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Genetics and Molecular Biology, Volume: 31, Número: 4, Publicado: 2008
  • Linkage disequilibrium, genetic association mapping and gene localization in crop plants

    Sorkheh, Karim; Malysheva-Otto, Lyudmyla V.; Wirthensohn, Michelle G.; Tarkesh-Esfahani, Saeed; Martínez-Gómez, Pedro

    Resumo em Inglês:

    DNA-based molecular markers have been extensively utilized for a variety of studies in both plant and animal systems. One of the major uses of these markers is the construction of genome-wide molecular maps and the genetic analysis of simple and complex traits. However, these studies are generally based on linkage analysis in mapping populations, thus placing serious limitations in using molecular markers for genetic analysis in a variety of plant populations. Therefore, alternative approach has been suggested, linkage disequilibrium-based association analysis which detects and locates quantitative trait loci (QTL) by the strength of the correlation between a trait and a marker. Although association analysis has already been used for studies on genetics of complex traits in humans, its use in plants has newly started. In the present review, we describe what is known about variation in linkage disequilibrium (LD) and summarize published results on association studies in crop plant species. We give a list of different factors affecting LD, and discuss the current issues of LD research in plants. Later, we also describe the various uses of LD in crop plants research and summarize the present status of LD researches in different plant genomes. Finally, future key issues about the application of these studies on the localization of genes in these crop plants have been also discussed.
  • HOX gene analysis in the osteogenic differentiation of human mesenchymal stem cells Human And Medical Genetics

    Chae, Song Wha; Jee, Bo Keun; Lee, Joo Yong; Han, Chang Whan; Jeon, Yang-Whan; Lim, Young; Lee, Kweon-Haeng; Rha, Hyoung Kyun; Chae, Gue-Tae

    Resumo em Inglês:

    Human bone marrow-derived mesenchymal stem cells (hMSCs) have the capacity to differentiate into osteoblasts during osteogenesis. Several studies attempted to identify osteogenesis-related genes in hMSCs. Although HOX genes are known to play a pivotal role in skeletogenesis, their function in the osteogenesis of hMSCs has not yet been investigated in detail. Our aim was to characterize the expression of 37 HOX genes by multiplex RT-PCR to identify the ones most probably involved in osteogenic differentiation. The results showed that the expression patterns of four HOX genes were altered during this process. In particular, the expression levels of HOXC13 and HOXD13 were dramatically changed. Real-time PCR and Western blot analysis were performed in order to further analyze the expression of HOXC13 and HOXD13 . The qRT-PCR results showed that transcription of HOXC13 was up-regulated by up to forty times, whereas that of HOXD13 was down-regulated by approximately five times after osteogenic differentiation. The Western blot results for the HOXC13 and HOXD13 proteins also corresponded well with the real-time PCR result. These findings suggest that HOXC13 and HOXD13 might be involved in the osteogenic differentiation of hMSCs.
  • Autonomous growth of BALB/MK keratinocytes transfected with a retroviral vector carrying the human epidermal growth factor gene Human And Medical Genetics

    Choudhuri, Jomuna V.; Mathor, Monica B.; Silva, Flávia H.; Han, Sang W.

    Resumo em Inglês:

    Epidermal growth factor (EGF), which promotes epidermal regeneration and wound closure, is important for the proliferation and differentiation of epidermal and epithelial tissues in animals. Exogenous EGF is a promising therapeutic agent for wound healing, but its general use is restricted by the limited availability of this protein. In this work, we show that the transfection of mouse BALB/MK keratinocytes, which are totally dependent on EGF for growth and migration, with mature cDNA for human EGF via a retroviral vector abolished the cells requirement for exogenous EGF. The transformed cells had normal morphology and a growth rate that varied according to the source of the retroviral vector used. Keratinocyte transfection with EGF cDNA provides a time- and cost-efficient means of culturing keratinocytes and yields cells that may be useful for skin grafting.
  • Prenatal diagnosis of fetal chromosomal abnormalities: report of an 18-year experience in a Brazilian public hospital Human And Medical Genetics

    Kessler, Rejane G.; Sanseverino, Maria Teresa V.; Leistner-Segal, Sandra; Magalhães, José A.A.; Giugliani, Roberto

    Resumo em Inglês:

    The study of the fetal karyotype became an important tool for the fetal diagnosis of genetic diseases in the 1970s. Although application of this test has remained very restricted in Brazil, we had 905 referrals for prenatal fetal karyotyping between 1989 and 2007. In 879 cases, a fetal karyotype was obtained. We detected 74 abnormal karyotypes (8.4%), the majority being found when the prior indication was fetal malformation. When obtaining amniotic fluid or chorionic villus samples was difficult, alternative fetal materials (urine, cystic hygroma, cystic lung, intreperitoneal and cerebrospinal fluids) were collected and we had success in obtaining karyotypes in all 13 cases. Although, the option of terminating abnormal pregnancies does not legally exist in Brazil, the information gained in assessing the prognosis of on-going pregnancies or estimating recurrence risks justifies prenatal diagnosis of chromosome abnormalities. We conclude that, in keeping with the policy in most other countries, prenatal cytogenetic analysis is strongly recommended in high-risk pregnancies for fetal abnormalities. However, the unique aspect of this type of study is not its rarity in world terms, but its rarity in Brazil. This argues that Brazilian health policy on prenatal diagnosis requires reforming to make it much more widely available within the public health care sector.
  • Editorial commentary on the state of clinical genetics in Brazil exemplified by the article on prenatal diagnosis by Kessler et al. Genetics and Molecular Biology (this issue) Human And Medical Genetics

