Between the cross and the sword: the crisis of the gene concept El-Hani, Charbel Niño Abstract in English: Challenges to the gene concept have shown the difficulty of preserving the classical molecular concept, according to which a gene is a stretch of DNA encoding a functional product (polypeptide or RNA). The main difficulties are related to the overlaying of the Mendelian idea of the gene as a ‘unit’: the interpretation of genes as structural and/or functional units in the genome is challenged by evidence showing the complexity and diversity of genomic organization. This paper discusses the difficulties faced by the classical molecular concept and addresses alternatives to it. Among the alternatives, it considers distinctions between different gene concepts, such as that between the ‘molecular’ and the ‘evolutionary’ gene, or between ‘gene-P’ (the gene as determinant of phenotypic differences) and ‘gene-D’ (the gene as developmental resource). It also addresses the process molecular gene concept, according to which genes are understood as the whole molecular process underlying the capacity to express a particular product, rather than as entities in ‘bare’ DNA; a treatment of genes as sets of domains (exons, introns, promoters, enhancers, etc.) in DNA; and a systemic understanding of genes as combinations of nucleic acid sequences corresponding to a product specified or demarcated by the cellular system. In all these cases, possible contributions to the advancement of our understanding of the architecture and dynamics of the genetic material are emphasized. |
Human and Medical Genetics Article Molecular analysis of three FUT3 gene single nucleotide polymorphisms and their relationship with the lewis erythrocytary phenotype in a human population of japanese-ancestry living in Tomé Açu, a town in the Brazilian Amazon Francez, Pablo Abdon da Costa Corvelo, Tereza Cristina de Oliveira Silva, Flávio Ricardo Leal da Santos, Sidney Emanuel Batista dos Abstract in English: The Lewis blood group system involves two major antigens, Leª and Le b. Their antigenic determinants are not primary gene products but are synthesized by the transfer of sugar subunits to a precursory chain by a specific enzyme which is the product of the FUT3 gene (Lewis gene). The presence of three FUT3 gene single nucleotide polymorphisms (SNPs) (59T > G; 508G > A and 1067T > A) was related to the Lewis phenotype of erythrocytes from 185 individuals of Japanese ancestry living in the town of Tomé-Açu in the Brazilian Amazon region. This relationship was detected using a serological hemagglutination test and the Dot-ELISA assay along with the molecular technique PCR-RFLP. We found that the three SNPs investigated in this study only accounted for a proportion of the Lewis-negative phenotype of the erythrocytes. |
Human and Medical Genetics Programmed cell death 1 gene (PDCD1) polymorphism and pemphigus foliaceus (fogo selvagem) disease susceptibility Braun-Prado, Karin Petzl-Erler, Maria Luiza Abstract in English: Pemphigus foliaceus, also known as fogo selvagem, is an autoimmune disease of the epidermis characterized by superficial blisters and antibodies against desmoglein 1. It is a multifactorial disease and genetic susceptibility is oligogenic or polygenic. Considering the crucial function of the programmed cell death 1 molecule (PD-1) in the immune response, the aim of this study was to verify if variants of the PDCD1 gene influence susceptibility and resistance to pemphigus foliaceus, in a case - control disease association study. We analyzed patients (n = 154) and unaffected control individuals (n = 325) of the Brazilian population, in respect to the PD1.3(G,A) PD1.5(C,T) and PD1.6(A,G) single nucleotide polymorphisms (SNPs) and also investigated, for the first time, the exon 5 PDCD1 microsatellite (CTG)n. The patient and control samples were divided into strata, according to the predominant ancestry of the individuals (African or European). The PD1.5 genotype distribution in the patients sample was almost indistinguishable from that in the control sample, in both population strata. A possible negative association between pemphigus foliaceus and allele PD1.3A was observed in the total African and European ancestry population sample (odds ratio (OR) = 0.55, p = 0.066) and should be investigated in forthcoming studies. The PD1.6A allele was over-represented among the patients of predominantly European ancestry due to an increase of both the G/A and the A/A genotypes (OR = 2.12 and 1.74, respectively; p = 0.035). We conclude that polymorphisms of the PDCD1 gene may influence susceptibility to pemphigus foliaceus, at least in Brazilians of predominantly European ancestry. |
Human and Medical Genetics Evaluation of gene expression profiling in a mouse model of L-gulonolactone oxidase gene deficiency Yan, Jian Jiao, Yan Li, Xinmin Jiao, Feng Beamer, Wesley G. Rosen, Cliff J. Gu, Weikuan Abstract in English: Humans and guinea pigs are species which are unable to synthesize ascorbic acid (vitamin C) because, unlike rodents, they lack the enzyme L-gulonolactone oxidase (Gulo). Although the phenotype of lacking vitamin C in humans, named scurvy, has long been well known, information on the impact of lacking Gulo on the gene expression profiles of different tissues is still missing. This knowledge could improve our understanding of molecular pathways in which Gulo may be involved. Recently, we discovered a deletion that includes all 12 exons in the gene for Gulo in the sfx mouse, characterized by spontaneous bone fractures. We report here the initial analysis of the impact of the Gulo gene deletion on the murine gene expression profiles in the liver, femur and kidney. |
