Análise filogenética e funcional de dois genes de reparo homólogos a AP endonuclease em cana-de-açúcar: ScARP1 e ScARP3

The genome of all organisms constantly suffers the influence of mutagenic factors from endogenous and/or exogenous origin, which may result in damage for the genome. In order to keep the genome integrity there are different DNA repair pathway to detect and correct these lesions. In relation to the plants as being sessile organisms, they are exposed to this damage frequently. The Base Excision DNA Repair (BER) is responsible to detect and repair oxidative lesions. Previous work in sugarcane identified two sequences that were homologous to Arabidopsis thaliana: ScARP1 ScARP3. These two sequences were homologous to AP endonuclease from BER pathway. Then, the aim of this work was to characterize these two sequence using different approaches: phylogenetic analysis, in silico protein organelle localization and by Nicotiana tabacum transgenic plants with overexpression cassette. The in silico data obtained showed a duplication of this sequence in sugarcane and Poaceae probably by a WGD event. Furthermore, in silico analysis showed a new localization in nuclei for ScARP1 protein. The data obtained with transgenic plants showed a change in development and morphology. Transgenic plants had slow development when compared to plants not transformed. Then, these results allowed us to understand better the potential role of this sequence in sugarcane and in plants in general. More work is important to be done in order to confirm the protein localization and protein characterization for ScARP1 and ScARP3

Análise proteômica de Chromobacterium violaceum: acúmulo estacionário e diferencial após exposição à luz UVC

Chromobacterium violaceum is a free-living bacillus, Gram-negative commonly found in water and sand of tropical and subtropical regions. One of its main characteristic it's the ability to produce the purple pigment named violacein, that shows countless biological activities. In 2003, the genome of this organism was totally sequenced and revealed important informations about the physiology of this bacteria. However, few post-genomics studies had been accomplished. This work evaluated the protein profile of C. violaceum cultivated in LB medium at 28ºC that allowed the identification and characterization of proteins related to a possible secretion system that wasn't identified and characterized yet in C. violaceum, to the quorum sensing system, to regulatory process of transcription and translation, stress adaptation and biotechnological potential. Moreover, the response of the bacteria to UVC radiation was evaluated. The comparison of the protein profile, analyzed through 2-D electrophoresis, of the control group versus the treatment group allowed the identification of 52 proteins that arose after stress induction. The obtained results enable the elaboration of a stress response pathway in C. violaceum generated by the UVC light. This pathway, that seems to be a general stress response, involves the expression of proteins related to cellular division, purine and pirimidine metabolism, heat chock or chaperones, energy supply, regulation of biofilm formation, transport, regulation of lytic cycle of bacteriophages, besides proteins that show undefined function. Despite the response present similarities with the classic SOS response of E. coli, we still cannot assert that C. violaceum shows a SOS-like response, mainly due to the absence of characterization of a LexA-like protein in this organism

Proteômica diferencial da Chromobacterium violaceum em resposta ao estresse oxidativo induzido por peróxido de hidrogênio

The β-proteobacterium Chromobacterium violaceum is a Gram-negative, free-living, saprophytic and opportunistic pathogen that inhabits tropical and subtropical ecosystems among them, in soil and water of the Amazon. It has great biotechnological potential, and because of this potential, its genome was completely sequenced in 2003. Genome analysis showed that this bacterium has several genes with functions related to the ability to survive under different kinds of environmental stresses. In order to understand the physiological response of C. violaceum under oxidative stress, we applied the tool of shotgun proteomics. Thus, colonies of C. violaceum ATCC 12472 were grown in the presence and absence of 8 mM H2O2 for two hours, total proteins were extracted from bacteria, subjected to SDS-PAGE, stained and hydrolysed. The tryptic peptides generated were subjected to a linear-liquid chromatography (LC) followed by mass spectrometer (LTQ-XL-Orbitrap) to obtain quantitative and qualitative data. A shotgun proteomics allows to compare directly in complex samples, differential expression of proteins and found that in C. Violaceum, 131 proteins are expressed exclusively in the control condition, 177 proteins began to be expressed under oxidative stress and 1175 proteins have expression in both conditions. The results showed that, under the condition of oxidative stress, this bacterium changes its metabolism by increasing the expression of proteins capable of combating oxidative stress and decreasing the expression of proteins related processes bacterial growth and catabolism (transcription, translation, carbon metabolism and fatty acids). A tool with of proteomics as an approach of integrative biology provided an overview of the metabolic pathways involved in the response of C. violaceum to oxidative stress, as well as significantly amplified understanding physiological response to environmental stress. Biochemical and "in silico" assays with the hypothetical ORF CV_0868 found that this is part of an operon. Phylogenetic analysis of superoxide dismutase, protein belonging to the operon also showed that the gene is duplicated in genome of C. violaceum and the second copy was acquired through a horizontal transfer event. Possibly, not only the SOD gene but also all genes comprising this operon were obtained in the same manner. It was concluded that C. violaceum has complex, efficient and versatile mechanisms in oxidative stress response

