Caracterização de rizóbios indicados para produção de inoculantes por meio de sequenciamento parcial do 16S rRNA

O objetivo deste trabalho foi confrontar as sequências parciais do gene 16S rRNA de estirpes padrão de rizóbios com as de estirpes recomendadas para a produção de inoculantes no Brasil, com vistas à verificação da confiabilidade do sequenciamento parcial desse gene para a identificação rápida de estirpes. Foram realizados sequenciamentos através de reação em cadeia da polimerase (PCR) com iniciadores relativos à região codificadora do gene 16S rRNA entre as bactérias estudadas. Os resultados foram analisados pela consulta de similaridade de nucleotídeos aos do Basic Local Alignment Search Tool (Blastn) e por meio da interpretação de árvores filogenéticas geradas usando ferramentas de bioinformática. A classificação taxonômica das estirpes Semia recomendadas para inoculação de leguminosas com base em propriedades morfológicas e especificidade hospedeira não foi confirmada em todas as estirpes. A maioria das estirpes estudadas, consultadas no Blastn, é consistente com a classificação proposta pela construção de árvores filogenéticas das sequências destas estirpes, com base na similaridade pelo sequenciamento parcial do gene considerado.

Phylogeography and Pleistocene demographic history of the endangered marsh deer (Blastocerus dichotomus) from the Rio de la Plata Basin

The marsh deer is the largest neotropical cervid with morphological and ecological adaptations to wetlands and riparian habitats. Historically, this now endangered species occupied habitats along the major river basins in South America, ranging from southern Amazonia into northern Argentina to the Parana river delta. This particularly close association with wetlands makes marsh deer an excellent species for studying the effects of Pleistocene climatic changes on their demographic and phylogeographic patterns. We examined mitochondrial DNA variation in 127 marsh deer from 4 areas distributed throughout the Rio de]a Plata basin. We found 17 haplotypes in marsh deer from Brazil, Bolivia and Argentina that differed by 1-8 substitutions in a 601 bp fragment of mitochondrial control region sequence, and 486 bp of cytochrome b revealed only 3 variable sites that defined 4 haplotypes. Phylogeny and distribution of control region haplotypes suggest that populations close to the Pantanal area in central Brazil underwent a rapid population expansion and that this occurred approximately 28,000-25,000 years BP. Paleoclimatic data from this period suggests that there was a dramatic increase for precipitation in the medium latitudes in South America and these conditions may have fostered marsh deer's population growth.

Elucidating the Evolution of the Red Brocket Deer Mazama americana Complex (Artiodactyla; Cervidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influência da filogenia em redes de interações ecológicas

In this thesis we deal with a class of composed networks that are formed by two tree networks, TP and TA, whose end points touches each other through a bipartite network BPA. We explore this network using a functional approach. We are interested in what extend the topology, or the structure, of TX (X = A or P) determines the links of BPA. This composed structure is an useful model in evolutionary biology, where TP and TA are the phylogenetic trees of plants and animals that interact in an ecological community. We use in this thesis two cases of mutualist interactions: frugivory and pollinator networks. We analyse how the phylogeny of TX determines or is correlated with BPA using a Monte Carlo approach. We use the phylogenetic distance among elements that interact with a given species to construct an index κ that quantifies the influence of TX over BPA. The algorithm is based in the assumption that interaction matrices that follows a phylogeny of TX have a total phylogenetic distance smaller than the average distance of an ensemble of Monte Carlo realizations generated by an adequate shuffling data. We find that the phylogeny of animals species has an effect on the ecological matrix that is more marked than plant phylogeny

Cytogenetic characterization of the strongly electric Amazonian eel, Electrophorus electricus (Teleostei, Gymnotiformes), from the Brazilian rivers Amazon and Araguaia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evolução do reparo por excisão de nucleotídeo

The phylogeny is one of the main activities of the modern taxonomists and a way to reconstruct the history of the life through comparative analysis of these sequences stored in their genomes aimed find any justification for the origin or evolution of them. Among the sequences with a high level of conservation are the genes of repair because it is important for the conservation and maintenance of genetic stability. Hence, variations in repair genes, as the genes of the nucleotide excision repair (NER), may indicate a possible gene transfer between species. This study aimed to examine the evolutionary history of the components of the NER. For this, sequences of UVRA, UVRB, UVRC and XPB were obtained from GenBank by Blast-p, considering 10-15 as cutoff to create a database. Phylogenetic studies were done using algorithms in PAUP programs, BAYES and PHYLIP package. Phylogenetic trees were build with protein sequences and with sequences of 16S ribosomal RNA for comparative analysis by the methods of parsimony, likelihood and Bayesian. The XPB tree shows that archaeal´s XPB helicases are similar to eukaryotic helicases. According to this data, we infer that the eukaryote nucleotide excision repair system had appeared in Archaea. At UVRA, UVRB and UVRC trees was found a monophyletic group formed by three species of epsilonproteobacterias class, three species of mollicutes class and archaeabacterias of Methanobacteria and Methanococci classes. This information is supported by a tree obtained with the proteins, UVRA, UVRB and UVRC concatenated. Thus, although there are arguments in the literature defending the horizontal transfer of the system uvrABC of bacteria to archaeabacterias, the analysis made in this study suggests that occurred a vertical transfer, from archaeabacteria, of both the NER genes: uvrABC and XPs. According the parsimony, this is the best way because of the occurrence of monophyletic groups, the time of divergence of classes and number of archaeabacterias species with uvrABC system

