Identification of sex in Carica papaya L. using RAPD markers

Brazil is currently the worlds largest producer of papaya (Carica papaya L.), producing fruits for both the domestic market and export. Only fruits from hermaphrodite plants are marketed because they have the necessary commercial characteristics, i.e. they are pear-shaped and have thicker flesh and a smaller internal cavity. Increased papaya yield has been limited mainly by the ratio of female to hermaphrodite (1:2) plants normally occurring in orchards. This ratio causes great losses to papaya producers and the identification of the sex of seedlings during the nursery stage would be an important advance. In our study random amplified polymorphic DNA (RAPD) analysis was used to differentiate between the sexual forms of three commercial C. papaya cultivars belonging to the Solo group. RAPD assays using the BC210 primer were able to detect hermaphrodites in all of the cultivars tested. The BC210(438)molecular marker was much better at papaya sex differentiation than other markers described in the literature.

Genetic variability in species of bats revealed by RAPD analysis

Random amplified polymorphic DNA molecular marker was utilized as a means of analyzing genetic variability in seven bat species: Molossus molossus, M. rufus, Eumops glaucinus, E. perotis, Myotis nigricans, Eptesicus furinalis, and Artibeus planirostris. The determination of genetic diversity was based on 741 bands produced by a 20-random primer set. Only eight bands were considered monomorphic to one species. The greatest number of bands and the most polymorphic condition were exhibited by M. molossus, followed by M. nigricans, A. planirostris, E. furinalis, E. glaucinus, M. rufus, and E. perotis. Nei's genetic diversity index in the seven species considering the 20 primers was not greater than 0.22, but some primers were capable of detecting values between 0.39 and 0.49. Nei's unbiased genetic distance values and the UPGMA clustering pattern show that M. molossus and M. rufus have a close genetic relationship, unlike that observed between E. perotis and E. glaucinus. The latter was clustered with A. planirostris and E. furinalis. The low values for genetic diversity and distance observed indicate a genetic conservatism in the seven species. The fluorescent in situ hybridization experiments did not confirm a monomorphic condition for the eight bands identified, demonstrating that the monomorphic bands obtained by random amplified polymorphic DNA are insufficient for the identification of bat species.

RAPD genomic fingerprinting differentiates Thiobacillus ferrooxidans strains

The PCR-based technique, involving the random amplification of polymorphic DNA (RAPD), was optimized and used for assessing genomic variability among eight Thiobacillus ferrooxidans strains. RAPD fingerprints presented variation for the thirty primers used, giving a total of 269 polymorphic bands. Similarity coefficients between the strains were calculated, and UPGMA cluster analysis was used to generate a dendrogram showing relationships among them. Most primers divided T. ferrooxidans strains in two distinct groups - Group 1: S, SSP, V3, AMF and Group 2: CMV, FG-460, I-35, LR. We observed that the T. ferrooxidans strains used in this work have a high degree of genomic diversity and that RAPD is a powerful method to differentiate them.

Evaluation of the Spreading of Isolated Bacterias from Dental Consulting-Room using RAPD Technique

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diversidade genética de quatro linhagens de Oreochromis niloticus utilizando o marcador RAPD

The aim of this study was to analyze the genetic diversity of four Nile Tilapia (Oreochromis niloticus) strains using the RAPD marker. Fin samples of GIFT (G), Chitralada (C), Supreme (S) and Bouake (B) juvenile stocks have been collected. The 11 primers used yielded 81 fragments of which 41.98% were polymorphic. The percentage of polymorphic loci (G: 18.52%; C: 19.75%; S: 20.99% and B: 24.79%) showed that there was a genetic differentiation among the strains, showing the G(st) values a high (BxG: 0.231; BxC: 0.224; GxC: 0.194 and SxC: 0.208) and elevated (BxS: 0.315 and GxS: 0.270) differentiation. The highest gene flow (N(m)) was among the GxC (2.082) strains. The distance and genetic identity values (0.044 and 0.957 respectively) and the dendrogram indicate that the GxC is the most genetically similar strains. The genetic similarity was high among of the strains (G: 0,932; C: 0,903; S: 0,891 and B: 0.900). These results will enable a correct reproductive and genetic strains management.