    Pearson, Peter
  • MTHFR , prothrombin and Factor V gene variants in Turkish patients with coronary artery stenosis Human And Medical Genetics

    Caner, Müge; Bircan, Rifat; Sevinç, Deniz; Benli, Fehime; Güney, A. Ilter; Kurtoglu, Nuri

    Resumo em Inglês:

    Many epidemiological studies have reported an association between hemostatic factors and risk of both coronary and peripheral artery diseases. Using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis, we investigated the association between coronary artery disease and polymorphisms in the methylenetetrahydrofolate reductase ( MTHFR C677T and A1298C), prothrombin (G20210A), and factor V (A4070G) genes. We screened these gene variants in 174 subjects who had undergone coronary angiography - 115 patients with patent coronary artery disease (grade 3 vessel disease, i.e. , significant coronary stenosis), and 59 healthy controls with grade 0 vessel disease. The analysis of our data did not show any statistically significant association between coronary artery disease (CAD) and the investigated polymorphisms.
  • An illustrative case of Léri-Weill dyschondrosteosis Human And Medical Genetics

    Lima, Renata de; Iamada, Cristina Forti; Silva, Luciana Oliveira; Mello, Maricilda Palandi de; Maciel-Guerra, Andréa Trevas

    Resumo em Inglês:

    We report on a girl presenting Léri-Weill dyschondrosteosis (LWD) due to deletion of the SHOX gene. Her family included individuals with short stature alone or with both short stature and mesomelia or Madelung's deformity. The deletion was demonstrated through detection of hemizygosity for microsatellite markers SHOX-CA repeat, DXYS10092, DXYS10093 and DXYS10091 localized around the SHOX gene, with retention of paternal alleles in the proband and three of her sisters who had short stature as the only clinical feature. Hemizygosity for these loci was also observed in their mother, who had short stature too. The deletion in the proband was however larger, including locus DXY10083. The proband's only sister with normal height did not carry the deletion. Family history suggests transmission of the deletion from the proband's maternal great-grandfather to her grandfather via the Y chromosome, and from the grandfather to the proband's mother via the X chromosome after crossing-over in the pseudoautosomal region proximal to the SHOX gene.
  • Association of a locus on rat chromosome 4 with anxiety-related behaviors in two selectively bred rat lines Animal Genetics

    Hameister, Thaïs M.; Izídio, Geison S.; Valiati, Victor H.; Ramos, André

    Resumo em Inglês:

    The Floripa H and L rat lines, selected for high and low locomotion in the central aversive area of an open field, a widely used emotionality test, were proposed as a model for studying the genetic basis of anxiety. The present study aimed to verify if the QTL Ofil1, mapped to rat chromosome 4 and previously identified as being related to emotionality in another population of rats, contributes to the behavioral variability observed in the Floripa rat lines. To this purpose, rats of five generations of selective breeding were genotyped for two polymorphic markers, D4RAT59 and D4MGH27, flanking Ofil1. Changes in genotype and allele frequencies throughout generations were evaluated in both H and L lines, in order to assess if the bidirectional selection based on behavioral scores induced divergent changes in the genotype of this genome region. There were significant changes in genotype frequencies for both molecular markers, however, only the genotype variations of the D4RAT59 marker were significantly correlated with the variations in the selected phenotype. This result suggests that the region of the genome near D4RAT59 contains one or more genes contributing to the interindividual variation in central locomotion in the open field test.
  • Artificial family selection based on growth rate in cultivated lines of Litopenaeus vannamei (Decapoda, Penaeidae) from Venezuela Animal Genetics

    De Donato, Marcos; Ramirez, Raúl; Howell, Chris; Verginelli, Roberto; Orta, Tomás; Cabrera, Saúl; Mata, Enrique; Manrique, Ramón

    Resumo em Inglês:

    We investigated the effects of family selection on the growth rates of four genetic lines of Litopenaeus vannamei, the Pacific white shrimp, from Venezuela. The mean family weight at selection was 16.79 g and the mean growth rate was 0.169 g d-1 at a mean age of 138 days. The mean growth rates per generation were 0.141 g d-1 for the parental generation (P0), 0.173 g d-1 for the F1 and 0.191 g d-1 for the F2. Survival varied from 83.3% to 94.4%. There were statistically significant differences between lines, with lines B and C, growing at higher temperatures and lower salinities, showing the fastest growth. Regression analysis between growth rate and age for each genetic line showed high r² values, but smaller than those shown between growth rate and generation. The mean heritability (h²) estimated for growth rate was 0.25 ± 0.04, with line per generation values ranging from 0.18 to 0.38. The mean heritability for growth rate per line showed some variation but there was no correlation between heritability and growth rate. Females showed a greater gain in growth rate per generation than males due to their faster growth when they were older. This study shows the high potential of these L. vannamei populations for the future of shrimp culture in the Americas.
  • Cytogenetic studies of three Lycosidae species from Argentina (Arachnida, Araneae) Animal Genetics

    Chemisquy, María A.; Gil, Sergio G. Rodríguez; Scioscia, Cristina L.; Mola, Liliana M.