Human and Medical Genetics Preimplantation genetic diagnosis of Von Hippel-Lindau disease cancer syndrome by combined mutation and segregation analysis Sumita, Denilce R. Rocha, José Cláudio C. Iaconelli Jr., Assumpto Borges Jr., Edson Pereira, Lygia V. Abstract in English: Von Hippel-Lindau (VHL) disease is an autosomal dominant cancer syndrome, associated with the development of tumors and cysts in multiple organ systems, whose expression and age of onset are highly variable. The VHL disease tumor suppressor gene (VHL) maps to 3p25-p26 and mutations ranging from a single base change to large deletions have been detected in patients with VHL disease. We developed a single cell PCR protocol for preimplantation genetic diagnosis (PGD) of VHL disease to select unaffected embryos on the basis of the detection of the specific mutation and segregation analysis of polymorphic linked markers. Multiplex-nested PCR using single buccal cells of an affected individual were performed in order to test the accuracy and reliability of this single-cell protocol. For each locus tested, amplification efficiency was 83% to 87% and allelic drop-out rates ranged from 12% to 8%. Three VHL disease PGD cycles were performed on cells from a couple with paternal transmission of a 436delC mutation in exon 2 of the VHL gene, leading to the identification of three unaffected embryos. Independent of the mutation present, this general PGD protocol for the diagnosis of VHL disease can be used in families informative for either the D3S1038 or D3S1317 microsatellite markers. |
Human and Medical Genetics JAK2 V617F prevalence in Brazilian patients with polycythemia vera, idiopathic myelofibrosis and essential thrombocythemia Monte-Mór, Bárbara da Costa Reis Cunha, Anderson Ferreira da Pagnano, Kátia Bórgia Barbosa Saad, Sara Terezinha Lorand-Metze, Irene Costa, Fernando Ferreira Abstract in English: Polycythemia vera (PV), essential thrombocythemia (ET) and idiopathic myelofibrosis (IMF) are myeloproliferative disorders (MPD) that arise from the clonal proliferation of a pluripotent hematopoietic progenitor, leading to the overproduction of one or more myeloid lineages. Recently, a specific mutation in the JAK2 gene, which encodes a tyrosine kinase, has been shown to be associated with the myeloproliferative phenotype observed in PV, ET and IMF. In this study of Brazilian patients, the JAK2 V617F mutation [c.1887G > T) was detected in four out of 49 patients with PV (96%), 14 out of 25 patients with IMF (56%), and in eight out of 29 patients with ET, which is in accordance with previous screenings of this mutation in other populations. |
Human and Medical Genetics Widening the clinical spectrum of Pitt-Rogers-Danks/Wolf-Hirschhorn syndromes Mazzeu, Juliana F. Krepischi-Santos, Ana Cristina Rosenberg, Carla Lourenço, Charles M. Lezirovitz, Karina Szuhai, Karoly Martelli, Lúcia R. Vianna-Morgante, Angela M. Abstract in English: Chromosomal rearrangements involving partial deletion of the short arm of chromosome 4 and partial duplication of the short arm of chromosome 8 have been described both in Pitt-Rogers-Danks syndrome (PRDS) and Wolf-Hirschhorn syndrome (WHS), the former being considered a milder phenotype of the latter. We describe a patient with partial deletion of chromosome 4 and partial duplication of chromosome 8 documented by array-comparative genomic hybridization (Array-CGH). In addition to the typical features of PRDS, the patient exhibited some clinical signs (genital hypoplasia, radioulnar synostosis and mesomelic limb shortness) infrequently, or never previously, reported in PRDS. These findings broaden the spectrum of anomalies generally associated with these syndromes. |
Animal Genetics Bayesian inference on field data for genetic parameters for some reproductive and related traits of Nellore cattle (Bos indicus) Faria, Carina Ubirajara de Magnabosco, Cláudio de Ulhôa Reyes, Arcadio de los Lôbo, Raysildo Barbosa Bezerra, Luiz Antônio Framartino Sainz, Roberto Daniel Abstract in English: We used Gibbs sampling in single and two-trait animal models to estimate genetic parameters for some reproductive and related traits of Nellore cattle (Bos indicus). Female traits were age at first calving (AFC), cumulative productivity (CP) and adult weight (AW). For males, scrotal circumferences at 365 and 450 days of age were analyzed. Gibbs sampling using the ‘Multiple Trait using Gibbs Sampling under Animal Model’ (MTGSAM) program of Van Tassel and Van Vleck was used to estimate the (co)variance components of the traits and conduct genetic analyses. Heritabilities were AFC = 0.26, AW = 0.36 and CP = 0.25 under the single-trait animal model. The mean, mode and median estimates for genetic parameters obtained from the marginal posterior distributions were similar for all traits except AW, which presented lower values for the mode than the mean and median. However, the marginal posterior distributions for the traits studied presented a tendency toward normality. Favorable, but low, negative genetic correlations were found between male scrotal circumference and female age at first calving. The reproductive traits showed medium-magnitude heritabilities, which indicates that these traits should respond to selection and therefore should be included in genetic improvement programs. |
