Caracterização in situ e diversidade genética de algodoeiros mocós (Gossypium hissutum raça marie galante) da Região Nordeste do Brasil

Brazil is one of the major centers of diversity for polyploid cotton plants; these plants belong to the genus Gossypium, which has three known species: G. hirsutum, G. barbadense and G. mustelinum. The Northeast is the only region where the three species occur, the last group being endemic. Northeast s cotton plants can be important sources of variability for genetic breeding. It is believed that great part of local diversity is being lost, due to economic, political, cultural and agricultural problems. In an attempt to mitigate this loss and delineate conservation strategies it is necessary to know how the species are found where they occur. The objective was to characterize and determine how plants are maintained in situ in the states of Maranhão, Piauí, Ceará, Rio Grande do Norte and Paraíba at the beginning of the XXI century. The in situ characterization of G. hirsutum and G. barbadense was conducted through structured interviews with the cotton plants owners and through the analysis of the environment. The data were collected during expeditions undertaken between the years 2004 to 2005. Twenty-two plants were collected in the state of Paraíba, forty-four in the state of Rio Grande do Norte, one hundred and forty-six in the state of Ceará, forty in the state of Maranhão and ninety-one plants in the state of Piauí. All plants collected in the states of Paraíba and Rio Grande do Norte belonged to moco type. Moco cotton plants also predominated in the other states, representing 92%, 62% and 78% of plants collected in Ceará, Piauí and Maranhão, respectively. The other cotton plants collected belong to the species G. barbadense. The cotton plants were found in situ as dooryard plants, roads side, feral populations, cultivation or local varieties. Great part were dooryard plants (45.2%), being major in Piauí and Maranhão. Cultivation predominated in Ceará; in Rio Grande do Norte feral populations were the most frequent and, in Paraíba, local varieties. The maintenance of moco plants is related, mainly, to the phytotherapic domestic use (20.9%) and to confection of lamp wicks (29.7%). Few inhabitants in Paraíba, Rio Grande do Norte, Piauí and none in Maranhão used harvest the plants, storage the seeds or gin; however, in Ceará, 40.5% of owners affirmed that they harvested and commercialized the fiber. It was found that the maintenance of species is dependent of the fragile cultural habits of local inhabitants, therefore the maintenance in situ is not a suitable way to conservation of genetic resources. The efforts must be directed to the continuity of collections, maintenance and characterization ex situ

caracterização in situ e estrutura genética de populações de Gossypium mustelinum

Gossypium mustelinum Miers ex Watt is the only cotton species native from Brazil. It is endemic of the semi-arid region from North-east of the country, where it occur near from resilient water sources. The threats to the in situ conservation of the populations are caused by human interference in its habitat, mainly by excessive cattle graze and deforestation. Establish efficient strategies of in situ conservation depend on the accomplishment of a diagnosis of how the specie is found in its natural environment, and the knowledge about the genetic structure of the populations. The objectives of this work were i) to determine the in situ conditions of two populations present in rivers from basin of Rio Paraguaçu at the Bahia State, ii) to evaluate the structure and genetic variability presented in both populations, iii) to establish in situ and ex situ conservation strategies. It were realized collection in november 2007, when was realized in situ characterization of G. mustelinum. SSR markers were used for analyze 218 genotypes deriving from two populations of the G. mustelinum, localized at Tocó river and the Capivara river. The allelic frequencies, the heterozigosity and the F statics were estimated. All the plants were classified as wild and natives, and there was no evidence of the use the plants or its parts. The populations showed different conservation conditions in situ. Few plantlets were found in sites with excessive cattle feed, an indication that the damages in young plants should be high enough to compromise the renovation of the populations. On the other hand, populations were well preserved when the anthropic damages was low or inexistent. The 14 SSR primer pairs amplified 17 loci with a medium number of 5 alleles per locus (a total of 85 alleles). The high level of endogamy estimated (FIS=0,808) and the low observed heterozygosity (H0=0,093) were indicatives that the populations reproduce mainly by selfing, geitonogamy and crosses between related individuals. The genetic diversity was high (HE=0,482) and the differentiation between the populations was very high (FST=0,328). At least two sites from both populations of G. mustelinum must be preserved to achieve suitable in situ conservation. Actions that preserve the gallery forest and keep the cattle away should implemented, and could be as simple as erecting a fence. It is not possible anticipated if the in situ preservation will be possible. Therefore collections and ex situ preservation of representative specimens are essential to conserve the genetic diversity of native G. mustelinum