Genetic variation and phylogenetic relationships based on RAPD analysis in section Caulorrhizae, genus Arachis (Leguminosae)

Wild Arachis germplasm includes potential forage species, such as the rhizomatous Arachis glabrata and the stoloniferous A. pinto and A. repens. Commercial cultivars of A. pintoi have already been released in Australia and in several Latin American countries, and most of these cultivars were derived from a single accession of A. pintoi (GK 12787). Arachis repens is less productive as a forage plant than is A. pintoi. However, it can be crossed with A. pintoi, and thus has good potential as germplasm for the improvement of A. pintoi. Arachis repens is also used as an ornamental plant and ground cover. Many new accessions of these two stoloniferous species are now available, and they harbor significant genetic variability beyond that available in the few older accessions, previously available. Therefore, these new accessions need to be conserved, documented and considered in terms of their potential for crop improvement and direct commercial use. Sixty-four accessions of this new germplasm were analyzed using RAPD analysis. Most of the accessions of A. repens grouped together into a clearly distinct group. In general, the accessions from the distinct valleys of the Jequitinhonha, Sao Francisco and Parana rivers did not group together, suggesting there is not a tight relation between dispersion by rivers and the geographic distribution of genetic variation in these species.

Avaliação da variabilidade genética em quatro gerações de Eucalyptus urophylla S.T. Blake por meio do marcador molecular RAPD

Molecular markers have gradually replaced morphological markers in population studies. The advantages of molecular markers are the speed and precision of evaluations, mainly for long cycle cultures, where determinate traits can take years to manifest. The principle objectives of this research were to assess variability and genetic distances in four generations of Eucalyptus urophylla and provide data that help with the continued improvement of these materials. The populations can be found at the Experimental Forestry Sciences Station, Anhembi, SP, belonging to the College of Agriculture Luiz de Queiroz of São Paulo University. The initial base population was introduced by seeds collected in indonesia and designated P0 generation. The subsequent segregated generations, derivatives of recombination starting with open pollination, were designated P1, P2, and P3. One hundred and seventy four individual trees representing the four generations were analysed. The RAPD technique allowed the identification of 86 loci that were analysed with the Jaccard Coefficient, generating a genetic similarity matrix, permitting the estimation of genetic distances. The genetic distance of generation PO was 0.3338333, P1 was 0.336824, P2 was 0.40000, and P3 was 0.381093. In percentage terms the genetic distances between individuals grew in relation to base population, being 0.15% for generation P1, 18.93% for P2, and 13.31% for P3. This shows an increase in genetic variability with the advance of the program, despite the selective processes. From this came the belief that the initial base population was resulting from seed collection from isolated trees. These populations, although going through successive selections, had a high cross efficiency through satisfactory pollination, which then permitted genetic variation to increase, the outcome of effective recombination between individuals. Generations P2 and P3 gave a better perspective for the continuance of the improvement program due to the high number of different groups with standard genetic distances of 35%. The selections made between the diverse genetic groups allowed the efficient use of genetic variability evaluation.

Caracterização silvicultural, botânica e avaliação da variabilidade genética por meio do marcador molecular RAPD em um teste de progênies de Eucalyptus urophylla S. T. Blake

Molecular markers have recently been incorporated into genetic improvement programs. They are already considered as powerful tools with several different uses, for instance the monitoring of genetic variability in tree populations. The main objectives of this study were to evaluate genetic variability in Eucalyptus urophylla progenies and together with silvicultural and botanical information, provide assistance to the improvement program. The Eucalypts population is located at the Experimental Forestry Sciences Station, Anhembi, SP, which belongs to the College of Agriculture Luiz de Queiroz. Sixty-nine progenies were analysed representing one individual by family in open pollinated Eucalyptus urophylla trees. The RAPD technique allowed the identification of 72 loci that were analysed using Jaccard's Coefficient generating a genetic similarity matrix to permit estimation of genetic distances. The results obtained showed genetic distance between individuals of 0.40 with 12 groups of genetic variability using a standardised distance of 40%. The progenies showed different bark patterns, allowing the establishment of bark groups. The groups formed based on genetic distances obtained using DNA analysis did not correspond to those based on bark pattern. Genetic selection was simulated in which silvicultural and genetic variability data were linked, thus avoiding excessive variability losses. The simulation of controlled crossings allowed the maximum genetic difference to be obtained linked with height and individual bark roughness.