Animal Genetics Genetic evaluation for persistency of lactation in Holstein cows using a random regression model Cobuci, Jaime Araujo Euclydes, Ricardo Frederico Costa, Claudio Napolis Torres, Robledo de Almeida Lopes, Paulo Sávio Pereira, Carmen Silva Abstract in English: A model for analyzing test day records including both fixed and random coefficients was applied to the genetic evaluation of first lactation data for Holstein cows. Data comprising 87045 test-day milk yield records from calving between 1997 and 2001 from Holstein herds in 10 regions of the Brazilian state of Minas Gerais. Six persistency of lactation measures were evaluated using breeding values obtained by random regression analyses. The Wilmink function was used to model the additive genetic and permanent environmental effects. Residual variance was constant throughout lactation. Ranking for animals did not change among criteria for persistency measurements, but ranking changes were observed when the estimated breeding value (EBV) for persistency of lactation was contrasted with those estimated for 305-day milk yield (305MY). The rank correlation estimates for persistency of lactation and 305MY were practically the same for sire and cows, and ranged from -0.45 to 0.69. The EBVs for milk yield during lactation for sires producing daughters with superior 305MY indicate genetic differences between sires regarding their ability to transmit desirable persistency of lactation traits. This suggests that selection for total lactation milk yield does not identify sires or cows that are genetically superior in regard to persistency of lactation. Genetic evaluation for persistency of lactation is important for improving the efficiency of the milk production capacity of Holstein cows. |
Animal Genetics Structure and genetic relationships between Brazilian naturalized and exotic purebred goat domestic goat (Capra hircus) breeds based on microsatellites Oliveira, Joelliton Domingos de Igarashi, Maria Luiza Silveira de Paiva Machado, Théa Mírian Medeiros Miretti, Marcos Mateo Ferro, Jesus Aparecido Contel, Eucleia Primo Betioli Abstract in English: The genetic relationships and structure of fourteen goat (Capra hircus) populations were estimated based on genotyping data from 14 goat populations (n = 410 goats) at 13 microsatellite loci. We used analysis of molecular variance (AMOVA), principal component analysis (PCA) and F statistics (F IS, F IT and F ST) to evaluate the genetic diversity (Ho, He and ad) of the goats. Genetic distances between the 14 goat populations were calculated from allelic frequency data for the 13 microsatellite markers. Moderate differentiation was observed for the populations of the undefined breeds (including the Anglo-Nubian-M breed), the naturalized Brazilian breeds (Moxotó, Canindé), the exotic purebred breeds (Alpine, Saanen, Toggenbourg and Anglo-Nubian) and the naturalized Brazilian Graúna group. Our AMOVA showed that a major portion (88.51%) of the total genetic variation resulted from differences between individual goats within populations, while between-populations variation accounted for the remaining 11.49% of genetic variation. We used a Reynolds genetic distance matrix and PCA to produce a phenogram based on the 14 goat populations and found three clusters, or groups, consisting of the goats belonging to the undefined breed, the naturalized breeds and the exotic purebred breeds. The closer proximity of the Canindé breed from the Brazilian state of Paraíba to the Graúna breed from the same state than to the genetically conserved Canindé breed from the Brazilian state of Ceará, as well as the heterozygosity values and significant deviations from Hardy-Weinberg equilibrium suggests that there was a high number of homozygotes in the populations studied, and indicates the importance of the State for the conservation of the local breeds. Cataloguing the genetic profile of Brazilian goat populations provides essential information for conservation and genetic improvements programs. |
Animal Genetics Quantitative trait loci (QTL) mapping for growth traits on bovine chromosome 14 Miyata, Marcelo Gasparin, Gustavo Coutinho, Luiz Lehmann Martinez, Mario Luiz Machado, Marco Antonio Silva, Marcos Vinicius G. Barbosa da Campos, Ana Lucia Sonstegard, Tad S. Rosário, Millor Fernandes do Regitano, Luciana Correia de Almeida Abstract in English: Quantitative trait loci (QTL) mapping in livestock allows the identification of genes that determine the genetic variation affecting traits of economic interest. We analyzed the birth weight and weight at 60 days QTL segregating on bovine chromosome BTA14 in a F2 resource population using genotypes produced from seven microsatellite markers. Phenotypes were derived from 346 F2 progeny produced from crossing Bos indicus Gyr x Holstein Bos taurus F1 parents. Interval analysis to detect QTL for birth weight revealed the presence of a QTL (p < 0.05) at 1 centimorgan (cM) from the centromere with an additive effect of 1.210 ± 0.438 kg. Interval analysis for weight at 60 days revealed the presence of a QTL (p < 0.05) at 0 cM from the centromere with an additive effect of 2.122 ± 0.735 kg. The region to which the QTL were assigned is described in the literature as responsible for some growth traits, milk yield, milk composition, fat deposition and has also been related to reproductive traits such as daughter pregnancy rate and ovulation rate. The effects of the QTL described on other traits were not investigated. |