A rede de marcação e recaptura: o caso de borboletas do gênero heliconius

(The Mark and Recapture Network: a Heliconius case study). The current pace of habitat destruction, especially in tropical landscapes, has increased the need for understanding minimum patch requirements and patch distance as tools for conserving species in forest remnants. Mark recapture and tagging studies have been instrumental in providing parameters for functional models. Because of their popularity, ease of manipulation and well known biology, butterflies have become model in studies of spatial structure. Yet, most studies on butterflies movement have focused on temperate species that live in open habitats, in which forest patches are barrier to movement. This study aimed to view and review data from mark-recapture as a network in two species of butterfly (Heliconius erato and Heliconius melpomene). A work of marking and recapture of the species was carried out in an Atlantic forest reserve located about 20km from the city of Natal (RN). Mark recapture studies were conducted in 3 weekly visits during January-February and July-August in 2007 and 2008. Captures were more common in two sections of the dirt road, with minimal collection in the forest trail. The spatial spread of captures was similar in the two species. Yet, distances between recaptures seem to be greater for Heliconius erato than for Heliconius melpomene. In addition, the erato network is more disconnected, suggesting that this specie has shorter traveling patches. Moving on to the network, both species have similar number of links (N) and unweighed vertices (L). However, melpomene has a weighed network 50% more connections than erato. These network metrics suggest that erato has more compartmentalized network and restricted movement than melpomene. Thus, erato has a larger number of disconnected components, nC, in the network, and a smaller network diameter. The frequency distribution of network connectivity for both species was better explained by a Power-law than by a random, Poissom distribution, showing that the Power-law provides a better fit than the Poisson for both species. Moreover, the Powerlaw erato is much better adjusted than in melpomene, which should be linked to the small movements that erato makes in the network

Filogenia molecular de fungos gasteroides das ordens Phallales e Geastrales (Phallomycetidae)

Gasteroids fungi are characterized by the basidiospores maturation inside the basidioma, from which spores liberation occurs in a passive manner. These fungi were once seen as a well definite class of Basidiomycota, but nowadays they are considered an artificial assemblage, because the organisms have independent evolutionary histories forming a polyphyletic group with a vast morphological variety. Despite their diversity, studies with this group in the tropics are incipient, and the phylogenetic relationships of the species from temperate climate remain unknown. Thus, this work aimed to elucidate the phylogenetic relationships of gasteroids fungi from the Geastrales and Phallales orders, with the inclusion of tropical and temperate species, and with these analyses suggest a systematic position of species like Aseroë floriformis and Phallus roseus, as well as to verify if the lignicolous habit can indicate parental relationship in the Geastrum genus. For this, basidiomata were collected at Atlantic rain forest areas, during the rainy season, and the specimen identification followed specific literature for gasteroid fungi. The phylogenetic analyses were performed with Maximum Parsimony and Bayesian Analysis, making use of RPB2 and 28S nuclear genes and atp6 mitochondrial gene. It could be observed on the Phallales dendogram, that Aseroë floriformis did not cluster with A. rubra, and that it has an anterior divergence from all others species of the family Clathraceae used in this analysis, assuming a basal position in the clade. Phallus roseus, which once was recognized as Itajahya, has previous divergence from the group formed by Phallus species. At the Geastrales dendogram, in the group corresponding to Geastrum genus, it could be observed that species with lignicolous habitat clustered in a clade with high support values. So, the results suggest the creation of a new genus to accommodate A. floriformis, and the revalidation of Itajahya, as well as it can be affirmed that the lignicolous habitat on the Geastrum genus in fact indicates parental relationships, and that it has arised only once at the evolutionary history of the genus

Estrutura da população de Microcerotermes exiguus ( Isoptera: Termitidae) e sua partuicipação no consumo de madeira em uma área de caatinga, Nordeste, Brasil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Avaliação de parâmetros de estresse oxidativo em plantas de cana-de-açúcar tratadas com peróxico de hidrogênio