Estrutura genética de populações de Amphobotrys ricini, agente causal do mofo cinzento da mamoneira

The gray mold, causal organism Amphobotrys ricini, is one of the major diseases of castor bean. Difficulties in managing plant disease arises form the limited understanding of the genetic structure of A. ricini, their complexity and variability make it difficult to control. Genetic structure can be used to infer the relative impact of different forces that influence the evolution of pathogen populations, that allow to predict the potencial for pathogen populations to envolve in agricultural ecosystems. Growers protect their crop by applying fungicides, but there aren t fungicides to provide significant control of gray mold of castor bean. The objectives of this work were use RAPD to determine the genetic structure of A. ricini subpopulations in Paraíba and assay the sensitivity of A. ricini isolates to azoxystrobin and carbendazim. To determine the genetic structure of A. ricini subpopulations in Paraíba, 23 isolates were colleted from two different geographic location (subpopulation). These isolates were analysed by RAPD using 22 random decamer primers, purchased from OPERON, produced a total of 80 markers polimorphics. The resulting matrixes were analysed using PopGene version 1.32. Sensitivity to azoxystrobin and carbendazim of 30 isolates, colleted form Paraíba and Alagoas, was estimated based on spore germination and colony growth inhibition. The stock solutions were added toV8 medium after sterilization to produce final concentrations of 0, 0.01, 0.1, 1, 10, and 100 µg/ml of carbendazim and 0, 0.001, 0.01, 0.1, 1, and 10 µg/ml of azoxystrobin. All statistical analyses were performed using SAS to estimate the dose that inhibited fungal growth by 50% (ED50 values). The genetic diversity within subpopulations (Hs=0,271) accounted for 92% of the total genetic diversity (Ht=0,293), while genetic diversity between subpopulations (Gst = 0,075) represented only 7,5%. The estimated number of migrants per generation (NM ) was 6,15. Nei s average gene identity across 80 RAPD loci was 0,9468. Individual ED50 values, for the 30 isolates screened for their sensitivity to azoxystrobin, ranged From a maximum of 0,168 µg/ml to a minimum of 0,0036 µg/ml. The ED50 values for carbendazim varied within the range of 0,026 to 0,316 µg/ml

Genética e biologia reprodutiva de vriesea minarum (Bromeliaceae): em busca de estratégias de conservação no quadrilátero ferrífero, minas gerais

Vriesea minarum is a rupiculous bromeliad species, with naturally fragmented populations, restricted to the Iron Quadrangle, Minas Gerais, Brazil. It is a threatened species, which is suffering from habitat loss due to the growth of cities and mining activities. The knowledge of genetic variability in plant populations is one of the main branches of conservation genetics, linking genetic data to conservation strategies while the knowledge about plant reproductive biology can aid in understanding key aspects of their life story, as well as in the comprehension of their distribution and survival strategies. Thus, the study of diversity, richness, and genetic structure, as well as the reproductive biology of populations of V. minarum can contribute to the development of conservation actions. Chapter 1 presents the transferability of 14 microsatellite loci for V. minarum. Among the results of this chapter, we highlight the successful transferability of 10 microsatellite loci described for other species of Bromeliaceae, all of which are polymorphic. In Chapter 2, we present the genetic analyses of 12 populations of V. minarum that are distributed throughout the Iron Quadrangle. We used the 10 microsatellite loci tested in Chapter 1. The results show a low population structuring (Fst = 0.088), but with different values of genetic richness (mean = 2.566) and gene diversity (mean = 0.635) for all populations; and a high inbreeding coefficient (Gis = 0.376). These may be the result of pollinators action and/or efficient seed dispersal, thus allowing a high connectivity among populations of naturally fragmented outcrops. The reproductive biology and floral morphology of a population of V. minarum, located in the Parque Estadual da Serra do Rola-Moça, are studied in Chapter 3. This reserve is the only public environmental protection area where the species occurs. As a result of field experiments and observations, we found that the species has its flowering period from January to March, with flowers that last for two days and that it has a mixed pollination syndrome. It is primarily alogamous, but also has the capacity to be self-ferilized. It is expected that data obtained in chapters 1, 2 and 3 serve as basis for other studies with species from the ferruginous rocky fields, since until now, to our knowledge, there are no other survey of endemic species from the Iron Quadrangle, seeking to merge the genetic knowledge, with the data of the reproductive biology, with the ultimate aim of biodiversity conservation. Considering the great habitat loss for the species by mining, it becomes crucial to analyze the creation of new protected areas for its conservation