Animal Genetics Associations of MYF5 gene polymorphisms with meat quality traits in different domestic pig (Sus scrofa) populations Liu, Min Peng, Jian Xu, Dequan Zheng, Rong Li, Feng’e Li, Jialian Zuo, Bo Lei, Minggang Xiong, Yuanzhu Deng, Changyan Jiang, Siwen Abstract in English: The MYF5 gene is first inducibly expressed in muscle cell during embryonic muscle development and plays an important role in regulating the differentiation of skeletal muscle precursors. In this study we used PCR-RFLP to investigate two pig (Sus scrofa) populations (n = 302) for two MYF5 gene polymorphisms, a previously unreported novel Met-Leu shift single nucleotide polymorphism (SNP) MYF5/Hsp92II located on exon 1 and the previously identified intron 1 MYF5/HinfI SNP. Haplotype and association analysis showed that haplotypes of the two SNPs were significantly associated with drip loss rate (DLR, p < 0.05), water holding capacity (WHC, p < 0.05), biceps femoris meat color value (MCV2, p < 0.05), biceps femoris marbling score (MM2, p < 0.01), longissimus dorsi intramuscular fat percentage (IMF, p < 0.01) and longissimus dorsi Water moisture content (WM, p < 0.01) in the population 2. However, further studies are needed to confirm these preliminary results. |
Animal Genetics Comparative karyotype analysis in diploid and triploid Dolichoplana carvalhoi (Tricladida, Terricola, Rhynchodemidae) from Brazil Alvarez, Luciana Almeida, Eunice Judith Cardoso de Abstract in English: In this work, we present cytogenetic data for the land planarian Dolichoplana carvalhoi. Two different karyotypes, one diploid (2n = 2x = 14 chromosomes) and one triploid (2n = 3x = 21 chromosomes), corresponding to two morphological body patterns, are described. Chromosomes from regenerating blastema were studied after routine Giemsa staining and CBG banding. Our analyses revealed heteromorphisms in chromosomes 2, 3 and 4 of the diploid karyotype and in chromosomes 1, 3 and 7 of the triploid karyotype. Further studies are needed in order to determine if the two morphological patterns of D. carvalhoi represent distinct species. |
Animal Genetics Application of a double-enrichment procedure for microsatellite isolation and the use of tailed primers for high throughput genotyping Diniz, Fábio Mendonça Iyengar, Arati Lima, Paulo Sarmanho da Costa Maclean, Norman Bentzen, Paul Abstract in English: The number of microsatellite loci and their allelic diversity contribute to increase accuracy and informativity of genetic estimates, however, the isolation of microsatellite loci is not only laborious but also quite expensive. We used (GATA)n and (GACA)n tetranucleotide probes and single- and double-enrichment hybridization to construct and screen a genomic library with an increased proportion of DNA fragments containing repeat motifs. Repeats were found using both types of hybridization but the double-enrichment procedure recovered sequences of which 100% contained (GATA)n and (GACA)n motifs. Microsatellite loci primers were then designed with an M13R-tail or CAG-tag to produce scorable PCR products with minimal stutter. The approach used in this study suggests that double-enrichment is a worthwhile strategy when isolating repeat motifs from eukaryotic genomes. Moreover, the use of tailed microsatellite primers provides increased resolution for compound microsatellite loci, with a significant decrease in costs. |
Plant Genetics A novel statistical method for assessing SSR variation in autotetraploid alfalfa (Medicago sativa L.) Liu, Zhi-Peng Liu, Gong-She Yang, Qing-Chuan Abstract in English: The level of variation of simple sequence repeat (SSR) markers in cultivated alfalfa from American, Australian and Chinese sources was evaluated using a novel autotetraploid statistical method to calculate the effective number of alleles, the allele frequencies and heterozygosity. We used 19 SSR primers to screen seven polymorphic SSR loci in 320 plants from eight populations. The genetic distance and phylogenetic analysis (DISPAN) program was used to calculate the inter- and intra-population genetic relationships using the conventional binary absence/presence (0/1) method and our novel autotetraploid method. The autotetraploid method resulted in significantly higher heterozygosity (p < 0.01), average effective number (p < 0.01) and lower standard genetic distance (p < 0.01) than the binary method. Our results suggest that our new autotetraploid method is a very useful tool for assessing genetic variation and genetic relationships in all autotetraploid plant species. |