The genus Saccharum belongs to Poaceae family. Sugarcane has become important monocultures in Brazil due to their products: ethanol and sugar. The production may change between different regions from Brazil. This difference is related to soil, climatic conditions and temperature that promotes oxidative stress that may induce an early flowering. The aim of this work was to identify the effects of oxidative stress. In order to analyse this, sugarcane plants were submitted to oxidative stress using hydrogen peroxide. After this treatment, the oxidative stress were analyzed Then, the plant responses were analyzed under different approaches, using morphophysiological, biochemical and molecular tools. Thus, sugarcane plants were grown under controlled conditions and until two months they were subjected first to a hydroponics condition for 24 hours in order to acclimation. After this period, these plants were submitted to oxidative stresse using 0 mM, 10 mM, 20 mM and 30 mM hydrogen peroxide during 8 hours. The histomorphometric analysis allowed us to verify that both root and leaf tissues had a structural changes as it was observed by the increased in cell volume, lignin accumulation in cell walls. Besides, this observation suggested that there was a change in redox balance. Also, it was analyzed the activity of the SOD, CAT and APX enzymes. It was observed an increase in the SOD activity in roots and it was also observed a lipid peroxidation in leaves and roots. Then, in order to identify proteins that were differently expressed in this conditions it was used the proteomic tool either by bidimensional gel or by direct sequencing using the Q-TOF EZI. The results obtained with this approach identified more than 3.000 proteins with the score ranging from 100-5000 ions. Some of the proteins identified were: light Harvesting; oxygenevolving; Thioredoxin; Ftsh-like protein Pftf precusor; Luminal-binding protein; 2 cys peroxiredoxin e Lipoxygenase. All these proteins are involved in oxidative stress response, photsynthetic pathways, and some were classified hypothetical proteins and/or unknown (30% of total). Thus, our data allows us to propose that this treatment induced an oxidative stress and the plant in response changed its physiological process, it made changes in tissue, changed the redox response in order to survival to this new condition

Produção, acúmulo e decomposição da serapilheira em uma restinga da Ilha do Mel, Paranaguá, PR, Brasil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Isolation and characterization of Flavobacterium columnare (Bernardet et al. 2002) from four tropical fish species in Brazil

Flavobacterium columnare é o agente etiológico da columnariose em peixes de água doce, ocasionando enfermidade na pele e nas brânquias, provocando freqüentemente um grande número de mortalidade. O objetivo deste estudo foi o isolamento e a caracterização de Flavobacterium columnare em peixes tropicais no Brasil. Piracanjuba (Brycon orbignyanus), pacu (Piaractus mesopotamicus), tambaqui (Colossoma macropomum) e cascudo (Hypostomus plecostomus) foram examinados externamente com relação a sinais característicos de columnariose, como manchas acinzentadas na cabeça, região dorsal e pedúnculo caudal dos peixes. A amostragem compreendeu a coleta de 50 exemplares de peixes, representando as quatro diferentes espécies escolhidas para este estudo. Amostras para o isolamento foram obtidas através de raspado com swab estéril das lesões e do rim dos peixes clinicamente diagnosticados como acometidos por columnarios e imediatamente semeados em meios de culturas artificiais (líquido e sólido) próprios para o estudo de Flavobacterium segundo Carlson e Pacha (1968). No meio líquido, houve o desenvolvimento de microrganismos que observados em gota pendente apresentaram a forma de bacilos finos, longos, móveis por deslizamento. Através da coloração de Gram, apresentaram morfologia de bacilos finos, Gram negativos, agrupados em colunas. em meio sólido, as colônias eram pequenas, cinza-amareladas, com borda em forma de raiz. No total, foram obtidos quatro isolamentos: 01 cepa de Brycon orbignyanus; 01 cepa de Piaractus mesopotamicus; 01 cepa de Colossoma macropomum; e 01 cepa de Hypostomus plecostomus. A caracterização bioquímica das amostras, como absorção do vermelho Congo, produção de flexirrubina, produção de H2S e redução do nitrato, sugere que os isolamentos poderiam ser classificados como Flavobacterium columnare.

Ant community richness and composition across a gradient from Eucalyptus plantations to secondary Atlantic Forest

As florestas secundárias e plantações de espécies exóticas estão se expandindo nas paisagens tropicais. No entanto, nossa compreensão sobre o valor destas florestas para a conservação da biodiversidade de invertebrados ainda é incipiente. Neste trabalho, usamos a fauna de formigas de serapilheira para avaliar a diversidade desses insetos entre três florestas de Eucalyptus, sendo uma comercial (quatro anos de idade) e duas abandonadas em diferentes idades de regeneração (16 e 31 anos) e uma área de Mata Atlântica secundária. A riqueza total foi mais alta na floresta secundária e nos plantios de Eucalyptus abandonados há mais tempo. A densidade de espécies na floresta secundária foi significativamente maior quando comparado as plantações de Eucalyptus, mas não difere entre eucaliptais; análise de ordenação revelou diferenças na composição de espécies entre as plantações de Eucalyptus com subbosque ausente e com subbosque desenvolvido ou em desenvolvimento. Ainda, foi constatada uma sobreposição acentuada entre amostras de serapilheira das florestas de eucaliptos abandonadas há mais tempo e a floresta secundária. em geral, plantações de eucalipto foram caracterizadas pela presença de espécies generalistas e de ampla distribuição. Nossos resultados indicam que embora o subbosque de plantações de eucaliptos com maior idade de regeneração suporte um conjunto relativamente alto de espécies generalistas de formigas, é improvável que eucaliptais conservem a maioria das espécies de florestas primárias, especialmente predadores especializados, Dacetini e espécies nômades.