    Resumo em Inglês:

    Cytogenetic studies of the family Lycosidae (Arachnida: Araneae) are scarce. Less than 4% of the described species have been analyzed and the male haploid chromosome numbers ranged from 8+X 1 X 2 to 13+X 1 X 2 . Species formerly classified as Lycosa were the most studied ones. Our aim in this work was to perform a comparative analysis of the meiosis in " Lycosa " erythrognatha Lucas, " Lycosa " pampeana Holmberg and Schizocosa malitiosa (Tullgren). We also compared male and female karyotypes and characterized the heterochromatin of " L. " erythrognatha . The males of the three species had 2n = 22, n = 10+X 1 X 2 , all the chromosomes were telocentric and there was generally a single chiasma per bivalent. In " Lycosa " pampeana , which is described cytogenetically for the first time herein, the bivalents and sex chromosomes showed a clustered arrangement at prometaphase I. The comparison of the male/female karyotypes (2n = 22/24) of " Lycosa " erythrognatha revealed that the sex chromosomes were the largest of the complement and that the autosomes decreased gradually in size. The analysis of the amount, composition and distribution of heterochromatin with C-banding and staining with DAPI- and CMA 3 - showed that " Lycosa " erythrognatha had little GC-rich heterochromatin in the pericentromeric region of all chromosomes. In addition, the actual occurrence of the genus Lycosa in the Southern Hemisphere is discussed.
  • Mapping of ribosomal genes and chromosomal markers in three species of the genus Serrasalmus (Characidae, Serrasalminae) from the Amazon basin Animal Genetics

    Nakayama, Celeste M.; Feldberg, Eliana; Bertollo, Luiz Antonio C.

    Resumo em Inglês:

    Karyotypic characteristics of three species of the genus serrasalmus ( S. altispinnis , S. gouldingi and S. Serrulatus ) from the middle and lower Negro River, Amazon Basin, were investigated using different staining techniques and Fluorescent in situ hybridization with 5S and 18S rDNA probes. The diploid number was invariably 2n = 60 and the fundamental number was FN = 110. Nevertheless, the karyotypes differed from each other in composition: 24m, 20sm, 6st, 10a in S. altispinnis ; 22m, 22sm, 6st, 10a in S. gouldingi and 20m, 22sm, 8st, 10a in S. serrulatus. The karyotype of S. altispinnis differed from the one previously described in a population from the Pitinga River. C-positive constitutive heterochromatin was mainly pericentromeric in the karyotypes of all species. Nucleolar organizer regions were multiple and preferentially located terminally on the short arms of the subtelocentric/acrocentric chromosomes, as evidenced by both silver nitrate staining and fluorescent in situ hybridization with the 18S rDNA probe. The maximum number of NORs varied among species, as did the NOR-bearing chromosomes. FISH with the 5S rDNA probe produced an interstitial signal on the long arms of the pair 7 in all species, coincident with a C-positive heterochromatic band. While some chromosome features were shared by the three species, some were species-specific and thus useful for cytotaxonomy.
  • Growth hormone 1 gene (GH1) polymorphisms as possible markers of the production potential of beef cattle using the Brazilian Canchim breed as a model Animal Genetics

    Silveira, Luiz Guilherme G.; Furlan, Luiz R.; Curi, Rogério A.; Ferraz, André Luiz J.; Alencar, Maurício M. de; Regitano, Luciana C.A.; Martins, Cyntia L.; Arrigoni, Mário de Beni; Suguisawa, Liliane; Silveira, Antônio C.; Oliveira, Henrique N. de

    Resumo em Inglês:

    The growth hormone 1 gene (GH1) is a candidate gene for body weight and weight gain in cattle since it plays a fundamental role in growth regulation. We investigated the GH1 gene AluI and DdeI restriction enzyme polymorphisms, located 149 bp apart in the cattle genome, as possible markers of the production potential of Canchim crossbreed cattle, a 5/8 Charolais (Bos taurus) and 3/8 Nelore (Bos indicus) breed developed in Brazil, by evaluating the birth weight, weaning weight, yearling weight and plasma insulin-like growth factor-1 (IGF-1) concentration of 7 month to 10 months old Canchim calves (n = 204) of known genealogy and which had been genotyped for the AluI and DdeI markers. Our results showed significant effect (p < 0.05) between the homozygous DdeI+/DdeI+ polymorphism and the estimated breeding value for weaning weight (ESB-WW), while the AluI leucine homozygous (L/L) and leucine/valine (L/V) heterozygous polymorphisms showed no significant effect on the traits studied. The restriction sites of the two enzymes led to the formation of haplotypes which also exerted a significant effect (p < 0.05) on the ESB-WW, with the largest difference being 8.5 kg in favor of the homozygous L plus DdeI+/L plus DdeI+ genotype over the heterozygous L plus DdeI-/V plus DdeI+ genotype.
  • Polymorphism of locus DRB3.2 in populations of Creole Cattle from Northern Mexico Animal Genetics