Plant Genetics Association between genetic distances in wheat (Triticum aestivum L.) as estimated by AFLP and morphological markers Vieira, Eduardo Alano Carvalho, Fernando Irajá Félix de Bertan, Ivandro Kopp, Mauricio Marini Zimmer, Paulo Dejalma Benin, Giovani Silva, José Antônio Gonzalez da Hartwig, Irineu Malone, Gaspar Oliveira, Antônio Costa de Abstract in English: Molecular and morphological data analyses matrices are very informative tools for the estimation of genetic distances. We used AFLP markers, morphological traits and combined analysis to estimate the genetic distances between wheat genotypes and ascertain any associations between the two techneques. Nineteen wheat (Triticum aestivum L.) genotypes were analyzed using amplified fragment length polymorphism (AFLP) markers and field evaluated for two years. The matrices obtained by morphological and molecular marker data analyses revealed a significant but moderate correlation (r = 0.47), indicating that such techniques sample distinct genome regions. The combined analysis was found to be highly correlated with AFLP (r = 0.97) and moderately correlated with morphological (r = 0.59) markers. A possible explanation for such results is a bias caused by the much higher number of AFLP (229) than morphological (17) markers. Thus, it is evident that the combined analysis is not efficient when a very dissimilar number of markers are used in each isolated technique. Therefore, to obtain a better knowledge of the degree of divergence among genotypes it is necessary to consider each analysis separately. |
Plant Genetics Genetic structure of Brazilian wild rice (Oryza glumaepatula Steud., Poaceae) populations analyzed using microsatellite markers Karasawa, Marines M.G. Vencovsky, Roland Silva, Cynthia M. Zucchi, Maria Imaculada Oliveira, Giancarlo C.X. Veasey, Elizabeth A. Abstract in English: Knowledge of the genetic structure and diversity of natural populations is important in developing strategies for in situ and ex situ conservation. We used eight microsatellite loci to estimate genetic structure and investigate within and between population genetic variation in eleven Brazilian wild rice (Oryza glumaepatula) populations. The study showed the following genetic diversity parameters: average number of 3.1 alleles per locus; 77.3% polymorphic loci; 0.091 observed heterozygosity and 0.393 gene diversity. F-statistics detected by microsatellite loci were: F ST = 0.491 (and R ST = 0.608), F IS = 0.780 and F IT = 0.888. No population was in Hardy-Weinberg equilibrium. The estimated apparent outcrossing rate (0.143) indicated a predominance of self-fertilization. The gene flow values were low (Nm = 0.259 and 0.161 for F ST and R ST, respectively). Populations were spatially structured but without a correlation between genetic and geographic distances. Five populations (PG-4, PG-2, PU-1, SO-4, NE-18) were identified as priorities for conservation strategies. Populations from the Amazon biome showed heterogeneity with respect to intrapopulation diversity. The high level of genetic differentiation between populations and the high number of private alleles suggested that sampling should be carried out on a large number of O. glumaepatula populations for ex situ conservation purposes. |
Plant Genetics Combining ability analysis for grain filling duration and yield traits in spring wheat (Triticum aestivum L. em. Thell.) Kamaluddin, Singh, Rishi M. Prasad, Lal C. Abdin, Malik Z. Joshi, Arun K. Abstract in English: A diallel analysis of wheat (Triticum aestivum L. em. Thell) parents (n = 11) and their F1 (n = 55) and F2 (n = 55) offspring was carried out for the following four traits: grain filling duration (GFD), GFD for growing degree days (GDD), 1000 seed weight and seed yield per plant. Analysis of variance for general combining ability (GCA) and specific combining ability (SCA) displayed significant F1 and F2 general and specific combining ability effects for the four traits studied. For all the traits the GCA effects were relatively more important than the SCA effects, indicating that additive genetic effects were predominant. Crosses displaying high SCA effects for grain filling duration, seed weight and yield were observed to be derived from parents having various types of GCA effects (high x high, high x low, low x low and medium x low). The single seed descent method can be applied to exploit additive gene effects whereas dominance gene effects could be valuable in hybrid wheat breeding programs. Among the parents, genotypes from the International Maize and Wheat Improvement Center (Centro Internacional de Mejoramiento de Maíz y Trigo, CIMMYT) as well as South Asia were found to be superior general combiners for grain filling duration. Likewise, crosses involving diverse parents from CIMMYT and South Asia showed significant SCA effects for grain filling duration and other traits. |
Plant Genetics Combining ability of elite clones of Eucalyptus grandis and Eucalyptus urophylla with Eucalyptus globulus Bison, Odair Ramalho, Magno Antonio Patto Rezende, Gabriel Dehon Sampaio Peçanha Aguiar, Aurélio Mendes Resende, Marcos Deon Vilela de Abstract in English: In Brazil, eucalyptus breeding programs for cellulose production has used two species, Eucalyptus grandis and Eucalyptus urophylla. Nevertheless, it would be useful to introgress alleles from other species to improve wood quality and volume. The objective of this research was to evaluate the hybrid potential of elite clones of E. grandis and E. urophylla from the Aracruz Celulose Company S. A. with Eucalyptus globulus clones. To do so, six elite clones were crossed with ten E. globulus clones in a half-diallel mating design. The resulting hybrid combinations as well as the four check clones were evaluated in randomized complete block experiments with single plant plots and 40 replicates from September to October 2001 at three Brazilian sites, Aracruz and São Mateus in the Espírito Santo state and Caravelas in Bahia State. Two years later the circumference at breast height (CBH) and the wood density (WD) were measured. The means were submitted to diallel analysis according to the Griffing method (1956), adapted by Geraldi and Miranda Filho (1988). Although the number of clones involved was small, the crossings of elite clones of E. grandis and E. urophylla with clones of E. globulus were promising, especially for wood quality gains. |