Análise da estrutura genética populacional da piraúna (cephalopholis fulva: serranidae) ao longo da costa e ilhas oceânicas brasileiras

Brazil has about 8,500 km of coastline and on this scale, fishing is a historically important source of animal protein for human consumption. The national fishing background shows a growth of marine fishery production until 1985 and within this period it was recorded a steady decline. From the year 2003 fishing statistics aim to some "recovery" of the total fisheries production, which probably is related to a change in industry practice. The target of commercial fishing became smaller species with low commercial value, but very abundants. The coney, Cephalopholis fulva (Serranidae), is one of these species that have been suffering a greater fishing pressure in recent years. In order to provide data about the current situation of the genetic diversity of these populations, several molecular markers have been being used for this purpose. The prior knowledge of genetic variability is crucial for management and biodiversity conservation. To this end, the control region sequences (dloop) of mtDNA from Cephalopholis fulva (Serranidae) from five geographical points of the coast of Brazil (Ceará, Rio Grande do Norte, Bahia and Espírito Santo) and the Archipelago of Fernando de Noronha (FN) were sequenced and their genetic diversity analyzed. The FST values were very low (0.0246 to 0.000), indicating high gene flow between the sampled spots. The indices h and indicate a secondary contact between previously allopatric lineages differentiated or large and stable populations with long evolutionary history. Tests of Tajima and Fu showed expansion for all populations. In contrast, the mismatch distribution and SSD indicated expansion just for coastal populations. Unlike other species of the Atlantic which have been deeply affected by events on later Pleistocene, the population-genetic patterns of C. fulva may be related to recent events occurred approximately 130,000 years ago. Moreover, the data presented by geographical samples of the specie C. fulva showed high genetic diversity, also indicating the absence of deleterious effects of over-exploitation on this specie, as well as evidence of complete panmixia between all sampled populations

Conservação genética da orquídea Cattleya granulosa Lindley

Cattleya granulosa Lind is a large and endemic orchid in Atlantic Forest fragments in Northeast Brazil. The facility of collecting, uniqueness of their flowers, which have varying colors between green and reddish brown, and distribution in coastal areas of economic interest make their populations a constant target of predation, which also suffer from environmental degradation. Due to the impact on their populations, the species is threatened. In this study, we evaluate the levels of spatial aggregation in a preserved population, analyze the phylogenetic relationships of C. granulosa Lindl. with four other Laeliinae species (Brassavola tuberculata, C. bicolor, C. labiata and C. schofieldiana) and also to evaluate the genetic diversity of 12 remaining populations of C. granulosa Lindl. through ISSR. There was specificity of epiphytic C. granula Lindl. with a single host tree, species of Eugenia sp. C. granulosa Lindl. own spatial pattern, with the highest density of neighbors within up to 5 m. Regarding the phylogenetic relationships and genetic patterns with other species of the genus, C. bicolor exhibited the greatest genetic diversity (HE = 0.219), while C. labiata exhibited the lowest level (HE = 0.132). The percentage of genetic variation among species (AMOVA) was 23.26%. The principal component analysis (PCA) of ISSR data showed that unifoliate and bifoliolate species are genetically divergent. PCA indicated a close relationship between C. granulosa Lindl. and C. schofieldiana, a species considered to be a variety of C. granulosa Lindl. by many researchers. Population genetic analysis using ISSR showed all polymorphic loci. The high genetic differentiation between populations (ФST = 0.391, P < 0.0001) determined the structure into nine groups according to log-likelihood of Bayesian analysis, with a similar pattern in the dendrogram (UPGMA) and PCA. A positive and significant correlation between geographic and genetic distances between populations was identified (r = 0.794, P = 0.017), indicating isolation by distance. Patterns of allelic diversity suggest the occurrence of population bottlenecks in most populations of C. granulosa Lindl. (n = 8). Genetic data indicate that enable the maintenance of genetic diversity of the species is complex and is directly related to the conservation of different units or groups that are spatially distant.