Estudos ecogenômicos e bioprospectivos de Shewanella ssp

Bacteria trom Shewanella and Geobacter ganera are the most studied iron-reducing microorganisms particularly due to their electron transport systems and contribution to some industrial and environmental problems, including steel corrosion, bioenergy and bioremediation of petroleum-impacted sites. The present study was focused in two ways: the first is an in silico comparative ecogenomic study of Shewanella spp. with sequenced genomes, and the second is an experimental metagenomic work to detect iron-reducing Shewanella through PCR-DGGE of a metabolic gene. The in silico study resulted in positive correIation between copy number of 16S rDNA and genome size in Shewanella spp., with clusters of rrn near lhe origin of replication. This way, the genus is inferred as opportunist. There are no compact genomes and their sequences length varied, ranging from 4306142 nt in S. amazonensis SB2B to 5935403 nt in S. woodyi ATCC 51908, without correIation to temperature range characteristic of each specie. Intragenomic 16S rDNA sequences possess little divergence, but reasonable to resuIt in different phyIogenetic trees, depending on the sequence that is chosen to compare. For moIecuIar detection of iron-reducing Shewanella, it is proposed the mtrB gene as new biomarker. because it codes to a fundamental protein at Fe (III)-reduction. The specific primers were designed and evaluated in silico and resulted in a fragment of 360 pb. In the second study, these primers were tested in a genomic sample from S. oneidensis MR-1, amplifying the expected region. After this successfuI resuIt, the primer set was used as a tool to assess the iron-reducing communities of ShewaneIla genus under an environmental stress, i.e. crude oil contamination in mangrove sediment in Rio Grande do Norte State (Brazil). The primers presented high specificity and the reactions performed resulted in one single band of ampIification in the metagenomic samples. The fingerprinting obtained at DGGE reveaIed temporal variation of Shewanella spp. in analyzed samples. The resuIts presented show the detection of a biotechnological important group of microorganisms, the iron-reducing Shewanella spp. using a metabolic gane as target. It is concluded there are eight or more 16S rDNA sequences in Shewanella genus, with little divergence among them that affects the phylogeny; the pair of primers designed to ampIify mtrB sequences is a viable alternative to detect iron-reducing ShewanelIa in metagenomic approaches; such bacteria are present in the mangrove sediment anaIyzed, with temporal variations in the samples. This is the first experimental study that screened the iron-reducing Shewanella genus in a metagenomic experiment of mangrove sediments subjected to oil contamination through a key metabolic gene

Evolutionary Changes in the Complexity of the Tectum of Nontetrapods: A Cladistic Approach

Background: The tectum is a structure localized in the roof of the midbrain in vertebrates, and is taken to be highly conserved in evolution. The present article assessed three hypotheses concerning the evolution of lamination and citoarchitecture of the tectum of nontetrapod animals: 1) There is a significant degree of phylogenetic inertia in both traits studied (number of cellular layers and number of cell classes in tectum); 2) Both traits are positively correlated accross evolution after correction for phylogeny; and 3) Different developmental pathways should generate different patterns of lamination and cytoarchitecture.Methodology/Principal Findings: The hypotheses were tested using analytical-computational tools for phylogenetic hypothesis testing. Both traits presented a considerably large phylogenetic signal and were positively associated. However, no difference was found between two clades classified as per the general developmental pathways of their brains.Conclusions/Significance: The evidence amassed points to more variation in the tectum than would be expected by phylogeny in three species from the taxa analysed; this variation is not better explained by differences in the main course of development, as would be predicted by the developmental clade hypothesis. Those findings shed new light on the evolution of an functionally important structure in nontetrapods, the most basal radiations of vertebrates.

Caracterização citomorfológica, cultural, molecular e patogênica de Rhizoctonia solani Kühn associado ao arroz em Tocantins, Brasil