Plant Genetics PCR-RFLP analysis of non-coding regions of cpDNA in Araucaria angustifolia (Bert.) O. Kuntze Schlögl, Paulo Sérgio Souza, Anete Pereira de Nodari, Rubens Onofre Abstract in English: The Araucaria angustifolia (Bert.) O. Kuntze, also named the "paraná pine" (‘pinheiro-do-Paraná’ in Portuguese), is a native conifer species naturally occurring in the Brazilian Tropical Atlantic Forest which in Brazil is mostly limited to the southern Brazilian states of Paraná, Santa Catarina and Rio Grande do Sul. Chloroplast DNA markers (cpDNA) are useful in populational genetic studies because of their low substitution rate and the uniparental transmission. The conservation of cpDNA genes between species has allowed the design of consensus chloroplast primers that have had a great impact on population genetics and phylogenetic studies. In this study we used the polymerase chain reaction technique combined with restriction enzyme fragment length polymorphism (PCR-RFLP) to characterize the genetic diversity of the chloroplast genome in nine natural A. angustifolia populations. Among the 141 trees surveyed we found 12 different cpDNA haplotypes and demonstrated that A. angustifolia has high levels of total diversity (hT = 0.612) and an average within-population diversity (hS) of 0.441, suggesting the presence of high within-population variation. The estimated genetic divergence could be helpful in designing breeding programs and species conservation strategies, although additional studies with a larger number of populations and trees is essential for a better understanding of gene flow and the inheritance of major Araucaria angustifolia traits. |
Plant Genetics Diallel analysis of fruit set in Hevea brasiliensis Muell. Arg. Omokhafe, Kenneth O. Akpobome, Frederick A. Nasiru, Ibrahim Abstract in English: The clonal and inter-clonal combining ability of fruit set in a Hevea brasiliensis four-parent diallel mating was evaluated using a randomized complete block experimental design with three replicates. Twelve main and reciprocal crosses were hand pollinated and percentage fruit set was recorded. The raw data were subjected to an arc-sine transformation for analysis of variance, and general and specific combining ability. There was significant variation of each of fruit set for the various crosses, general combining ability and reciprocal effect. The breeding implications of these results are also discussed. |
Genetics of Microorganisms Quantitative real-time PCR for the clinical detection of Helicobacter pylori Ribeiro, Marcelo Lima Ecclissato, Christina Cunha Mattos, Ricardo Gabriel Mendonca, Sergio Pedrazzoli Jr., José Abstract in English: Accurate diagnosis of Helicobacter pylori infection is very important in both clinical practice and research. We evaluated the sensitivity of real-time PCR (RT-PCR) for the detection and quantification of Helicobacter pylori using DNA from 91 human gastric biopsy samples divided into three groups: 46 biopsies from untreated patients who according to the references methods were considered H. pylori-negative (group A); 35 biopsies from patients previously treated against H. pylori and considered to be cured by "gold standard" tests (group B); and 10 biopsies from patients H. pylori-positive by all available methods (group C). The sensitivity of the RT-PCR assay was higher than that of standard methods. Of the 81 patients considered to be uninfected according to the references methods, 16 were H. pylori-positive by PCR, 10 of which were patients who had received H. pylori eradication therapy and 6 were untreated patients. Based on these findings we recommend that RT-PCR should be use in addition to standard methods in clinical studies to monitor the results of H. pylori eradication therapy. |
Mutagenesis Cytotoxic and genotoxic potential of surface water from the Pitimbu river, northeastern/RN Brazil Egito, Lucila Carmem Monte Medeiros, Maria das Graças Medeiros, Silvia Regina Batistuzzo de Agnez-Lima, Lucymara Fassarella Abstract in English: In this study, the onion (Allium cepa) root test was used to evaluate the genotoxicity of the Pitimbu River (Natal city, Brazil) surface water. The water was collected at five sampling sites along the river and one sample was obtained after the treatment (flocculation, chlorination and pH correction) of the river water for human consumption. All raw river water samples increased the frequency of chromosomal abnormalities and/or micronuclei and two of the water samples produced alterations in the mitotic index of the root cells. Two of the water samples also altered root growth and two produced morphological modifications in the A. cepa roots. Water collected from a site near an industrial area was the most consistently toxic and genotoxic of the samples. Although the water chlorinated for human consumption was not genotoxic, the data indicate that surface water from the Pitimbu River contains toxic and genotoxic compounds that potentially may impact this aquatic ecosystem. |