Desenvolvimento de marcadores microssatélites e estrutura genética espacial da Copernicia prunifera (Miller) H. E. Moore (Arecaceae)

The present study aimed to develop microsatellite markers (SSR) for Copernicia prunifera; and characterize the demographic pattern and the spatial genetic structure (SGS) in different development stages of C. prunifera in a natural population of Rio Grande do Norte (RN) by using ISSR molecular markers. 17 SSR primers pairs were developed, which were tested by using DNA from samples of different populations. The demographic and genetic spatial structure was assessed in a plot with an area of 0.55 ha, where all individuals were georeferenced. The molecular analyses with the use of microsatellite markers pointed out that all built primers pairs, when submitted to PCR, had amplification. They showed sizes of base pairs ranging between 113 and 250 bp. The demographic analyses showed a clustered standard of spatial distribution in the first distance classes, random between 40 and 50 m and segregated in higher distances. Eight ISSR primers were used, thereby producing a total of 102 loci, with 100 of them being polymorphic. Among the three stages, the young showed the highest Nei’s genetic diversity index (He = 0.37); whilst the lowest index was found in the reproductive adults (He = 0.34). The AMOVA results showed a greater genetic differentiation within the development stages (98.61%) in comparison to the interval among the stages (1.39%). The total population (n = 161) showed a positive and significant relationship of kinship in the first distance class (12.3 m). The young showed a significant kinship up to 10.5 m and negative in the fifth distance class (37.6 m). The non-reproductive adults had a positive relationship of kinship in the first distance class (11.0 m) and random distribution of genotypes in the remaining classes. The reproductive adults showed genotypes spatially distributed in a random way. The values for the genetic bottleneck tests proved that the number of loci with excess observed heterozygosity was greater than expected. The SGS results reflect the restricted dispersion of the species, and the bottleneck tests reflect the reduction genotypes provoked by the anthropization of natural environments of C. prunifera.

Variabilidade genética de Leishmania spp. circulantes entre seres humanos e cães e infecção de flebotomíneos em áreas endêmicas para as leishmanioses no Rio Grande do Norte

Leishmania infantum is the main etiologic agent of visceral leishmaniasis in the New World. The pattern of distribution of leishmaniasis has changed substantially and has presented an emerging profile within the periphery of the Large Urban Centers. Leishmania infection can compromise skin, mucosa and viscera. Only 10% of the individuals infected develop the disease and 90% of human infection is asymptomatic. The main factors involved in the development of the disease are the host immune response, the vector’s species and the parasite’s genetic content. The sequencing of Leishmania isolated seeks to increase the understanding of the symptoms of individuals. The aim of this study was to evaluate the genetic diversity of circulating Leishmania strains among humans, and symptomatic and asymptomatic, and dogs from endemic areas of Rio Grande do Norte State and analyze sandflies from endemic areas for cutaneous and visceral disease. The genetic variability was evaluated by the use of markers hsp70 , ITS1 and a whole genome sequencing was also carried out. The amplified hsp70 and ITS1 of samples were analyzed and assembled using a Phred / Phrap package. The dendograms were constructed using the same methodology, but adding 500 bootstraps, followed by inferences on the relationships between Leishmania variants. The sequences of the 20 Brazilian isolates were mapped to the reference genome L. infantum JPCM5, using the Bowtie2 program and the identification of 36 contigs. The information of the valid SNPs were used in the PCA. SNPs were visualized by Geneious 7.1 and IGV. The genome annotations were transferred to their respective chromosomes and displayed on Geneious. The matching sequences of all chromosomes were aligned using Mauve. The phylogenetic trees were calculated according to maximum likelihood and JTT models. Sandflies were analyzed by PCR for the identification of Leishmania infection, a blood meal source and GAPDH sand fly. As a result, hsp70 and ITS1 were not capable of identifying genetic variability among human isolates from symptomatic and asymptomatic, and dogs. The complete sequencing of the 20 Brazilian isolates revealed a strong similarity between the circulating Leishmania strains in Rio Grande do Norte. The isolates collected in the city of Natal from humans and canines remained grouped in all analyzes, suggesting that there is genotypic and geographic proximity among the isolates. The isolated samples in the 1990s had a higher genotypic diversity when compared to freshly isolated samples. All isolates presented 36 chromosomes with variable ploidy among them, no correlation was found between the number of amastina genes copies, gp63, A2 and SSG with such clinic forms. In general, we did not find correlation between symptomatic and asymptomatic clinical forms and the gene content of the Brazilian isolates of Leishmania. 34,28% of the sandflies collected in the upper west region were L. longipalpis and the main sources of blood meal were humans, dogs and chickens.