    Fernández, Ilda G.; Ramírez, José Gonzalo Ríos; Vázquez, Amanda Gayosso; Arvizu, Raúl Ulloa; Morales, Rogelio A. Alonso

    Resumo em Inglês:

    The polymorphism of locus BoLA-DRB3.2 of the Major Histocompatibility Complex was evaluated in two northern Mexican Creole cattle populations, Chihuahua (n = 47) and Tamaulipas (n = 51). The BoLA-DRB3.2 locus was typed by amplification and digestion with restriction endonuclease enzymes (PCR-RFLP). Fifty-two alleles were detected (28 previously reported and 24 new ones). In the Chihuahua population, 18 alleles and 5.5 effective alleles were found, while in the Tamaulipas population there were 34 and 10.8, respectively. The allele frequencies ranged from 0.011 to 0.383 in Chihuahua and from 0.010 to 0.206 in Tamaulipas. The frequencies of the new alleles in both cattle populations were low (0.010 to 0.053). The expected heterozygosity was 0.827 and 0.916, respectively, for the Chihuahua and Tamaulipas populations. Both populations presented a heterozygote deficit: [Chihuahua F IS = 0.1 (p = 0.019) and Tamaulipas F IS = 0.317 (p < 0.001)]. In conclusion, this study showed that the Mexican Creole cattle have many low-frequency alleles, several of which are exclusive to these populations. Genetic distances obtained show that the Mexican Creole cattle population is composed of independent populations, far apart from other South American Creole populations.
  • Karyotype characterization and nucleolar organizer regions of marsupial species (Didelphidae) from areas of Cerrado and Atlantic Forest in Brazil Animal Genetics

    Pereira, Núbia P.; Ventura, Karen; Silva Júnior, Magno C.; Silva, Daniela de Melo e; Yonenaga-Yassuda, Yatiyo; Pellegrino, Katia C. M.

    Resumo em Inglês:

    The karyotypes of 23 specimens belonging to 16 species from nine genera of Brazilian marsupials (family Didelphidae) were studied. The animals were collected in eight localities of Cerrado or Atlantic Forest biomes in the states of Goiás, Tocantins and São Paulo. The karyotypes were analyzed after conventional Giemsa staining and silver staining of the nucleolus organizer regions (Ag-NORs). New karyotypic data were obtained for Gracilinanus microtarsus (2n = 14, FN = 24), Marmosops paulensis (2n = 14, FN = 24) , Micoreus paraguayanus (2n = 14, FN = 20) and Monodelphis rubida (2n = 18, FN = 32) and are discussed in detail. The karyotypes of G. microtarsus , M. paulensis and M. paraguayanus include three large pairs of submetacentrics (pairs 1, 2 and 3) and a medium-sized metacentric or submetacentric pair 4. Pairs 5 and 6 are small submetacentrics in G. microtarsus and M. paulensis and acrocentrics in M. paraguayanus . M. paulensis presented a single Ag-NOR in pair 6 (6p6p), while M. paraguayanus exhibited multiple Ag-NORs in pairs 5 and 6 (5pq5pq6p6p). There was variation in size and morphology of the sex chromosomes among these species. Monodelphis rubida presented a karyotype with 2n = 18 and FN = 32 composed of a large submetacentric pair 1, a medium-sized metacentric pair 2 and six pairs of submetacentrics (pairs 3 through 8). The X was a small acrocentric and the Y was dot-like. A single Ag-NOR bearing pair (5p5p) characterized M. rubida. Relevant karyotypic information was obtained for 19 specimens belonging to 12 species collected in areas sampled for the first time [ Caluromys lanatus and C. philander (2n = 14, FN = 20), Gracilinanus emiliae (2n = 14, FN = 24), Marmosa murina , Metachirus nudicaudatus and Micoureus demerarae (2n = 14, FN = 20), Monodelphis americana (2n = 18, FN = 32) and M. domestica (2n = 18, FN = 20), and Didelphis marsupialis, Philander frenata, P. opossum and P. sp (2n = 22, FN = 20)]. Although the karyotypes were relatively conserved with respect to the morphology of the autosomes among species with the same diploid number, some differences regarding FN, sex chromosomes morphology and Ag-NORs patterns were detected.
  • Bovine kappa-casein gene polymorphism and its association with milk production traits Animal Genetics

    Rachagani, Satyanarayana; Gupta, Ishwar Dayal

    Resumo em Inglês:

    Point mutations in exon IV of the bovine κ-casein (CSN3) gene determine two allelic variants, A and B. These variants were distinguished by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis in the indigenous Sahiwal and Tharparkar cattle breeds. DNA samples (252 Sahiwal and 56 Tharparkar) were analyzed for allelic variants of the CSN3 gene. Polymorphism was detected by digestion of PCR-amplified products with HindIII, HhaI and HaeIII restriction enzymes, followed by separation on 3% agarose gels, and resolved by ethidium bromide staining. Allele A of the κ-casein gene occurred at a higher frequency than allele B, in both Sahiwal and Tharparkar breeds. The genotypic frequencies of AA, AB, and BB in the Sahiwal and Tharparkar breeds were 0.758, 0.230 and 0.012, and 0.0.732, 0.250 and 0.018, respectively. The frequencies of alleles A and B in the Sahiwal and Tharparkar breeds were 0.873 and 0.127, and 0.857 and 0.143, respectively. Genotype BB of the kappa-casein gene had more influence on the monthly milk yield, 305-days milk yield, monthly solids-not-fat (SNF) yield, and monthly protein yield, in the Sahiwal cattle.
  • Quantitative trait loci for carcass, internal organ and meat quality traits on porcine chromosomes 16, 17 and 18 Animal Genetics

    Paixão, Débora M.; Silva Filho, Miguel I. da; Pereira, Mário S.; Lopes, Marcos S.; Barbosa, Leandro; Souza, Katiene Régia Silva; Lopes, Paulo S.; Guimarães, Simone E.F.

    Resumo em Inglês:

    The objective of this study was to map quantitative trait loci (QTL) on porcine chromosomes 16, 17 and 18 and to determine their association with carcass, organ and meat quality traits. An F2 population was produced by crossing two boars of the naturalized Brazilian Piau breed with 18 commercial females (Landrace x Large White x Pietrain). The population was genotyped for 11 microsatellite markers distributed over the three chromosomes and the results were used to construct a marker-specific linkage map for the population. Analysis of the polymorphic information content showed that the microsatellite markers were adequate for the study of quantitative traits. QTL were identified by regression interval mapping using QTL Express software. QTL not previously described in the literature were detected on chromosome 16, whereas QTL described in other populations were detected on chromosomes 17 and 18. The information from the significant QTL identified here will be useful for future fine-mapping studies and should provide a better understanding of productive phenotypes in pigs.
  • β-casein gene polymorphism permits identification of bovine milk mixed with bubaline milk in mozzarella cheese Animal Genetics

    Otaviano, Antonio R.; Lima, André Luis F.; Laureano, Monyka M.M.; Sena, Janete A.D.; Albuquerque, Lúcia G. de; Tonhati, Humberto

    Resumo em Inglês:

    Mozzarella cheese is traditionally prepared from bubaline (Bubalus bubalis) milk, but product adulteration occurs mainly by addition of or full substitution by bovine milk. The aim of this study was to show the usefulnes of molecular markers to identify the admixture of bovine milk to bubaline milk during the manufacturing process of mozzarella cheese. Samples of mozzarella cheese were produced by adding seven different concentrations of bovine milk: 0%, 1%, 2%, 5%, 8%, 12% and 100%. DNA extracted from somatic cells found in cheese were submitted to PCR-RFLP analysis of casein genes: α-s1-CN - CSN1S1 that encompasses 954 bp from exon VII to intron IX (Alu I and Hinf I), β-CN - CSN2 including 495 bp of exon VII (Hae III and Hinf I), and κ-CN - CSN3, encompassing 373 bp of exon IV (Alu I and Hind III). Our results indicate that Hae III-RFLP of CSN2 exon VII can be used as a molecular marker to detect the presence of bovine milk in "mozzarella" cheese.
  • Nonparametric stability methods for interpreting genotype by environment interaction of bread wheat genotypes (Triticum aestivum L.) Plant Genetics

    Akcura, Mevlut; Kaya, Yuksel

    Resumo em Inglês:

    Evaluation of performance stability and high yield is essential for yield trials conducted in different environments. We determined the stability of 10 bread wheat (Triticum aestivum L.) genotypes (5 cultivars and 5 advanced lines) using nonparametric stability measures and compared nonparametric stability statistics across 19 environments in the Central Anatolian Region of Turkey. Experiments were setup in a randomized complete block design with four replications. Nonparametric stability measures revealed that AL-2, Karahan-99 and Altay-2000 were the most stable and well adapted genotypes across the 19 environments. In addition, it was concluded that graphs of mean grain yield (t ha-1) vs. nonparametric measures (Si(¹), Si(²), S(³), Si(6) and σgy) values and kr vs. σr values could enhance visual efficiency of genotype selection based on genotype by environment interaction. Furthermore, high TOP values, Si(³) and Si(6) were associated with high grain yield, but the other nonparametric stability measures were not positively correlated with grain yield and instead characterized a static concept of stability. Choice of genotype for increased grain yield in bread wheat would, therefore, be expected to change yield stability by increasing the values of the Si(³), Si(6) and TOP nonparametric stability statistics.
  • Allele frequency and selection efficiency in cross populations of Andean x Mesoamerican common beans (Phaseolus vulgaris L. Fabales, Fabaceae) Plant Genetics

    Baldoni, Aisy B.; Ramalho, Magno A. Patto; Abreu, Ângela de Fátima B.