Mutagenesis Anticlastogenic potential and antioxidant effects of an aqueous extract of pulp from the pequi tree (Caryocar brasiliense Camb) Khouri, Juliana Resck, Inês S. Poças-Fonseca, Marcio Sousa, Thiago M.M. Pereira, Luciana O. Oliveira, Adriano B.B. Grisolia, Cesar K. Abstract in English: The effectiveness of an aqueous extract of Caryocar brasiliense (Caryocaraceae) Camb pulp, popularly known in Brazil as pequi, against clastogenicity induced by cyclophosphamide and bleomycin was evaluated using an in vivo mouse bone marrow cell micronuclei test, an in vitro Chinese hamster ovary cell (CHO-K1) chromosome aberration test and an in vitro antioxidant assay based on the oxidative damage to 2-deoxy-D-ribose (2-DR) induced by hydroxyl radicals (•OH) generated by the reaction between ascorbic acid and (Fe III)-EDTA. In mouse bone marrow cells the extract showed a protective effect against micronuclei induced by cyclophosphamide and bleomycin but did not interfere with polychromatic bone marrow erythrocyte proliferation, except when the mice had been treated with the highest dose of cyclophosphamide. When CHO-K1 cells were pretreated by adding 0.01, 0.05 or 0.1 mL of extract per mL of cell culture medium 24 or 48 h before bleomycin or cyclophosphamide there was a protective effect against chromosome breaks and a significant decrease in the mitotic index (a measure of cytotoxicity) of the CHO-K1 cells. The extract also had a protective effect against oxidative hydroxyl radical damage to 2-DR. This study suggests that C. brasiliense pulp aqueous extract has anticlastogenic potential, possibly due to its antioxidative properties. |
Mutagenesis Modulatory effects of the antioxidant ascorbic acid on the direct genotoxicity of doxorubicin in somatic cells of Drosophila melanogaster Fragiorge, Edson José Spanó, Mário Antônio Antunes, Lusânia Maria Greggi Abstract in English: In this study two different crosses involving the wing cell markers mwh and flr³ (standard (ST) cross and high bioactivation (HB) cross, the latter being characterized by a high constitutive level of cytochrome P450 which leads to an increased sensitivity to a number of promutagens and procarcinogens) were used to investigate the modulatory effects of ascorbic acid (AA) combined with the antitumor agent doxorubicin (DXR) in Drosophila melanogaster. We observed that the two different concentrations of AA (50 or 100 mM) had no effect on spots frequencies, while DXR treatments (0.2 or 0.4 mM) gave positive results for all types of spots, when compared to negative control. For marker-heterozygous (MH) flies, a protective effect was observed with the lower concentration of AA (50 mM) that was able to statistically decrease the frequency of spots induced by DXR (0.2 mM), while an enhanced frequency of spots induced by DXR was observed with the higher concentration of AA (100 mM), when compared to DXR treatment (p < 0.05). These results suggest that AA may interfere with free radicals generated by DXR and with other possible reactive metabolites. The efficiency of AA in protecting the somatic cells of D. melanogaster against mutation and recombination induced by DXR is dependent on the dose used and the protection is directly related to the activity of cytochrome P450 enzymes. |
Mutagenesis Evaluation of DNA damage in an Ecuadorian population exposed to glyphosate Paz-y-Miño, César Sánchez, María Eugenia Arévalo, Melissa Muñoz, María José Witte, Tania De-la-Carrera, Gabriela Oleas Leone, Paola E. Abstract in English: We analyzed the consequences of aerial spraying with glyphosate added to a surfactant solution in the northern part of Ecuador. A total of 24 exposed and 21 unexposed control individuals were investigated using the comet assay. The results showed a higher degree of DNA damage in the exposed group (comet length = 35.5 µm) compared to the control group (comet length = 25.94 µm). These results suggest that in the formulation used during aerial spraying glyphosate had a genotoxic effect on the exposed individuals. |
Evolutionary Genetics Research Article Comparing covariance matrices: random skewers method compared to the common principal components model Cheverud, James M. Marroig, Gabriel Abstract in English: Comparisons of covariance patterns are becoming more common as interest in the evolution of relationships between traits and in the evolutionary phenotypic diversification of clades have grown. We present parallel analyses of covariance matrix similarity for cranial traits in 14 New World Monkey genera using the Random Skewers (RS), T-statistics, and Common Principal Components (CPC) approaches. We find that the CPC approach is very powerful in that with adequate sample sizes, it can be used to detect significant differences in matrix structure, even between matrices that are virtually identical in their evolutionary properties, as indicated by the RS results. We suggest that in many instances the assumption that population covariance matrices are identical be rejected out of hand. The more interesting and relevant question is, How similar are two covariance matrices with respect to their predicted evolutionary responses? This issue is addressed by the random skewers method described here. |