No Estado do Tocantins, no Norte do Brasil, a incidência de rizoctoniose no arroz é importante, causando danos significativos em lavouras de arroz irrigado. O principal objetivo deste trabalho foi determinar o grupo de anastomose (AG) de isolados de R. solani associados ao arroz naquela região, testando a hipótese de que esses isolados pertencem ao grupo padrão de anastomose AG-1 IA, que também é o agente causal da mela em soja em áreas úmidas do Norte do Brasil. Todos os quatro isolados de arroz foram caracterizados, através de fusão de hifas, como AG-1 IA. A caracterização cultural, em função das temperaturas basais (mínimas, máximas e ótimas), evidenciou que os isolados de R. solani de arroz apresentaram perfis semelhantes aos padrões AG-1 IA, AG-1 IB e AG-1 IC. Os isolados de arroz foram caracterizados como autotróficos para tiamina assim como os isolados padrões AG-1 IA, IB, IC, AG-4 HGI e o isolado da mela da soja. O teste de patogenicidade em plantas de arroz cultivar IRGA-409 e de patogenicidade cruzada à cultivar IAC-18 de soja (suscetível à mela), indicou que além de causar a queima da bainha em arroz, esses isolados causam mela em soja. da mesma forma, o isolado SJ-047 foi patogênico ao arroz. As seqüências de bases de DNA da região ITS-5.8S do rDNA dos isolados do arroz foram similares às seqüências do AG-1 IA, depositadas no GenBank® - NCBI. A filogenia do ITS-rDNA indicou um grupo filogenético comum formado pelos isolados do arroz, o isolado da soja e o isolado teste do AG-1 IA. Assim, com base em características citomorfológicas, culturais, filogenéticas e patogênicas, foi confirmada a hipótese de que os isolados de R. solani patógenos de arroz do Estado do Tocantins pertencem ao grupo de anastomose AG-1 IA, além da indicação de que esses isolados podem também causar a mela em soja.

Caracterização genética e evolução dos vírus dengue no Estado do Rio Grande do Norte

Dengue is considered as the most important arthropod-borne viral disease throughout the world due to the high number of people at risk to be infected, mainly in tropical and subtropical regions of the planet. The etiologic agent is Dengue Virus (DENV), it is a single positive-stranded RNA virus of the family Flavivirus, genus Flaviviridae. Four serotypes are known, DENV-1, DENV-2, DENV-3 and DENV-4. One of the most important characteristic of these viruses is the genetic variability, which demands phylogenetic and evolutionary studies to understand key aspects like: epidemiology, virulence, migration patterns and antigenic characteristics. The objective of this study is the genetic characterization of dengue viruses circulating in the state of Rio Grande does Norte from January 2010 to December 2012. The complete E gene (1485 pb) of DENV1, 2 e 4 from Brazilian (Rio Grande do Norte) patients was sequenced. Phylogenetic analysis was performed using MEGA 5.2 software, Tamura-Nei model and Neighbor-Joining trees were inferred for the datasets. In Brazil, there is just one DENV-1 genotype (genotype V), one DENV-2 genotype (Asian/American) and two DENV-4 genotypes (genotypes I and II). Brazilian strains of DENV-1 are subdivided in two different lineages (BR-I and BR-II), the Brazilian strains of DENV-2 are subdivided in four lineages (BRI-IV) and genotype II of DENV-4 is subdivided in three Brazilian lineages (BRI-III). The viruses isolated in RN belong to lineage BR-II (DENV-1), BR-IV (DENV-2) and BR-III (DENV-4).The Caribbean and near Latin American countries are the main source of these viruses to Brazil. Amino acids substitutions were detected in three domains of E protein, this makes clear the necessity of studies that associate epidemiological and molecular data to better understand the effects of these mutations. This is the first study about genetic characterization and evolution of Dengue viruses in Rio Grande do Norte, Brazil

Canine distemper virus in a crab-eating fox (Cerdocyon thous) in Brazil: case report and phylogenetic analyses

Although canine distemper is enzootic worldwide and has a wide host range, there are no reports of canine distemper virus there are no reports of canine distemper virus provide information on virus phylogeny and histopathologic lesions. The objective of this study is report and describe canine distemper in a crab-eating fox (C. thous), with a focus oil the phylogeny of the virus strain and the histopathologic lesions in the animal.

The evolution of jumping performance in anurans: morphological correlates and ecological implications

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Caracterização filogenética de amostras do vírus da imunodeficiência felina (FIV) do Estado de São Paulo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Paracoccidioides brasiliensis: phylogenetic and ecological aspects

The knowledge on the biological aspects of Paracoccidioides brasiliensis has evolved greatly since the first description of the disease in 1908. From the pioneers, who were able to clearly demonstrate the fungal nature of the agent, to the recent genomic era, important advances have been achieved. P. brasiliensis is a true fungus, belonging to the Ascomycetous Division, although its sexual phase has not been demonstrated morphologically. A better understanding of the fundamental aspects of the agent, especially its phylogeny and evolutionary history, will provide us with valuable insights allowing a better comprehension of the disease and our capacity to deal with the problem. Concerning the fungus's ecology, although some progress had been observed, the ecological niche of the pathogen has not been determined yet. The aim of the present review is to focus on the biological aspects of P. brasiliensis from an evolutionary point of view, addressing the fungus's phylogenetic aspects, in those special points that might be relevant for the pathogen/host interactions, the biological forces that have been acting on its origin and maintenance of virulence, as well as in determining the fungus's ecology and epidemiology.

Phylogenetic analysis of PRP8 intein in Paracoccidioides brasiliensis species

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)