Variabilidade genética de Leishmania spp. circulantes entre seres humanos e cães e infecção de flebotomíneos em áreas endêmicas para as leishmanioses no Rio Grande do Norte

Leishmania infantum is the main etiologic agent of visceral leishmaniasis in the New World. The pattern of distribution of leishmaniasis has changed substantially and has presented an emerging profile within the periphery of the Large Urban Centers. Leishmania infection can compromise skin, mucosa and viscera. Only 10% of the individuals infected develop the disease and 90% of human infection is asymptomatic. The main factors involved in the development of the disease are the host immune response, the vector’s species and the parasite’s genetic content. The sequencing of Leishmania isolated seeks to increase the understanding of the symptoms of individuals. The aim of this study was to evaluate the genetic diversity of circulating Leishmania strains among humans, and symptomatic and asymptomatic, and dogs from endemic areas of Rio Grande do Norte State and analyze sandflies from endemic areas for cutaneous and visceral disease. The genetic variability was evaluated by the use of markers hsp70 , ITS1 and a whole genome sequencing was also carried out. The amplified hsp70 and ITS1 of samples were analyzed and assembled using a Phred / Phrap package. The dendograms were constructed using the same methodology, but adding 500 bootstraps, followed by inferences on the relationships between Leishmania variants. The sequences of the 20 Brazilian isolates were mapped to the reference genome L. infantum JPCM5, using the Bowtie2 program and the identification of 36 contigs. The information of the valid SNPs were used in the PCA. SNPs were visualized by Geneious 7.1 and IGV. The genome annotations were transferred to their respective chromosomes and displayed on Geneious. The matching sequences of all chromosomes were aligned using Mauve. The phylogenetic trees were calculated according to maximum likelihood and JTT models. Sandflies were analyzed by PCR for the identification of Leishmania infection, a blood meal source and GAPDH sand fly. As a result, hsp70 and ITS1 were not capable of identifying genetic variability among human isolates from symptomatic and asymptomatic, and dogs. The complete sequencing of the 20 Brazilian isolates revealed a strong similarity between the circulating Leishmania strains in Rio Grande do Norte. The isolates collected in the city of Natal from humans and canines remained grouped in all analyzes, suggesting that there is genotypic and geographic proximity among the isolates. The isolated samples in the 1990s had a higher genotypic diversity when compared to freshly isolated samples. All isolates presented 36 chromosomes with variable ploidy among them, no correlation was found between the number of amastina genes copies, gp63, A2 and SSG with such clinic forms. In general, we did not find correlation between symptomatic and asymptomatic clinical forms and the gene content of the Brazilian isolates of Leishmania. 34,28% of the sandflies collected in the upper west region were L. longipalpis and the main sources of blood meal were humans, dogs and chickens.

Análise metagenômica da microbiota de ambientes aquáticos do estado do Rio Grande do Norte - Brasil

The screening for genes in metagenomic libraries from soil creates opportunities to explore the enormous genetic and metabolic diversity of microorganisms. Rivers are ecosystems with high biological diversity, but few were examined using the metagenomic approach. With this objective, a metagenomic library was constructed from DNA soil samples collected at three different points along the Jundiaí-river (Rio Grande do Norte-Brazil). The points sampled are from open area, rough terrain and with the direct incidence of sunlight. This library was analyzed functionally and based in sequence. For functional analysis Luria-Bertani solid medium (LB) with NaCl concentration varied from 0.17M to 0.85M was used for functional analysis. Positives clones resistant to hypersaline medium were obtained. The recombinant DNAs were extracted and transformed into Escherichia coli strain DH10B and survival curves were obtained for quantification of abiotic stress resistance. The sequences of clones were obtained and submitted to the BLASTX tool. Some clones were found to hypothetical proteins of microorganisms from both Archaea and Bacteria division. One of the clones showed a complete ORF with high similarity to glucose-6-phosphate isomerase which participates in the synthesis of glycerol pathway and serves as a compatible solute to balance the osmotic pressure inside and outside of cells. Subsequently, in order to identify genes encoding osmolytes or enzymes related halotolerance, environmental DNA samples from the river soil, from the water column of the estuary and ocean were collected and pyrosequenced. Sequences of osmolytes and enzymes of different microorganisms were obtained from the UniProt and used as RefSeqs for homology identification (TBLASTN) in metagenomic databases. The sequences were submitted to HMMER for the functional domains identification. Some enzymes were identified: alpha-trehalose-phosphate synthase, L-ectoina synthase (EctC), transaminase L-2 ,4-diaminobutyric acid (EctB), L-2 ,4-diaminobutyric acetyltransferase (EctA), L-threonine 3 dehydrogenase (sorbitol pathway), glycerol-3-phosphate dehydrogenase, inositol 3-phosphate dehydrogenase, chaperones, L-proline, glycine betaine binding ABC transporter, myo-inositol-1-phosphate synthase protein of proline simportadora / PutP sodium-and trehalose-6-phosphate phosphatase These proteins are commonly related to saline environments, however the identification of them in river environment is justified by the high salt concentration in the soil during prolonged dry seasons this river. Regarding the richness of the microbiota the river substrate has an abundance of halobacteria similar to the sea and more than the estuary. These data confirm the existence of a specialized response against salt stress by microorganisms in the environment of the Jundiaí river

