Abstract in English:The -1185A/G polymorphism in the 5'-regulatory region of the von Willebrand factor (VWF) gene was associated with VWF plasma levels in a normal population. This study was undertaken to evaluate whether there is a relationship between this polymorphism and type 1 von Willebrand disease (VWD), a disorder characterized by a quantitative deficiency of VWF. The association between this polymorphism and plasma VWF levels in normal Brazilian individuals was also analyzed. Control subjects (n = 460) and type 1 VWD patients (n = 41) were studied. Polymerase chain reaction (PCR) amplification of the 864-bp VWF promoter region followed by AccII restriction-digestion was used to identify the -1185A/G genotypes. The -1185G allele frequency was 57% in normal individuals and 63% in type 1 VWD patients, this difference was not significant (p = 0.29). No significant association was observed between -1185A/G genotypes and VWF plasma levels in normal individuals, although VWF levels were in the same direction as those reported by another study, with subjects carrying the G allele having the lower levels. These results suggest that -1185A/G polymorphism is not associated with the partial deficiency of VWF in type 1 VWD patients.
Abstract in English:We examined the genetic structure and the effects of a bottleneck in populations of the water rat Nectomys squamipes, a primary host of Schistosoma mansoni. Eight microsatellite loci were studied in 7 populations from the Sumidouro region of the Brazilian state of Rio de Janeiro. Our data, covering a four-year period during which a bottleneck occurred, revealed substantial variation (6-31 alleles per locus) and high levels of both observed (0.718-0.789) and expected (0.748-0.832) heterozygosity. Most populations were in Hardy-Weinberg equilibrium without linkage disequilibrium between loci. Overall average genetic differentiation between populations (estimated with the F ST (q) and R ST (r) analogues was 0.037 for q and 0.060 for r. There was significant allelic and genotypic differentiation between populations, especially in pairwise comparisons that included the most geographically isolated population. Direct migration estimates showed a low rate of migration, indicating that infected N. squamipes populations had a limited ability to spread S. mansoni. When the pre- and post-bottleneck populations were compared there was no detectable reduction in heterozygosity or allele number, although a significant excess of heterozygosity was detected in the post-bottleneck population.
Abstract in English:Partial cytochrome b and 12S rDNA mitochondrial DNA sequences of eight representatives of the Ramphastidae family were analyzed. We applied the linearized tree method to identify sequences evolving at similar rates and estimated the divergence times among some of the taxa analyzed. After excluding Ramphastos tucanus and Capito dayi from our data set, the remaining taxa presented a constant rate of DNA substitution, and branch lengths could be re-estimated with a clock constraint using the maximum likelihood method. Branch lengths were calibrated assuming that Galliformes and Piciformes split around 100 million years ago (mya). Our results indicate that Ramphastinae, and probably Capitoninae, diverged from other Piciformes in the Late Cretaceous (~82 mya), suggesting that Piciformes is another avian order that survived the mass extinction event occurred 65 mya at the Cretaceous/Tertiary (K/T) boundary. The divergence times estimated within the Ramphastinae genera cover the period from the Middle Eocene (around 47 mya) through the Late Miocene (9.5 mya). Our estimate of divergence time is coincidental with the split of the African and the South American continents and other intense geologic activities and modifications of the areas which correspond to the current Neotropics. These events might have influenced the diversification of Ramphastinae in South America.
Abstract in English:Caffeine and used coffee grounds completely blocked the development of Aedes aegypti in the early stages, in treatments with the concentrations 1.0 mg/mL and 50 mg/mL, respectively. More advanced stages and even adults were obtained in lower concentrations of both substances, enabling observations to be made of mortality rate, longevity and esterase patterns. The experiments involved treatments using either eggs or 3rd instar larvae (L3), with or without the addition of fish food. Mortality rates prior to the adult stage and adult longevity were significantly different in the comparisons among treatments, in every kind of experiment, but in those using L3 larvae, their percentages were smaller. Observations of the time of larva and adult onset suggested that developmental time was also delayed in treatments with both substances. The addition of fish food increased significantly the number of adults produced in caffeine 0.2 and in the control, but in used coffee grounds, the opposite effect occurred. Longevity was apparently not affected by the addition of food, except again in coffee grounds, in which it decreased. In an attempt to detect a mechanism involved in the action of caffeine and coffee grounds, esterases (enzymes involved in the detoxification of xenobiotics) were analyzed in polyacrylamide gels of treated 4th instar larvae (L4). In treatments with both substances, the expression of some carboxylesterases was affected, suggesting that they may be involved in the observed impairment.