    Resumo em Inglês:

    Strategies were investigated for improving efficiency in the use of segregating common bean (Phaseolus vulgaris) populations using crosses between the Andean cultivar BRS-Radiante and the Mesoamerican parent cultivar Carioca-MG by developing populations with 12.5%, 25%, 50%, 75% and 87.5% of the allele frequency of one of the parents. For each of the five populations we evaluated for two traits, the number of days to the beginning of flowering and grain yield (g plot-1), in the F2:3 (sown in February 2006) and F2:4 (sown in July 2006) generation progenies using 15 x 15 lattice design experiments, with 44 progenies (n = 220 plants) plus the two parents and three controls being evaluated for each generation. In terms of variability release, the populations with different parental allele frequencies presented no consistent tendency of alteration. In general, genetic variance was stated among progenies in all populations, indicating success with selection. For grain yield, the lowest mean was observed in the populations with 50% of the alleles of both parents. If, for instance, the objective is to develop earlier flowering lines, the best strategy is to perform two, or at least one, backcross with the earliest parent. The most suitable allele frequency is to be determined according to the desired grain type.
  • Evolutionary history of Phakopsora pachyrhizi (the Asian soybean rust) in Brazil based on nucleotide sequences of the internal transcribed spacer region of the nuclear ribosomal DNA Plant Genetics

    Freire, Maíra C. M.; Oliveira, Luiz O. de; Almeida, Álvaro M. R. de; Schuster, Ivan; Moreira, Maurilio A.; Liebenberg, Merion M.; Mienie, Charlotte M. S.

    Resumo em Inglês:

    Phakopsora pachyrhizi has dispersed globally and brought severe economic losses to soybean growers. The fungus has been established in Brazil since 2002 and is found nationwide. To gather information on the temporal and spatial patterns of genetic variation in P. pachyrhizi , we sequenced the nuclear internal transcribed spacer regions (ITS1 and ITS2). Total genomic DNA was extracted using either lyophilized urediniospores or lesions removed from infected leaves sampled from 26 soybean fields in Brazil and one field in South Africa. Cloning prior to sequencing was necessary because direct sequencing of PCR amplicons gave partially unreadable electrophoretograms with peak displacements suggestive of multiple sequences with length polymorphism. Sequences were determined from four clones per field. ITS sequences from African or Asian isolates available from the GenBank were included in the analyses. Independent sequence alignments of the ITS1 and ITS2 datasets identified 27 and 19 ribotypes, respectively. Molecular phylogeographic analyses revealed that ribotypes of widespread distribution in Brazil displayed characteristics of ancestrality and were shared with Africa and Asia, while ribotypes of rare occurrence in Brazil were indigenous. The results suggest P. pachyrhizi found in Brazil as originating from multiple, independent long-distance dispersal events.
  • Early colonization pattern of maize (Zea mays L. Poales, Poaceae) roots by Herbaspirillum seropedicae (Burkholderiales, Oxalobacteraceae) Plant Genetics

    Monteiro, Rose A.; Schmidt, Maria A.; Baura, Valter A. de; Balsanelli, Eduardo; Wassem, Roseli; Yates, Marshall G.; Randi, Marco A.F.; Pedrosa, Fábio O.; Souza, Emanuel M. de

    Resumo em Inglês:

    The bacterium Herbaspirillum seropedicae is an endophytic diazotroph found in several plants, including economically important poaceous species. However, the mechanisms involved in the interaction between H. seropedicae and these plants are not completely characterized. We investigated the attachment of Herbaspirillum to maize roots and the invasion of the roots by this bacterium using H. seropedicae strain SMR1 transformed with the suicide plasmid pUTKandsRed, which carries a mini-Tn5 transposon containing the gene for the Discosoma red fluorescent protein (Dsred) constitutively expressed together with the kanamycin resistance gene. Integration of the mini-Tn5 into the bacterial chromosome yielded the mutant H. seropedicae strain RAM4 which was capable of expressing Dsred and could be observed on and inside fresh maize root samples. Confocal microscopy of maize roots inoculated with H. seropedicae three days after germination showed that H. seropedicae cell were attached to the root surface 30 min after inoculation, were visible in the internal tissues after twenty-four hours and in the endodermis, the central cylinder and xylem after three days.
  • Molecular variability in the maize grey leaf spot pathogens in Brazil Plant Genetics

    Brunelli, Kátia R.; Dunkle, Larry D.; Sobrinho, Cândido A.; Fazza, Ana C.; Camargo, Luis E. A.

    Resumo em Inglês:

    Isolates of Cercospora species from leaves displaying symptoms of grey leaf spot were collected in maize-producing areas of south-central Brazil in 2001 and 2002. Restriction digests of the internal transcribed spacer region of rDNA detected the presence of the same two Cercospora species described on maize in the United States, namely C. zeae-maydis and the recently described species, C. zeina . Genetic variability among isolates was assessed by analysing 104 amplified fragment length polymorphism loci. Cluster analysis confirmed the genetic separation of isolates into two species with a mean similarity of 35%. Similarity levels within species were high, averaging 93% and 92% among isolates of C. zeae-maydis and C. zeina , respectively. The mean genetic similarity between C. zeae-maydis and C. zeina and two isolates of C. sorghi f. sp. maydis was 45% and 35%, respectively. Results of this study showed that populations of the grey leaf spot pathogens in Brazil are similar to those in the United States regarding species composition and that C. zeina is also present in Brazil.
  • Molecular characterization of Chaetomium species using URP-PCR Genetics Of Microorganisms