Metagenômica: busca de novos genes envolvidos com a biodegradação de hidrocarbonetos

Industrial activities, oil spills and its derivatives, as well as the incomplete combustion of fossil fuels have caused a great accumulation of hydrocarbons in the environment. The number of microorganisms on the planet is estimated at 1030 and prokaryotes the most abundant. They colonized diverse environments for thousands of years, including those considered extreme and represent an untapped source of metabolic and genetic diversity with a large biotechnological potential. It is also known that certain microorganisms have the enzymatic capacity to degrade petroleum hydrocarbons and, in many ecosystems, there is an indigenous community capable of performing this function. The metagenomic has revolutionized the microbiology allowing access uncultured microbial communities, being a powerful tool for elucidation of their ecological functions and metabolic profiles, as well as for identification of new biomolecules. Thus, this study applied metagenomic approaches not only for functional selection of genes involved in biodegradation and emulsification processes of the petroleum-derived hydrocarbons, but also to describe the taxonomic and metabolic composition of two metagenomes from aquatic microbiome. We analyzed 123.116 (365 ± 118 bp) and 127.563 sequences (352 ± 120 bp) of marine and estuarine metagenomes, respectively. Eight clones were found, four involved in the petroleum biodegradation and four were able to emulsify kerosene indicating their abilities in biosurfactants synthesis. Therefore, the metagenomic analyses performed were efficient not only in the search of bioproducts of biotechnological interest and in the analysis of the functional and taxonomic profile of the metagenomes studied as well

Uso sustentável da Copernicia prunifera (Miller) H. E Moore no semiárido potiguar: valorização de saberes e conservação dos recursos genéticos

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Expressão heteróloga de biossurfactantes identificados em bibliotecas metagenômicas

The microorganisms have a vast genetic diversity and they are present throughout the biosphere, however, only about 1% of the species can be cultivated by traditional cultivation techniques. Within this diversity there is a huge pool genetic and biological being explored. The metagenomics has enabled direct access to microbial genome derived from environmental samples using independent methods of cultivation. The methodology enables to obtain functional information about the proteins, as well as identify potential products with biotechnological interest and new industrially exploitable biological resources, such as new solutions to environmental impacts. Oil-contaminated areas are characterized by a large accumulation of hydrocarbons and surfactants may be used for bioremediation. Thus, the metagenomic approach was used in this study in order to select genes involved in the degradation and hydrocarbon emulsification. In a previous work, the environmental DNA (eDNA) was extracted from soil samples collected from two different areas (Caatinga and Saline River) of Rio Grande do Norte (Brazil), the metagenomic libraries were constructed and functionally analyzed. The clone able to degrade the oil was evaluated for the ability to synthesize biosurfactants. The sequence analysis revealed an ORF with 897 bp, 298 amino acids and a protein with around 34 kDa. The search for homology in GenBank revealed sequence similarity with a hypothetical protein of representatives Halobacteriaceae family, who were recently shown as strains producing biosurfactants. The presence of the inserted coding sequence and the acquired phenotype was confirmed. Primers were designed and the ORF amplified by PCR. The ORF was subcloned into pETDuet-1 expression vector for subsequent purification of the protein of interest containing a histidine tail. The tests performed to confirm the biosurfactant activity and the ability of hydrocarbon degradation showed positive results. The immunodetection test (western blot) using the monoclonal AntiHis® confirmed the presence of the environmental protein. This study was the first to report a possible protein with biosurfactant activity obtained from a metagenomic approach