Abstract in English:Allelic frequencies at 12 loci (five blood groups: C, D, K, P, and U; and seven protein polymorphisms: Al, A1B, Es, Gc, Hb, PGD, and Tf), are given for two Brazilian horse breeds: Mangalarga Marchador and Mangalarga. The high genetic identity value found (96.0%) is consistent with their common origin, although, at some point of the development of Mangalarga Marchador, Mangalarga separated from the original stock. The expected average heterozygosity was higher in Mangalarga Marchador. The populations presented genetic differentiation, as shown by the statistically significant value of F ST. The nonsignificant F IS values showed that there was no appreciable consanguineous mating in any of the two populations. Exclusion probability calculated for the 12 loci was 87.0% and 86.5% for Mangalarga Marchador and Mangalarga, respectively. No genetic equilibrium was observed in the A1B, Tf, and Es loci of Mangalarga Marchador. The frequencies of blood factors A, Q, and T were calculated.
Abstract in English:The specific adaptation of 15 rubber tree (Hevea brasiliensis) clones was assessed by analyzing yield during a normal year (1997-98) and a year (1998-99) in which the yield was exceptional. Differences in yield in response to changes in weather conditions over the years were evident with clones RRII 203, RRIM 703, PB 5/51 and PB 235 which all exhibited a negative trend with increasing wind velocity during 1997-98, these clones also exhibited a negative correlation with minimum temperature during 1998-99. The prominent yield differences across the years made selection based on both yield and stability inevitable through computing weather variables and environmental index as covariant. To determine the contribution of variable(s) to genotype-environment (GE) interactions, the GE interaction was partitioned into heterogeneity and residual GE interaction. Heterogeneity only for environmental index was highly significant (p = 0.01), meaning that stability or instability of clones was due to a linear effect of the environmental index. The non-significant values of heterogeneity for the weather variables revealed that none of these factors individually was sufficient to explain heterogeneity. A QBASIC computer program called STABLE was used to select simultaneously for yield and stability. Clones PB 235, RRII 118, RRII 203, RRIM 703 and RRIM 600 were stable over the years investigated.
Abstract in English:The best-yielding, best vigour and most stable Hevea clones are identified by growing clones in different environments. However, research on the stability in Hevea brasiliensis (Willd. Adr. ex Juss.) Muell.-Arg. is scarce. The objectives of this work were to assess genotype-environment interaction and determine stable genotypes. Stability analysis were performed on results for girth growth and rubber yield of seven clones from five comparative trials conducted over 10 years (girth growth) and four years (rubber yield) in São Paulo State, Brazil. Stability was estimated using the Eberhart and Russell (1966) method. Year by location and location variability were the dominant sources of interactions. The stability analysis identified GT 1 and IAN 873 as the most stable clones for girth growth and rubber yield respectively since their regression coefficients were almost the unity (b = 1) and they had one of the lowest deviations from regressions (S2di). Their coefficient of determination (R²) was as high as 89.5% and 89.8% confirming their stability. In contrast, clones such as PB 235, PR 261, and RRIM 701 for girth growth and clones such as GT 1 for rubber yield with regression coefficients greater than one were regarded as sensitive to environment changes.
Abstract in English:The ''cagaita tree'' (Eugenia dysenterica) is a plant found widespread in the Brazilian Cerrado. Its fruit is used for popular consumption and for industrial purposes. This study opens a new perspective for the generation of population genetic data and parameters estimates for devising sound collection and conservation procedures for Eugenia dysenterica. A battery of 356 primer pairs developed for Eucalyptus spp. was tested on the ''cagaita tree''. Only 10 primer pairs were found to be transferable between the two species. Using a polyacrilamide gel, an average of 10.4 alleles per locus was detected, in a sample of 116 individuals from 10 natural ''cagaita tree'' populations. Seven polymorphic loci allowed estimation of genetic parameters, including expected average heterozygosity He = 0,442, among population diversity, R ST = 0,268 and gene flow Nm = 0,680. Results indicated a potential of SSR locus transferability developed for Eucalyptus to other species of different genera, such as in the case of the ''cagaita tree''. The high genetic diversity among populations detected with SSR markers indicated that these markers are highly sensitive to detect population structure. Estimated Nm values and the existence of private alleles indicated reduced gene flow and consequently possible damage to the metapopulation structure.