RAPD analysis in the Neotropical fish Brycon lundii: Genetic diversity and its implications for the conservation of the species

Random amplified polymorphic DNA (RAPD) technique was used to examine the genetic variability on an endangered Neotropical fish species, Brycon lundii, collected on two regions with distinct environmental conditions in the São Francisco River (Brazil), downstream from a hydroelectric station. Using decamer oligonucleotides as single primers in Polymerase Chain Reaction (PCR), genetic similarity index, mean allele frequency and mean heterozigosity were estimated, revealing variations between samples from the two regions. Moreover, a fragment of about 1200 base pairs was found in 100% of the examined animals collected at the region closer to the hydroelectric dam, while its frequency was much lower (27.3%) within the sample from the second collecting site, 30 km downstream from the dam, indicating a possible correlation between genetic variation and geographical area. A dendogram representing the relationships among genotypes was obtained, demonstrating at least two major clusters of animals. Based on the data, a model of population structuring in Brycon lundii is suggested. The described approach holds great promise for further analyses and gives support to biodiversity maintenance and recovery efforts of B. lundii.

Evaluation of DNA polymorphisms amplified by arbitrary primers (RAPD) as genetically associated elements to differentiate virulent and non-virulent Paracoccidioides brasiliensis isolates

Randomly amplified polymorphic DNA (RAPD) analysis of 35 Paracoccidioides brasiliensis isolates was carried out to evaluate the correlation of RAPD profiles with the virulence degree or the type of the clinical manifestations of human paracoccidioidomycosis. The dendrogram presented two main groups sharing 64% genetic similarity. Group A included two isolates from patients with chronic paracoccidioidomycosis; group B comprised the following isolates showing 65% similarity: two non-virulent, six attenuated, five virulent, eight from patients with chronic paracoccidioidomycosis and two from patients with acute paracoccidioidomycosis. The virulent Pb18 isolate and six attenuated or non-virulent samples derived from it were genetically indistinguishable (100% of similarity). Thus, in our study, RAPD patterns could not discriminate among 35 P. brasiliensis isolates according to their differences either in the degree of virulence or in the type of the clinical manifestation of this fungal infection. © 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Genetic divergence in Eucalyptus spp. clones by molecular RAPD markers

The genetic divergence in 20 Eucalyptus spp. clones was evaluated by multivariate techniques based on 167 RAPD markers, of which 155 were polymorphic and 12 monomorphic. The measures of genetic distances were obtained by the arithmetic complement of the coefficients of Jaccard and of Sorenso-Nei and Li and evaluated by the hierarchical methods of Single Linkage clustering and Unweighted Pair Group Method with Arithmetic Mean (UPGMA). Independent of the dissimilarity coefficient, the greatest divergence was found between clones 7 and 17 and the smallest between the clones 11 and 14. Clone clustering was little influenced by the applied procedure so that, adopting the same percentage of divergence, the UPGMA identified two groups less for the coefficient of Sorenso-Nei and Li. The clones evidenced considerable genetic divergence, which is partly associated to the origin of the study material. The clusters formed by the UPGMA clustering algorithm associated to the arithmetic complement of Jaccard were most consistent.

RAPD optimization for genetic studies with Citrulus lanatus and Sesamum indicum genotypes

The Random Amplified Polymorphic DNA (RAPD) technique is powerful for DNA polymorphism determinations and is widely used in research involving different organisms, but it is known that RAPD can be affected by many factors that may result in false positive bands and non-reproducible assays. In this study, we analyzed the effect of several factors such as DNA template, primer and Taq DNA polymerase concentrations to optimize and standardize the RAPD technique for further genetic studies with Citrulus lanattus and Sesamum indicum L. The best combination of DNA, Taq DNA polymerase enzyme and primer concentrations in RAPD amplification procedures for sesame and watermelon genotypes was established.

Homeostase do cálcio e marcadores do metabolismo ósseo no hipertireoidismo felino - Revisão

Hyperthyroidism is the most frequent endocrine disease in old-aged cats. It is a illness provoked by the excess of circulating thyroid hormones. Hyperthyroidism causes alteration in bone metabolism with predominance of activity resorption. The evaluation of bone metabolism can be made by measuring serum and urinary markers of bone metabolism or bone mineral densitometry. Osteoblasts are responsible cells for bone formation while the osteoclasts are for resorption. In physiological situation osteoblastic and osteoclastic activities are in balance. Markers of bone formation express the osteoblastic activity and markers of the osseous resorption the osteoclástica activity. Markers of bone turnover are important in the diagnosis and prognostic of muscle-skeletal disease, as well as in the accompaniment of therapy. It is fundamental do carry on studies on the influence of feline hyperthyroidism on markers of bone formation and resorption in bone turnover to comprise pathophysiologic mechanism of bone alterations.