    Aggarwal, Rashmi; Sharma, Vandana; Kharbikar, Lalit L.; Renu,

    Resumo em Inglês:

    Chaetomium spp. are common colonizers of soil and cellulose-containing substrates. Seventeen isolates of Chaetomium spp., which included 15 isolates of C. globosum and one each of C. reflexum and C. perlucidum , were genetically characterized with universal rice primers (URP - primers derived from DNA repeat sequences in the rice genome) using polymerase chain reaction (URP-PCR). Out of the 12 URP's used in the study, nine primers were effective in producing polymorphic fingerprint patterns from DNA of Chaetomium spp . Analysis of the entire fingerprint profile using the unweighted pair-group method with arithmetic averages (UPGMA) clearly differentiated C. globosum isolates from C. perlucidum and C. reflexum . One of the primers, URP-2R, produced a uniform DNA band of 1.9 kb in all the isolates of C. globosum but not in C. perlucidum and C. reflexum , which can be used as molecular marker to differentiate C. globosum from other species. Our results indicate that URP's are sensitive and give reproducible results for assaying the genetic variability in Chaetomium spp.
  • Protection by Panax ginseng C.A. Meyer against the genotoxicity of doxorubicin in somatic cells of Drosophila melanogaster Mutagenesis

    Pereira, Denise G.; Antunes, Lusânia M.G.; Graf, Ulrich; Spanó, Mário A.

    Resumo em Inglês:

    Panax ginseng is one of the most widely prescribed herbal medicines for the treatment of cancer, diabetes, chronic inflammation, and neurodegenerative and cardiovascular diseases. Since the use of alternative medicines in combination with conventional therapy may increase the risk of unwanted interactions, we investigated the possible genotoxicity of a water-soluble form of the dry root of P. ginseng (2.5, 5.0 or 10.0 mg/mL) and its ability to protect against the genotoxicity of doxorubicin (DOX; 0.125 mg/mL) by using the Drosophila melanogaster wing somatic mutation and recombination test (SMART) with standard and high-bioactivation crosses of flies. Panax ginseng was not genotoxic at the concentrations tested, whereas DOX-induced genotoxicity in marker-heterozygous flies resulted mainly from mitotic recombination. At low concentrations, P. ginseng had antirecombinogenic activity that was independent of the concentration of extract used. Recombination events may promote cancer, but little is known about the ability of P. ginseng to inhibit such recombination or modulate DNA repair mechanisms.
  • Antigenotoxic activity and antioxidant properties of organic and aqueous extracts of pequi fruit (Caryocar brasiliense Camb.) pulp Mutagenesis

    Miranda-Vilela, Ana L.; Resck, Inês S.; Grisolia, Cesar K.

    Resumo em Inglês:

    The daily consumption of natural antioxidants protects against oxidative damage caused by reactive oxygen species (ROS), including DNA damage, and can reduce the risk of cancer, atherosclerosis and other degenerative diseases. The pulp of pequi (Caryocar brasiliense Camb.) fruit, a tree native to the Brazilian savannah, contains several compounds with antioxidant properties, including carotenoids, vitamin C, phenolic compounds such as flavonoids, saponins and tannins, and essential oils. In this work, we examined the ability of organic and aqueous extracts of pequi fruit pulp to protect against the genotoxicity induced by two antineoplastic drugs, cyclophosphamide (CP) and bleomycin (BLM). Micronucleus tests with mouse bone marrow cells and single-cell gel electrophoresis (comet assay) with peripheral blood leukocytes were used to examine the effects of CP and BLM, respectively. The antioxidant activity of the extracts was assessed by measuring lipid peroxidation with the TBARS method in mouse plasma. The fruit pulp extracts had no clastogenic or genotoxic effects in the cells studied, but both extracts protected against oxidative DNA damage caused by BLM or CP, indicating an ability to inhibit chemical mutagenesis in vivo. However, the protective effect against oxidative DNA damage depended on the dose of extract used. At the doses tested, the aqueous extract enhanced lipid peroxidation in mice of both sexes, especially in males. In contrast, the organic extract enhanced lipid peroxidation only in male mice, with no significant effect in females. These results suggest that, with adequate adjustment of the dose, an organic extract of pequi fruit pulp could be a useful dietary supplement with natural antioxidant activity, at least in females.
  • The historical biogeography of Pteroglossus aracaris (Aves, Piciformes, Ramphastidae) based on Bayesian analysis of mitochondrial DNA sequences Evolutionary Genetics

    Pereira, Sérgio L.; Wajntal, Anita

    Resumo em Inglês:

