheus. However, extensive variation regarding nucleolus organizer regions (NOR) different from other species was observed. 18S rDNA sequences were distributed on nine chromosome pairs, but the number of chromosomes with Ag-NORs was usually lower, reaching a maximum of four chromosomes. When sequential staining experiments were performed, it was demonstrated that: 1.) active NORs usually corresponded to segments with 18S rDNA genes identified in FISH experiments; 2.) several 18S rDNA sequences were not silver-stained, suggesting that they do not correspond to active NORs; and 3.) some chromosomes with silver-stained regions did not display any 18S rDNA signals. These findings characterize an extensive polymorphism associated with the NOR-bearing chromosomes of T. venezuelensis and emphasize the importance of combining traditional and molecular techniques in chromosome studies.
  • Improving the production of transgenic fish germlines: in vivo evaluation of mosaicism in zebrafish (Danio rerio) using a green fluorescent protein (GFP) and growth hormone cDNA transgene co-injection strategy Animal Genetics

    Figueiredo, Márcio de Azevedo; Lanes, Carlos Frederico Ceccon; Almeida, Daniela Volcan; Marins, Luis Fernando

    Resumo em Inglês:

    In fish, microinjection is the method most frequently used for gene transfer. However, due to delayed transgene integration this technique almost invariably produces mosaic individuals and if the gene is not integrated into germ cells its transmission to descendants is difficult or impossible. We evaluated the degree of in vivo mosaicism using a strategy where a reporter transgene is co-injected with a transgene of interest so that potential germline founders can be easily identified. Transgenic zebrafish (Danio rerio) were produced using two transgenes, both comprised of the carp beta-actin promoter driving the expression of either the green fluorescent protein (GFP) reporter gene or the growth hormone cDNA from the marine silverside fish Odonthestes argentinensis. The methodology applied allowed a rapid identification of G0 transgenic fish and also detected which fish were transmitting transgenes to the next generation. This strategy also allowed inferences to be made about genomic transgene integration events in the six lineages produced and allowed the identification of one lineage transmitting both transgenes linked on the same chromosome. These results represent a significant advance in the reduction of the effort invested in producing a stable genetically modified fish lineage.
  • Genetic characterization of the Spanish Trotter horse breed using microsatellite markers Animal Genetics

    Javier Azor, Pedro; Valera, Mercedes; Gómez, María Dolores; Goyache, Félix; Molina, Antonio

    Resumo em Inglês:

    To assist in selection schemes we carried out the first genetic characterization of the Spanish Trotter horse (Trotador Español). We used 16 microsatellite markers to genotype 40 unrelated Spanish trotters, 25 native Balearic horses (11 Menorquina and 14 Mallorquina horses) and 32 Andalusian horses. The observed heterozygosity for the Spanish Trotters was 0.647 ± 0.037 and the expected heterozygosity was 0.696 ± 0.026 while the average number of alleles per locus was 6.0 ± 0.341, these values being similar to the data published for other horse breeds. We also tried to establish the importance of the Mallorquina and Menorquina breeds in the present Spanish Trotter population. Only 9% of the total genetic variability could be attributed to differences between breeds (mean F ST = 0.09 ± 0.010). Recent migration rates were confirmed the low recent genetic relationship between the Balearic breeds and Spanish Trotters, indicating that the genetic background of the present Spanish Trotter population is not based on the native Balearic horse population.
  • Karyological geographic variation of Oligoryzomys nigripes Olfers, 1818 (Rodentia, Cricetidae) from Brazil Animal Genetics

    Paresque, Roberta; Silva, Maria José de Jesus; Yonenaga-Yassuda, Yatiyo; Fagundes, Valéria

    Resumo em Inglês:

    The karyotypes of 85 specimens of Oligoryzomys nigripes (Rodentia, Sigmodontinae) collected in the Cerrado and Atlantic Forest of seven states of Brazil were analyzed. Eighty four specimens presented a karyotype with 2n = 62 and one individual had 2n = 61 due to a monosomy of the X chromosome. High levels of intra- and inter-population karyotypic variability, due to sex chromosomes heteromorphisms and pericentric inversions in four autosomes (pairs 2, 3, 4 and 8), led to a variation of the autosomal arm numbers (fundamental number, FN) from 78 to 82. Synaptonemal complex analyses revealed normal meiosis in males heterozygous for pericentric inversions. We found 39 different cytotypes, 27 of which are herein described for the first time. A literature survey revealed 46 described karyotypes for O. nigripes. We tested the hypothesis that chromosomal variants frequencies are dependent on geographical distribution and we propose a model for the karyotypical evolution of Oligoryzomys nigripes with 2n = 62/FN = 78-82.
  • Chromosome study in Schistocerca (Orthoptera-Acrididae-Cyrtacanthacridinae): karyotypes and distribution patterns of constitutive heterochromatin and nucleolus organizer regions (NORs) Animal Genetics

    Souza, Maria José de; Melo, Natoniel Franklin de

    Resumo em Inglês:

    Chromosome analyses were performed in two grasshopper species of the genus Schistocerca, S. pallens and S. flavofasciata. Both species shared the same diploid number (2n = 23, X in males; 2n = 24, XX in females);and a conserved karyotype composed exclusively of acrocentric chromosomes, but differed in their distribution patterns of constitutive heterochromatin and nucleolus organizer regions (NORs). Constitutive heterochromatin was located in the pericentromeric region of all chromosomes in both species. S. flavofasciata presented an additional C-band on the distal region of the long arm of a small autosome pair (S9). Nucleolus organizer regions (NORs), revealed by silver nitrate staining (Ag-NORs), were observed on a medium autosome pair (M5) in both species. S. pallens presented an additional NOR-bearing autosome (M6). The same sites were labeled after FISH with an rDNA probe in S. pallens cells.
  • Genetic basis of the resistance to Strongyloides venezuelensis (Nematoda, Rhabdiasidae) infection in mice (Mus musculus) Animal Genetics

    Marra, Nelson Mendes; Amarante, Alessandro F.T.; Amarante, Mônica R.V.

    Resumo em Inglês:

    We investigated the resistance to Strongyloides venezuelensis primary infection of mice strains NIH (resistant) and C57BL/6 (susceptible) and the F1 and F2 offspring of crosses between these strains. The mice were infected with 2000 larvae and seven days later were sacrificed for parasite recovery and counting. There was no statistically significant (p > 0.05) sex effect on resistance. The F1 mice showed an intermediate mean number of parasites as compared to the parental NIH and C57BL6 strains. Out of 400 F2 mice, the 10% most resistant mice were infected with 21 to 97 parasites, while the 10% most susceptible mice were infected with 1027 to 1433 parasites. We also found that F2 mice with black fur (n = 72), the same color as the C57BL/6 susceptible parental strain, were more susceptible than white (n = 104) or gray furred (n = 224) mice. It is conceivable that some genes determining coat color are located on the same chromosome as where genes controlling helminth resistance.
  • Karyotype characterization of Crotalaria juncea (L.) by chromosome banding and physical mapping of 18S-5.8S-26S and 5S rRNA gene sites Plant Genetics

    Mondin, Mateus; Santos-Serejo, Janay A.; Aguiar-Perecin, Margarida L.R.

    Resumo em Inglês:

    The chromosomes of Crotalaria juncea, a legume of agronomic interest with a 2n = 16 karyotype composed of metacentric chromosomes, were analyzed using several cytogenetic techniques. C-banding revealed heterochromatic regions around the centromeres in all chromosomes and adjacent to the secondary constriction on the chromosome 1 short arm. Fluorescent staining with the GC-specific chromomycin A3 (CMA) highlighted these heterochromatic regions and a tiny site on the chromosome 1 long arm while the AT-specific stain 4'-6-diamidino-2-phenylindole (DAPI) induced a reversed pattern. Staining with CMA combined with AT-specific distamycin A (DA) counterstaining quenched the pericentromeric regions of all chromosomes, but enhanced fluorescence was observed at the heterochromatic regions around the secondary constriction and on the long arms of chromosomes 1 and 4. Fluorescence in situ hybridization (FISH) revealed 18S-5.8S-26S rRNA gene sites (45S rDNA) on chromosomes 1 and 4, and one 5S rDNA locus on chromosome 1. All the rDNA sites were co-located with the positive-CMA/DA bands, suggesting they were very rich in GC. Silver staining revealed signals at the main 45S rDNA locus on chromosome 1 and, in some cells, chromosome 4 was labeled. Two small nucleoli were detected in a few interphase cells, suggesting that the minor site on chromosome 4 could be active at some stages of the cell cycle.
  • Transference of microsatellite markers from Eucalyptus spp to Acca sellowiana and the successful use of this technique in genetic characterization Plant Genetics

    Santos, Karine Louise dos; Welter, Leocir José; Dantas, Adriana Cibele de Mesquita; Guerra, Miguel Pedro; Ducroquet, Jean Pierre Henri Joseph; Nodari, Rubens Onofre

    Resumo em Inglês:

    The pineapple guava (Acca sellowiana), known in portuguese as the goiabeira-serrana or "Feijoa", is a native fruit tree from southern Brazil and northern Uruguay that has commercial potential due to the quality and unique flavor of its fruits. Knowledge of genetic variability is an important tool in various steps of a breeding program, which can be facilitated by the use of molecular markers. The conservation of repeated sequences among related species permits the transferability of microsatellite markers from Eucalyptus spp. to A. sellowiana for testing. We used primers developed for Eucalyptus to characterize A. sellowiana accessions. Out of 404 primers tested, 180 amplified visible products and 38 were polymorphic. A total of 48 alleles were detected with ten Eucalyptus primer pairs against DNA from 119 A. sellowiana accessions. The mean expected heterozygosity among accessions was 0.64 and the mean observed heterozygosity 0.55. A high level of genetic diversity was also observed in the dendrogram, where the degree of genetic dissimilarity ranged from 0 to 65% among the 119 genotypes tested. This study demonstrates the possibility of transferring microsatellite markers between species of different genera in addition to evaluating the extent of genetic variability among plant accessions.
  • Generation mean analysis for quantitative traits in sesame (Sesamum indicum L.) crosses Plant Genetics

    Sharmila, Vijayarajan; Ganesh, Siruthaiyur Krishnamoorthy; Gunasekaran, Mahalingam

    Resumo em Inglês:

    To study the nature and magnitude of gene effects for yield and its components in sesame (Sesamum indicum L.) we carried out generation mean analysis using the following four crosses of different sesame cultivars: VS 9510 x Co1; NIC 7907 x TMV 3; Cianno 13/10x VRI 1; and Si 1115/1 x TMV 3. The P1, P2, F1, F2, BC1 and BC2 of these generations were studied for seven quantitative traits. The analysis showed the presence of additive, dominance and epistatic gene interactions. The additive dominance model was adequate for plant height in the NIC 7907 x TMV3 and Si 1115/1x TMV 3 crosses and for capsule length in the VS 9510 x Co1, NIC 7907 x TMV 3 and Si 1115/1 x TMV 3 crosses. An epistatic digenic model was assumed for the remaining crosses. Duplicate-type epistasis played a greater role than complementary epistasis. The study revealed the importance of both additive and non-additive types of gene action for all the traits studied.
  • Genetic relationships between Chinese, Japanese, and Brazilian soybean gene pools revealed by simple sequence repeat (SSR) markers Plant Genetics

    Yamanaka, Naoki; Sato, Hiroyuki; Yang, Zhenyu; Xu, Dong He; Catelli, Lizandra Lucy; Binneck, Eliseu; Arias, Carlos Alberto Arrabal; Abdelnoor, Ricardo Vilela; Nepomuceno, Alexandre Lima

    Resumo em Inglês:

    An understanding of the relationship of geographically different soybean gene pools, based on selectively neutral DNA markers would be useful for the selection of divergent parental cultivars for use in breeding. We assessed the relationships of 194 Chinese, 59 Japanese, and 19 Brazilian soybean cultivars (n = 272) using 12 simple sequence repeat (SSR) markers. Quantification Theory III and clustering analyses showed that the Chinese and Japanese cultivars were genetically quite distant to each other but not independent, while Brazilian cultivars were distantly related to the cultivars from the other two countries and formed a cluster that was distant from the other two gene pool clusters. Our results indicated that the Brazilian soybean gene pool is different from the Chinese and Japanese pool. Exchanges of these gene pools might be useful to increase the genetic variability in soybean breeding.
  • Intraspecific differentiation of Chilean isolates of the entomopathogenic fungi Metarhizium anisopliae var. anisopliae as revealed by RAPD, SSR and ITS markers Genetics Of Microorganisms

    Becerra Velásquez, Viviana; Paredes Cárcamo, Mario; Rojo Meriño, Carmen; France Iglesias, Andrés; Franco Durán, Jorge

    Resumo em Inglês:

    The genus Metarhizium consists of a diverse group of asexual entomopathogenic fungi, which have a wide geographical distribution. The Chilean National Agricultural Research Institute (Instituto de Investigaciones Agropecuarias - INIA, Quilamapu Chile) has collected about 350 isolates of Metarhizium anisopliae var. anisopliae from central and southern Chile. These isolates have been partially characterized using morphological traits such as conidia size and shape, colony color, growth pattern and the efficiency of the isolates in controlling specific pests. However, further characterization with molecular markers could detect differences in DNA which could help to better understand the genetic diversity and structure of Chilean populations of this fungus. We analyzed approximately 10% of the INIA collection (39 isolates selected at random) collected from different geographical origins using the polymerase chain reaction (PCR)-random amplified polymorphic DNA (RAPD) method, simple sequences repeat (SSR or microsatellites) analysis and the PCR-restriction fragment length polymorphism (RFLP) assay of internal transcribed spacer (ITS)-rDNA sequences. The RAPD data revealed high genetic diversity in this fungus and an average of 41% of similarity while SSR analysis detected 45.2% similarity and the ITS markers 70.2% similarity. For the three molecular markers, this diversity was not associated with the geographical origin of these isolates.
  • Structural characterization of the bglH gene encoding a beta-glucosidase-like enzyme in an endophytic Bacillus pumilus strain Genetics Of Microorganisms

    Bogas, Andréa C.; Watanabe, Maria Angelica E.; Barbosa, Aneli; Vilas-Boas, Laurival A.; Bonatto, Ana C.; Dekker, Robert; Souza, Emanuel M.; Fungaro, Maria Helena P.

    Resumo em Inglês:

    A beta-glucosidase-like enzyme-encoding gene (bglH) of an endophytic Bacillus pumilus strain (CL16) was cloned using a shotgun genomic library constructed in Escherichia coli. The nucleotide sequence of the entire cloned fragment (2484 bp) was determined and characterized. An incomplete open reading frame (ORF) of 534 bp (ORF1) designated bglP and a complete ORF of 1419 bp (ORF2) designated bglH, located in the fragment, are organized in an operon. The protein deduced from 1419 bp (ORF2) had 472 amino acid residues without a characteristic signal peptide sequence, suggesting that the enzyme is localized in the cytoplasm. The amino acid sequence deduced from bglH gene had high similarity with beta-glucosidases from the glycosyl hydrolase family 1. Over-expression of the B. pumilus bglH gene in E. coli showed a 54 kDa protein whose identity was confirmed by mass spectrometry (MALDI-TOF).
  • Genotoxic and antigenotoxic effects of Fucus vesiculosus extract on cultured human lymphocytes using the chromosome aberration and Comet assays Mutagenesis

    Leite-Silva, Cleide; Gusmão, Cássia Lima Silva; Takahashi, Catarina Satie

    Resumo em Inglês:

    The brown seaweed Fucus vesiculosus (Fucales, Fucaceae) was screened for its protective activity using doxorubicin-induced DNA damage in human lymphocytes. In this study, we assessed the genotoxic and antigenotoxic potential of three different concentrations (0.25, 0.5 and 1.0 mg mL-1) of F. vesiculosus aqueous extract using the chromosome aberration and Comet assays. Treatment of human lymphocyte cultures with 0.25, 0.5 and 1.0 mg mL-1 F. vesiculosus aqueous extract had no effect on the chromosome aberration frequency or on the extent of DNA damage detected by the Comet assay. The antigenotoxic effects of the extract were tested in human lymphocyte cultures treated with 15 µg mL-1 of doxorubicin, either alone or combined with the different concentrations of the extract, which was added to the cultures before, simultaneously with or after the doxorubicin. Only when lymphocytes were pre-treated with extract there was a reduction in doxorubicin-induced chromosome aberrations and DNA damage as detected by the Comet assay. These results demonstrate that F. vesiculosus aqueous extract is not genotoxic in cultured human lymphocytes and indicate that when added to lymphocyte cultures before doxorubicin it has antigenotoxic activity against doxorubicin-induced DNA damage.
  • Evidence of high inbreeding in a population of the endangered giant anteater, Myrmecophaga tridactyla (Myrmecophagidae), from Emas National Park, Brazil Evolutionary Genetics

    Collevatti, Rosane G.; Leite, Kelly C.E.; Miranda, Guilherme H.B. de; Rodrigues, Flavio H.G.

    Resumo em Inglês:

    We report the genetic structure, relatedness and mating structure of a population of the endangered giant anteater Myrmecophaga tridactyla Linnaeus, 1758 in the Emas National Park, Brazil, based on variability at five microsatellite loci. Additionally, we addressed the hypothesis that the M. tridactyla population studied has low levels of polymorphism and high levels of inbreeding and relatedness and that animals with overlapping home range are highly related. All five microsatellite loci displayed low levels of polymorphism and of expected and observed heterozygosity. The low level of polymorphism and high inbreeding showed by the population studied may be the outcome of high mortality and reduction in population size due to recurrent fire events in the Emas National Park, as reported in 1994. The reduction in population size may have led to a higher frequency of mating between closely related animals, augmented by the isolation of the population in the park because of the expansion of agricultural land and fragmentation of the Cerrado environment. The natural history of M. tridactyla and the phylopatric (sex-biased dispersal) behavior of females should increase the effects of isolation and bottlenecking, decreasing gene flow and increasing inbreeding. However, the low levels of polymorphism found in this population may simply be due to the natural history and evolution of M. tridactyla as reported for other species. The genetic structure and dynamics of this population needs to be investigated more profoundly in order to provide sound data for the design of conservation strategies for M. tridactyla in the Emas National Park.
  • Extinction of canid populations by inbreeding depression under stochastic environments in Southwestern Goiás State: a simulation study Evolutionary Genetics

    Rodrigues, Flávia Melo; Diniz-Filho, José Alexandre Felizola

    Resumo em Inglês:

    A frequently addressed question in conservation biology is what is the chance of survival for a population for a given number of years under certain conditions of habitat loss and human activities. This can be estimated through an integrated analysis of genetic, demographic and landscape processes, which allows the prediction of more realistic and precise models of population persistence. In this study, we modeled extinction in stochastic environments under inbreeding depression for two canid species, the maned wolf (Chrysocyon brachiurus) and the crab-eating fox (Cerdocyon thous), in southwest Goiás State. Genetic parameters were obtained from six microsattelite loci (Short Tandem Repeats - STR), which allowed estimates of inbreeding levels and of the effective population size under a stepwise mutation model based on heterozygosis. The simulations included twelve alternative scenarios with varying rates of habitat loss, magnitude of population fluctuation and initial inbreeding levels. ANOVA analyses of the simulation results showed that times to extinction were better explained by demographic parameters. Times to extinction ranged from 352 to 844, in the worst and best scenario, respectively, for the large-bodied maned wolf. For the small-bodied crab-eating fox, these same estimates were 422 and 974 years. Simulations results are within the expectation based on knowledge about species' life history, genetics and demography. They suggest that populations can persist through a reasonable time (i.e., more than 200 years) even under the worst demographic scenario. Our analyses are a starting point for a more focused evaluation of persistence in these populations. Our results can be used in future research aiming at obtaining better estimates of parameters that may, in turn, be used to achieve more appropriate and realist population viability models at a regional scale.
  • Phylogenetic position of Placozoa based on large subunit (LSU) and small subunit (SSU) rRNA genes Evolutionary Genetics

    Silva, Fernanda Britto da; Muschner, Valéria C.; Bonatto, Sandro L.