Evolutionary Genetics Allozyme variation and sociogenetic structure of Polistes satan Bequaert 1940 colonies (Hymenoptera, Vespidae) Gaspar, Jane Cristina Wehren López-Uribe, Margarita María Del Lama, Marco Antônio Abstract in English: We collected 40 colonies of the wasp Polistes satan in November 1993 (Population A = 23 colonies) and October 1995 (Population B = 17 colonies) from the town of Delfinopólis in the Brazilian state of Minas Gerais. Pupae from these nests were used to assess allozyme variation and genetic relatedness among nestmates. Of the 24 gene loci sampled six showed allozyme polymorphism (Est-2, Pgm-1, 6Pgd, Hk-1, Idh and Pep-A) and all loci except Idh were at Hardy-Weinberg equilibrium. Populations A and B showed distinct intra-loci heterozygosities, but similar average heterozygosity over all loci. Significant inter-population differentiation (Fst = 0.051; p < 0.01) was detected, probably due to genetic drift driven by bottlenecking that occurred due to the first sampling. Mean intra-colony relatedness values between female broods (rA = 0,525 ± 0.063; rB = 0,456 ± 0.103) indicated that P. satan has a simple sociogenetic structure where one female takes on most of the reproductive tasks but, eventually, subordinates also reproduce in the colony. |
Evolutionary Genetics Molecular evidence of polyphyletism in the plant genus Carum L. (Apiaceae) Papini, Alessio Banci, Francesca Nardi, Enio Abstract in English: An analysis of internal transcribed spacer (ITS) DNA sequences of the four species of Carum L. (Apiaceae) known in Italy revealed that this genus is polyphyletic. Maximum parsimony with bootstrap resampling, maximum likelihood and Bayesian inference analyses resulted in three distinct clades: Carum carvi L. clustered within tribe Careae Baill. (former Aegopodium clade); Hellenocarum multiflorum (= Carum multiflorum), Carum heldreichii and Carum appuanum clustered within the tribe Pyramidoptereae Boiss.; and H. multiflorum and C. heldreichii formed a well supported clade. Since the sister group of H. multiflorum and C. heldreichii was Bunium elegans the autonomy of Hellenocarum from Carum is confirmed by our study. We also found that C. appuanum clustered separately from the other Carum species, with the closest related species appearing to be Scaligeria moreana but this still had few morphological similarities with C. appuanum. |
Developmental Genetics Connexins in the early development of the African clawed frog Xenopus laevis (Amphibia): The role of the connexin43 carboxyl terminal tail in the establishment of the dorso-ventral axis Cofre, Jaime Abdelhay, Eliana Abstract in English: Connexins are a family of related proteins identified in vertebrate forming gap junctions, which mediate cell-to-cell communication in early embryos, with an important role in establishing embryonic asymmetry and ‘communication compartments’. By in situ hybridization, immunocytochemistry, reverse transcriptase PCR (RT-PCR) and western blotting we show that a Cx43-like molecule is present in oocytes and embryos of the African clawed frog Xenopus laevis, with specific localization in the animal-vegetal axis. This specific distribution is suggestive for an important role for this protein in the establishment of the dorso-ventral axis. Antisense RNA and antibodies directed against rat carboxyl terminal tail of the Cx43 (CT-Cx43) and injected in 1-cell stage Xenopus embryos, induced pronounced alterations in nervous system development, with a severe ventralization phenotype. Coherently, the overexpression of CT-Cx43 produced a dorsalization of the embryos. In antisense treated embryos, the expression of the beta-catenin gene is eliminated from the Nieuwkoop center, the pattern expression of the Chordin, Xnot and Xbra is modified, with no effect in expression of the Goosecoid gene. In CT-Cx43 mRNA treated embryos the pattern of expression of the beta-catenin, Chordin, Goosecoid, Xnot and engrailed-2 genes is modified. The expression of beta-catenin is increased in the Nieuwkoop center, the expression pattern of Chordin and Goosecoid is expanded to the posterior neural plate and engrailed-2 presents ectopic expression in the ventral region. Taken together our data suggest a role for CT-Cx43 as a maternal determinant with a critical function in the formation of the dorso-ventral axis in Xenopus laevis. The Cx43 may be one of the earliest markers of the dorso-ventral axis in these embryos and could possibly be acting through regionalization of factors responsible for the establishment of this axis. |
Developmental Genetics Characterization of partial Hox gene sequences in annual fish of the subfamily Cynolebiatinae (Cyprinodontiformes, Rivulidae) Gutiérrez, Verónica Arezo, María José García, Graciela Abstract in English: Hox genes encode a family of transcription factors implicated in conferring regional identity along the anteroposterior axis in developing animal embryos. These genes are organized in genomic clusters, expressed collinearly and highly conserved in vertebrates. Among teleost, South American annual killifishes of the Cynolebiatinae subfamily represent an excellent model in development studies because their embryos are capable of undergoing reversible developmental arrest (diapause) at three well-defined morphological stages. They are also an excellent model for evolutionary studies due to the high rates of mutation of their mitochondrial genome, their karyotypic divergence and their morphological variability. In this study, three partial homeobox sequences were isolated from different species of the Cynolebiatinae subfamily. Phylogenetic analyses and sequence comparisons revealed that they belong to the anterior Hox complex group, specifically to paralogue groups 1 and 3. This is the first time that partial Hox genes have been described in species of the Cynolebiatinae subfamily. |