    Most Neotropical birds, including Pteroglossus aracaris, do not have an adequate fossil record to be used as time constraints in molecular dating. Hence, the evolutionary timeframe of the avian biota can only be inferred using alternative time constraints. We applied a Bayesian relaxed clock approach to propose an alternative interpretation for the historical biogeography of Pteroglossus based on mitochondrial DNA sequences, using different combinations of outgroups and time constraints obtained from outgroup fossils, vicariant barriers and molecular time estimates. The results indicated that outgroup choice has little effect on the Bayesian posterior distribution of divergence times within Pteroglossus , that geological and molecular time constraints seem equally suitable to estimate the Bayesian posterior distribution of divergence times for Pteroglossus , and that the fossil record alone overestimates divergence times within the fossil-lacking ingroup. The Bayesian estimates of divergence times suggest that the radiation of Pteroglossus occurred from the Late Miocene to the Pliocene (three times older than estimated by the “standard” mitochondrial rate of 2% sequence divergence per million years), likely triggered by Andean uplift, multiple episodes of marine transgressions in South America, and formation of present-day river basins. The time estimates are in agreement with other Neotropical taxa with similar geographic distributions.
  • Ant-Based Phylogenetic Reconstruction (ABPR): A new distance algorithm for phylogenetic estimation based on ant colony optimization Evolutionary Genetics

    Vittori, Karla; Delbem, Alexandre C.B.; Pereira, Sérgio L.

    Resumo em Inglês:

    We propose a new distance algorithm for phylogenetic estimation based on Ant Colony Optimization (ACO), named Ant-Based Phylogenetic Reconstruction (ABPR). ABPR joins two taxa iteratively based on evolutionary distance among sequences, while also accounting for the quality of the phylogenetic tree built according to the total length of the tree. Similar to optimization algorithms for phylogenetic estimation, the algorithm allows exploration of a larger set of nearly optimal solutions. We applied the algorithm to four empirical data sets of mitochondrial DNA ranging from 12 to 186 sequences, and from 898 to 16,608 base pairs, and covering taxonomic levels from populations to orders. We show that ABPR performs better than the commonly used Neighbor-Joining algorithm, except when sequences are too closely related (e.g., population-level sequences). The phylogenetic relationships recovered at and above species level by ABPR agree with conventional views. However, like other algorithms of phylogenetic estimation, the proposed algorithm failed to recover expected relationships when distances are too similar or when rates of evolution are very variable, leading to the problem of long-branch attraction. ABPR, as well as other ACO-based algorithms, is emerging as a fast and accurate alternative method of phylogenetic estimation for large data sets.
  • Patterns of ribosomal DNA distribution in hylid frogs from the Hypsiboas faber and H. semilineatus species groups Evolutionary Genetics

    Nunes, Rosana dos Reis Abrante; Fagundes, Valéria

    Resumo em Inglês:

    We performed a comparative analysis of the nucleolus organizer region (NOR) distribution in the karyotypes of hylid frogs from two different taxonomic groups, Hypsiboas faber and H. semilineatus. Silver nitrate staining of NORs (Ag-NORs) and fluorescence in situ hybridization (FISH) with a rDNA probe were used to investigate the chromosomal location of rDNA loci in two species. The karyotype of H. semilineatus and the Ag-NORs distribution of the four species are presented herein for the first time. After conventional staining, the four species presented very similar karyotypes with 2n = 24, but Ag-NORs analyses revealed species-specific characteristics. H. albomarginatus, H. faber and H. semilineatus had one pair of interstitial Ag-NORs in the short arm of pairs 2, 11, and in the long arm of pair 7, respectively. H. pardalis presented telomeric NORs in the long arm of pair 11. Ag-NORs were heteromorphic in three of the species ( H. pardalis , H. semilineatus and H. albomarginatus ) and FISH confirmed the differential activity of rDNA genes in H. semilineatus . In the present study, 2n = 24 karyotypes could be distinguished by their Ag-NORs distribution. Our results further the knowledge about the cytogenetics of hylids from Brazil.
  • Genetic characterisation of populations of the critically endangered Goliath grouper ( Epinephelus itajara, Serranidae) from the Northern Brazilian coast through analyses of mtDNA Evolutionary Genetics

    Silva-Oliveira, Gláucia C.; Rêgo, Péricles S. do; Schneider, Horacio; Sampaio, Iracilda; Vallinoto, Marcelo

    Resumo em Inglês:

    The Goliath grouper ( Epinephelus itajara ) is one of the most endangered species of fish of the subfamily Epinephelinae. Slow to develop and mature, and dependent on mangrove habitats for breeding, the species also suffers intense harvesting, which has reduced drastically in numbers in many areas. To contribute to the understanding of the characteristics of E. itajara populations, we conducted a molecular genetics study of the species, focusing on populations from the Northern Brazilian coast. The mtDNA control region (D-loop) of 116 individuals from five localities (Bragança, Ajuruteua, Parnaíba, Fortaleza and Natal) was analysed, and a sequence of 499 base pairs identified. Analyses of the sequences indicated that genetic variability was generally lower in E. itajara than in other endangered species of the genus. AMOVA found no significant grouping structure among the populations. Nested Clade Analysis revealed a significant association between genetic variability and geographic distribution among only three populations (Ajuruteua, Parnaíba and Natal). Genetic diversity was higher in populations from the Amazon region, which may be related to the better conservation of mangrove habitats in this area. Therefore, the present study could be used for the implementation of conservation and management measures in order to protect and consolidate these populations.
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