Abstract in English:The objective of this study was to identify RAPD and SSR markers associated with a resistant allele for angular leaf spot (Phaeoisariopsis griseola) from the line 'ESAL 550', derived from the Andean 'Jalo EEP 558' cultivar, to assist selection of resistant genotypes. The resistant line 'ESAL 550' and the susceptible cultivar 'Carioca MG' were crossed to generate F1 and F2 populations. One hundred and twenty F2:3 families were evaluated. The DNA of the 12 most resistant families was bulked and the same was done with the DNA of the 10 most susceptible, generating two contrasting bulks. One RAPD and one SSR marker was found to be linked in coupling phase to the resistant allele. The SSR marker was amplified by the primer PV-atct001(282C), and its distance from the resistant allele was 7.6 cM. This is the most useful marker for indirect selection of resistant plants in segregating populations. The RAPD marker was amplified by the primer OPP07(857C) linked in coupling phase to the resistant allele, and distant 24.4 cM. Therefore, this RAPD marker is not so useful in assisting selection because it is too far from the resistant allele.
Abstract in English:Estimation of general and specific combining ability effects in a diallel analysis of cross-pollinating populations, including the selfed parents, is presented in this work. The restrictions considered satisfy the parametric values of the GCA and SCA effects. The method is extended to self-pollinating populations (suitable for other species, without the selfed parents). The analysis of changes in population means due to inbreeding (sensitivity to inbreeding) also permits to assess the predominant direction of dominance deviations and the relative genetic variability in each parent population. The methodology was used to select popcorn populations for intra- and inter-population breeding programs and for hybrid production, developed at the Federal University of Viçosa, MG, Brazil. Two yellow pearl grain popcorn populations were selected.
Abstract in English:Rice blast is the most important fungal disease of rice and is caused by Pyricularia oryzae Sacc. (Telomorph Magnoporthe grisea Barr.). Seven randomly amplified polymorphic DNA (RAPD) markers OPA5, OPG17, OPG18, OPG19, OPF9, OPF17 and OPF19 showed very clear polymorphism in resistant cultivar lines which differed from susceptible lines. By comparing different susceptible lines, nine DNA amplifications of seven primers (OPA5(1000), OPA5(1200,) OPG17(700), OPG18(850), OPG19(500), OPG19(600), OPF9(600), OPF17(1200) and OPF19(600)) were identified as dominant markers for the blast resistant gene in resistant cultivar lines. These loci facilitate the indirect scoring of blast resistant and blast susceptible genotypes. The codomine RAPDs markers will facilitate marker-assisted selection of the blast resistant gene in two blast resistant genotypes of rice (Labelle and Line 11) and will be useful in rice breeding programs.
Abstract in English:Iron is essential for plants. However, excess iron is toxic, leading to oxidative stress and decreased productivity. Therefore, plants must use finely tuned mechanisms to keep iron homeostasis in each of their organs, tissues, cells and organelles. A few of the genes involved in iron homeostasis in plants have been identified recently, and we used some of their protein sequences as queries to look for corresponding genes in the rice (Oryza sativa) genome. We have assigned possible functions to thirty-nine new rice genes. Together with four previously reported sequences, we analyzed a total of forty-three genes belonging to five known protein families: eighteen YS (Yellow Stripe), two FRO (Fe3+-chelate reductase oxidase), thirteen ZIP (Zinc regulated transporter / Iron regulated transporter Protein), eight NRAMP (Natural Resistance - Associated Macrophage Protein), and two Ferritin proteins. The possible cellular localization and number of potential transmembrane domains were evaluated, and phylogenetic analysis performed for each gene family. Annotation of genomic sequences was performed. The presence and number of homologues in each gene family in rice and Arabidopsis is discussed in light of the established iron acquisition strategies used by each one of these two plants.
Abstract in English:Paspalum quadrifarium Lam. is a bunchgrass native to Uruguay, Argentina, and southern Brazil. Diploid, triploid, tetraploid and hexaploid cytotypes have been reported for this species of the Quadrifaria group of Paspalum. In this group, a high degree of cytogenetic homology between the genomes of several diploid species has been reported, based on meiotic pairing in interspecific hybrids; multivalent associations would thus be expected in polyploid hybrids. Karyotype analysis could provide useful information about the genomic architecture of polyploid plants; however, the fully condensed mitotic chromosomes of Paspalum do not provide enough morphological features for such an analysis. In this paper, we used mitotic prometaphase chromosomes treated with 70% acetic acid at 40 °C after cover slip removal. This process removes cytoplasm that remains from chromosome squashes and makes prometaphases available for karyological analysis. The karyotypes of a triploid (2n = 3x = 30) and a tetraploid (2n = 4x = 40) accession of Paspalum quadrifarium were studied using this technique, and evidence of segmental allopolyploidy was found in both cases. In both accessions, meiotic behavior was in accordance with that origin. This technique greatly improved the number and quality of analyzable metaphases and prometaphases on otherwise conventional slides and is recommended for plants with small chromosomes.