    Resumo em Inglês:

    The phylum Placozoa, represented by its single species Trichoplax adhaerens, has always been considered one of the most basal animal clades due to its low morphological complexity. However, despite its importance, the phylogenetic position of the Placozoa remains uncertain, as does the relationships between the basal metazoans. We sequenced the nearly complete large subunit (LSU) of the T. adhaerens rRNA gene together with the small subunit (SSU) rRNA and used several methods to infer the phylogenetic position of Placozoa and the relationships between the basal animal phyla. The phylogenetic trees obtained from the LSU, SSU and SSU + LSU datasets support monophyly of all basal metazoan phyla, except the Porifera. However, the relationships between all these phyla were not consistently resolved due to a strong conflict between the LSU and SSU phylogenetic signals. The hypotheses that Placozoa is a derived Cnidaria or the most basal metazoan were not supported by our data. Moreover, our analyses suggests that Placozoa is most likely the sister group of Cnidaria and/or Bilateria.
  • Distribution and conservation of the transposable element gypsy in drosophilid species Evolutionary Genetics

    Herédia, Fabiana; Loreto, Elgion Lúcio S.; Valente, Vera Lúcia S.

    Resumo em Inglês:

    In an attempt to understand the dynamics of transposable elements (T'S) in the genome of host species, we investigated the distribution, representativeness and conservation of DNA sequences homologous to the Drosophila melanogaster gypsy retrotransposon in 42 drosophilid species. Our results extended the knowledge about the wide distribution of gypsy in the genus Drosophila, including several Neotropical species not previously studied. The gypsy-like sequences showed high divergence compared to the D. melanogaster gypsy element. Furthermore, the conservation of the restriction sites between gypsy sequences from phylogenetically unrelated species pointed to a more complex evolutionary picture, which includes the possibility of the horizontal transfer events already described for this retrotransposon.
  • ITS2 variability of Biomphalaria (Mollusca, Planorbidae) species from the Paranapanema Valley (São Paulo State, Brazil): diversity patterns, population structure, and phylogenetic relationships Evolutionary Genetics

    Tuan, Roseli; Santos, Priscila dos

    Resumo em Inglês:

    The ribosomal DNA internal transcribed spacer 2 (ITS2) has been shown to be a useful genetic marker for species identification and phylogenetic reconstruction in the genus of freshwater snails Biomphalaria (Preston 1910). Additionally, ITS2 studies in Biomphalaria have uncovered significant intra-specific genetic variability suggesting the presence of cryptic species complexes. We obtained ITS2 sequences for the Biomphalaria species B. glabrata, B. tenagophila, B. occidentalis and B. peregrina from the Paranapanema Valley (São Paulo State, Brazil) and compared them with a comprehensive set of published Biomphalaria ITS2 sequences using Bayesian inference of phylogeny. Analysis of the resulting trees showed that the newly obtained B. glabrata sequences did not cluster with those from other Brazilian localities and that sub-structuring occurred among Brazilian B. tenagophila populations. Moreover, although ITS2 sequences seem to indicate clear genetic differentiation within both B. glabrata and B. tenagophila, evidence in support of the occurrence of cryptic species is more compelling for the latter. We discuss the significance and implications of the detected patterns of ITS2 variability for taxonomic studies in Biomphalaria.
  • Asymmetry of frontal bristles and postocular setae in species and hybrids of the Anastrepha fraterculus complex (Diptera, Tephritidae) Developmental Genetics