Otimização do controle do diagrama de radiação de radares de varredura para rastreio de foguetes usando o método GAMMC para o Caso Planar (GAMMC-P)

Launching centers are designed for scientific and commercial activities with aerospace vehicles. Rockets Tracking Systems (RTS) are part of the infrastructure of these centers and they are responsible for collecting and processing the data trajectory of vehicles. Generally, Parabolic Reflector Radars (PRRs) are used in RTS. However, it is possible to use radars with antenna arrays, or Phased Arrays (PAs), so called Phased Arrays Radars (PARs). Thus, the excitation signal of each radiating element of the array can be adjusted to perform electronic control of the radiation pattern in order to improve functionality and maintenance of the system. Therefore, in the implementation and reuse projects of PARs, modeling is subject to various combinations of excitation signals, producing a complex optimization problem due to the large number of available solutions. In this case, it is possible to use offline optimization methods, such as Genetic Algorithms (GAs), to calculate the problem solutions, which are stored for online applications. Hence, the Genetic Algorithm with Maximum-Minimum Crossover (GAMMC) optimization method was used to develop the GAMMC-P algorithm that optimizes the modeling step of radiation pattern control from planar PAs. Compared with a conventional crossover GA, the GAMMC has a different approach from the conventional one, because it performs the crossover of the fittest individuals with the least fit individuals in order to enhance the genetic diversity. Thus, the GAMMC prevents premature convergence, increases population fitness and reduces the processing time. Therefore, the GAMMC-P uses a reconfigurable algorithm with multiple objectives, different coding and genetic operator MMC. The test results show that GAMMC-P reached the proposed requirements for different operating conditions of a planar RAV.

Análise metagenômica da microbiota de ambientes aquáticos do estado do Rio Grande do Norte - Brasil

The screening for genes in metagenomic libraries from soil creates opportunities to explore the enormous genetic and metabolic diversity of microorganisms. Rivers are ecosystems with high biological diversity, but few were examined using the metagenomic approach. With this objective, a metagenomic library was constructed from DNA soil samples collected at three different points along the Jundiaí-river (Rio Grande do Norte-Brazil). The points sampled are from open area, rough terrain and with the direct incidence of sunlight. This library was analyzed functionally and based in sequence. For functional analysis Luria-Bertani solid medium (LB) with NaCl concentration varied from 0.17M to 0.85M was used for functional analysis. Positives clones resistant to hypersaline medium were obtained. The recombinant DNAs were extracted and transformed into Escherichia coli strain DH10B and survival curves were obtained for quantification of abiotic stress resistance. The sequences of clones were obtained and submitted to the BLASTX tool. Some clones were found to hypothetical proteins of microorganisms from both Archaea and Bacteria division. One of the clones showed a complete ORF with high similarity to glucose-6-phosphate isomerase which participates in the synthesis of glycerol pathway and serves as a compatible solute to balance the osmotic pressure inside and outside of cells. Subsequently, in order to identify genes encoding osmolytes or enzymes related halotolerance, environmental DNA samples from the river soil, from the water column of the estuary and ocean were collected and pyrosequenced. Sequences of osmolytes and enzymes of different microorganisms were obtained from the UniProt and used as RefSeqs for homology identification (TBLASTN) in metagenomic databases. The sequences were submitted to HMMER for the functional domains identification. Some enzymes were identified: alpha-trehalose-phosphate synthase, L-ectoina synthase (EctC), transaminase L-2 ,4-diaminobutyric acid (EctB), L-2 ,4-diaminobutyric acetyltransferase (EctA), L-threonine 3 dehydrogenase (sorbitol pathway), glycerol-3-phosphate dehydrogenase, inositol 3-phosphate dehydrogenase, chaperones, L-proline, glycine betaine binding ABC transporter, myo-inositol-1-phosphate synthase protein of proline simportadora / PutP sodium-and trehalose-6-phosphate phosphatase These proteins are commonly related to saline environments, however the identification of them in river environment is justified by the high salt concentration in the soil during prolonged dry seasons this river. Regarding the richness of the microbiota the river substrate has an abundance of halobacteria similar to the sea and more than the estuary. These data confirm the existence of a specialized response against salt stress by microorganisms in the environment of the Jundiaí river