Abstract in English:Thinopyrum ponticum (2n = 10x = 70, JJJJsJs) belongs to the Triticeae tribe, and is currently used as a source of pathogen resistance genes in wheat breeding. In order to characterize its chromosomes, the number and position of 45S and 5S rDNA sites, as well as the distribution of the repetitive DNA sequences pAs1 and pSc119.2, were identified by fluorescent in situ hybridization. The number of nucleoli and NORs was also recorded after silver nitrate staining. Seventeen 45S and twenty 5S rDNA sites were observed on the short arms of 17 chromosomes, the 45S rDNA was always located terminally. On three other chromosomes, only the 5S rDNA site was observed. Silver staining revealed a high number of Ag-NORs (14 to 17) on metaphase chromosomes, whereas on interphase nuclei there was a large variation in number of nucleoli (one to 15), most of them (82.8%) ranging between four and nine. The pAs1 probe hybridized to the terminal region of both arms of all 70 chromosomes. In addition, a disperse labeling was observed throughout the chromosomes, except in centromeric and most pericentromeric regions. When the pSc119.2 sequence was used as a probe, terminal labeling was observed on the short arms of 17 chromosomes and on the long arms of five others. The relative position of 45S and 5S rDNA sites, together with the hybridization pattern of pAs1 and pSc119.2 probes, should allow whole chromosomes or chromosome segments of Th. ponticum to be identified in inbred lines of wheat x Th. ponticum.
Abstract in English:In this study, we identified disease resistance gene homologs in Brassica oleracea and assessed their expression in lines resistant and susceptible to Xanthomonas campestris pv. campestris (Xcc). Two DNA fragments of approximately 2.5 kb (BI-16/RPS2 and Lc201/RPS2) were amplified by PCR from two Brassica lines using primers based on an RPS2 homologous sequence previously described in the Brassica oleracea ecotype B117. The sequences of these fragments shared high similarity (95-98%) with RPS2 homologs from various Brassica species. The digestion of these fragments with restriction enzymes revealed polymorphisms at the Xba I restriction sites. The length polymorphisms were used as a co-dominant marker in an F2 population developed to segregate for resistance to Xcc, the causal agent of black rot. Linkage analysis showed no significant association between the marker and quantitative trait loci for black rot. RT-PCR with specific primers yielded an expected 453 bp fragment that corresponded to the RPS2 homologs in both resistant and susceptible lines inoculated with the pathogen, as well as in non-inoculated control plants. These results suggest that these homologs are constitutively expressed in B. oleracea.
Abstract in English:Cytogenetic studies were carried out on 22 accessions of Brachiaria jubata from the Embrapa Beef Cattle Brachiaria collection. One accession was diploid (2n = 2x = 18) and the remaining 21 were tetraploid (2n = 4x = 36). Among five tetraploid accessions, a specific and constant pattern of cell fusion involving only two microsporocytes was recorded. Meiosis proceeded normally from prophase I to the end, giving rise to an octad with normal microspores that developed into fertile pollen grains. Regular octad formation was possible because each cellular chromosome set was maintained in its proper domain, spindles were correctly positioned, and cytokinesis planes were formed in the correct places. Such behavior of meiosis in syncytes has never been reported in any other plant species.
Abstract in English:Hansenula polymorpha is a methylotrophic yeast widely employed in biotechnology as a ''protein factory''. Most promoters used for heterologous protein expression, like MOX (methanol oxidase) and DAS (di-hydroxy acetone synthase), are involved in the peroxisomal methanol metabolism (C1 metabolism) and are under strong glucose repression. Interestingly, the MOX promoter is subjected to glucose regulation also in Saccharomyces cerevisiae, a non-methylotrophic yeast in which this phenomenon is well studied. In this species, the transcription factor Tup1p plays an essential role in glucose repression of several genes. This effect is counteracted by the activator Snf1p when glucose is exhausted from medium. Therefore, to test whether this regulatory circuit has been conserved in H. polymorpha, HpTUP1 and HpSNF1 were partially cloned and disrupted. Deletion of HpTUP1 did not affect glucose repression of the major C1 metabolism genes (MOX, DAS). Thus, though conserved, HpTUP1 does not seem to take part in a general glucose repression in H. polymorpha. In contrast, the deletion of HpSNF1 led to significant decreases in the activation of these genes in the absence of glucose. Therefore, the effect of HpSNF1 in transcriptional activation may be through an HpTUP1- independent circuit.