    Souza, João Maria G.A.; Gouveia, Milson; Perondini, André L.P.; Selivon, Denise

    Resumo em Inglês:

    Asymmetry of the frontal bristles and postocular setae was studied in samples from natural populations and laboratory colonies of Anastrepha sp. 1 aff. fraterculus, of A. sp. 2 aff. fraterculus, and in F1 hybrids obtained from laboratory reciprocal crosses. Natural populations were sampled in a zone of sympatry and in two geographically distant regions with different climatic conditions. Asymmetry was scored as the differences between the number of bristles and of setae on the right and left sides of the head, males and females analyzed independently. The two traits exhibited variability according to the model of fluctuating asymmetry (FA). No significant differences among samples were found in the FA of frontal bristles. A significant FA was observed for the postocular setae of A. sp. 1 males from a southern population (Vacaria, RS) as compared to the asymmetry exhibited by males and females of some other samples. No significant differences in FA were observed among the interspecific hybrids and the laboratory samples of both parental species. The higher FA found in the males from Vacaria was attributed to climatic conditions prevailing in that region. The absence of a higher FA in hybrids may be related to the relatively recent evolutionary history of the two species.
  • Temperature and the progeny sex-ratio in Sciara ocellaris (Diptera, Sciaridae) Developmental Genetics

    Nigro, Rogério G.; Campos, Maria Cristina C.; Perondini, André Luiz P.

    Resumo em Inglês:

    We found that the sex-ratio of an amphigenic strain of Sciara ocellaris varied widely from progenies with few males to progenies containing a larger proportion of males, with single-sex progenies being rare. The sex-ratio distributions were dependent on the temperature at which the stocks of flies were raised, with the sex-ratio distributions being symmetrical (i.e. about 50% males) at 18 °C and 20 °C while at the higher temperatures of 24 °C and 28 °C the distributions were skewed toward a high proportion of females with the mean proportion of males decreasing to about 30-37% per progeny. Temperature-shift experiments showed that high temperatures were effective only during the last stages of female pupal development plus a period after adult emergence, stages corresponding to oocyte maturation. When imagine females were exposed to temperatures as low as 12 °C the sex-ratio distributions of their progeny were skewed toward a high proportion of males per progeny. No differential fecundity was involved in these progeny sex-ratio modifications. Egg-to-adult survival was lower at 18 °C and 28 °C but no correlations with skewing in the sex ratio distributions were observed, indicating that modifications in progeny sex-ratio did not involve the differential survival of a particular sex.
  • Developmental and hyperthermia-induced expression of the heat shock proteins HSP60 and HSP70 in tissues of the housefly Musca domestica: an in vitro study Developmental Genetics

    Sharma, Sunita; Rohilla, Manoj Singh; Tiwari, Pramod Kumar

    Resumo em Inglês:

    The expression pattern of two major chaperones, the heat shock proteins (HSPs) HSP60 and HSP70 was studied in vitro in tissues of the housefly Musca domestica during larval and adult stages of development to identify their immunological relatives and understand their functional significance in normal cellular activities and during thermal stress. Fluorographs of labeled polypeptides and western blots demonstrated that both HSPs are expressed constitutively and heat-induced in all the larval and adult cell types examined. The pattern of whole tissue immunocytochemical staining using anti-HSP60 and anti-HSP70 antibodies corresponded well with the observations from western blots or fluorographs. In developing oocytes, both constitutive and heat inducible expression of HSP60 were regulated in an oocyte stage-specific manner. In unstressed ovaries the expression of these proteins was less pronounced in early stage oocytes (1st - 8th) than at later stages (9th and onward). The heat shock, however, induced both HSP70 and HSP60 to a significantly high level in early stage oocytes (1st-8th) as compared to their respective controls. Our findings indicate the involvement of the HSP60 and HSP70 proteins in the development, growth and differentiation of both somatic and germ line tissues. Furthermore, the enhanced co-expression of HSP70 and HSP60 upon heat shock in various larval and adult cell types suggests the possible role of HSP60 in thermoprotection.
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