Abstract in English:Dependencies in DNA sequences are frequently modeled using Markov models. However, Markov chains cannot account for heterogeneity that may be present in different regions of the same DNA sequence. Hidden Markov models are more realistic than Markov models since they allow for the identification of heterogeneous regions of a DNA sequence. In this study we present an application of hidden Markov models to a subsequence of the Xylella fastidiosa DNA data. We found that a three-state model provides a good description for the data considered.
Abstract in English:Herbaspirillum spp. are endophytic diazotrophic bacteria associated with important agricultural crops. In this work, we analyzed six strains of H. seropedicae (Z78, M2, ZA69, ZA95, Z152, and Z67) and one strain of H. rubrisubalbicans (M4) by restriction fragment length polymorphism (RFLP) using HindIII or DraI restriction endonucleases, random amplified polymorphic DNA (RAPD), and partial sequencing of 16S rDNA. The results of these analyses ascribed the strains studied to three distinct groups: group I, consisting of M2 and M4; group II, of ZA69; and group III, of ZA95, Z78, Z67, and Z152. RAPD fingerprinting showed a higher variability than the other methods, and each strain had a unique electrophoretic pattern with five of the six primers used. Interestingly, H. seropedicae M2 was found by all analyses to be genetically very close to H. rubrisubalbicans M4. Our results show that RAPD can distinguish between all Herbaspirillum strains tested.
Abstract in English:During the last few decades the search for medical treatments based on alternative medicine has increased significantly, making knowledge of the plants commonly used as folk medicines extremely important. The plant Cochlospermum regium, a member of the Cochlospermaceae found in the Brazilian cerrado (a type of savanna), is known to have high depurative activity and to be effective not only in treating skin problems such as pimples, boils and blotches but also in curing gastritis and ulcers. We prepared aqueous extracts using 13, 19 and 25 gL-1 of dried C. regium root and investigated these extracts for possible mutagenic effects on Drosophila melanogaster germ cells. Mutagenesis was assessed using the ring-X loss (RXL) test which can detect chromosome mosaicism, partial loss of the ring X chromosome and chromosome non-disjunction. Our results showed that at the concentrations tested C. regium extracts did not induce ring-X loss in D. melanogaster.
Abstract in English:The use of medicinal plants by the general population is an old and still widespread practice, which makes studies of their genotoxicity essential. Psidium guajava L. and Achillea millefolium L. are examples of plants commonly used in popular medicine. P. guajava L. is indicated for diarrhea and also as an antiseptic, while A. millefolium L. is indicated as an analgesic, antispasmodic, digestive, diuretic, antiseptic, astringent, emollient, wound healer and hemorrhoid medication. The aim of this study was to determine the effects of the infusions of these two plant species on chromosomes and the cell cycle. Leaves from the plants were used to prepare infusions, in the same manner as teas, but at two different concentrations. Allium cepa L. root-tip cells (P. guajava L. - 2.62 and 26.2 mg/mL, and A. millefolium L. - 3.5 and 35.0 mg/mL) and Wistar rat bone marrow cells (P. guajava L. - 2.62 and 26.2 mg/100g body weight, and A. millefolium L. - 3.5 and 35.0 mg/100g body weight) were used as in vivo plant and animal test systems, respectively. Human peripheral blood lymphocytes (P. guajava L. - 0.262 and 2.62 mg/mL culture medium, and A. millefolium L. - 0.35 and 3.5 mg/mL culture medium) were used as in vitro test system. The P. guajava L. infusion at the higher concentration caused a statistically significant inhibition of cellular division in the onion root-tip cells, not observed in onion root-tip cells treated with A. millefolium L. No statistically significant alterations were found, as compared to untreated controls, in either the cell cycle or the number of chromosome alterations, after treatments with either plant, in rat cells or in cultured human lymphocytes. These results regarding the cytotoxicity and mutagenicity of these plants provide valuable information about the safety of using them as therapeutic agents.