Qualidade de frutos de cajá-manga armazenados sob diferentes temperaturas

O presente trabalho teve como objetivo determinar o efeito de diferentes temperaturas de armazenamento na qualidade de frutos de cajá-manga. Os frutos de cajá-manga foram obtidos junto ao CEAGESP - Companhia de Entrepostos e Armazéns Gerais de São Paulo, e transportados em caixas isotérmicas para o Laboratório onde foram selecionados quanto à aparência, estádio de maturação, ausência de danos físicos, depois sanitizados em solução de 50 ppm de cloro livre e acondicionados em bandejas de poliestireno expandido, envolto por filme de policloreto de vinila (PVC). O delineamento experimental utilizado foi o inteiramente casualizado, com três tratamentos (4ºC, 8ºC e 25ºC) e 5 épocas de avaliação (0; 3; 6; 9 e 12 dias após a instalação do experimento). em cada coleta, foram avaliadas a firmeza, a acidez titulável, os sólidos solúveis, o teor de ácido ascórbico, a cor da casca e, também, a liberação de CO2 pelos frutos ao longo do tempo. Foi observado que baixas temperaturas prolongam a vida útil dos frutos e que a temperatura de armazenamento influencia nas características avaliadas, sendo a temperatura de 8ºC a mais adequada para o armazenamento dos frutos de cajá-manga.

Banco de sementes de plantas daninhas e sua correlação com a flora estabelecida no agroecossistema cana-crua

O conhecimento da composição específica do banco de sementes de plantas daninhas e a sua correlação com a flora estabelecida são importantes para nortear o manejo a ser adotado e a escolha dos herbicidas. A colheita mecanizada de cana-de-açúcar acarretou mudanças significativas na composição da flora infestante, quando proporcionou a manutenção de uma camada de palha, reduziu a movimentação do solo e dispensou a prática da queimada. Foram realizados levantamentos do banco de sementes e da flora de plantas daninhas que se estabeleceu em 28 talhões colhidos mecanicamente, sem queima prévia da palha. Com base nos dados de banco de sementes, efetuaram-se estudos fitossociológicos e de correlação entre a composição do banco de sementes e a flora emergida. As principais espécies presentes no banco de sementes foram as pertencentes à classe das dicotiledôneas anuais, com destaque para Amaranthus spp. e diversas espécies de Euphorbiaceae e Convolvulaceae. As sementes de gramíneas tradicionais da cultura tiveram pouca participação. O banco de sementes apresentou correlação não-significativa com a flora emergente, independentemente da época de colheita do talhão, da metodologia de quantificação do banco de sementes e das espécies de plantas daninhas.

Estimativa da área foliar de plantas daninhas: Brachiaria decumbens Stapf. e Brachiaria brizantha (Hochst.) Stapf

Com o objetivo de obter uma equação que, através de parâmetros lineares dimensionais das folhas, permita a estimativa da área foliar de Brachiaria decumbens Stapf. e Brachiaria brizantha (Hochst.) Stapf., estudaram-se correlações entre a área foliar real (Sf) e parâmetros dimensionais do limbo foliar, como o comprimento ao longo da nervura principal (C) e a largura máxima (L), perpendicular à nervura principal. Todas as equações, exponenciais, geométricas ou lineares simples, permitiram boas estimativas da área foliar. do ponto de vista prático, sugere-se optar pela equação linear simples envolvendo o produto C x L, considerando o coeficiente linear igual a zero. Desse modo, a estimativa da área foliar de B. decumbens pode ser feita pela fórmula Sf = 0,9810 x (C x L), ou seja, 98,10% do produto entre o comprimento ao longo da nervura principal e a largura máxima, enquanto que, para a B. brizantha a estimativa da área foliar pode ser feita pela fórmula SF = 0,7468 x (C x L), ou seja 74,68% do produto entre o comprimento ao longo da nervura principal e a largura máxima da folha.

Caracterização dos marcadores químicos e avaliação de atividades biológicas do extrato de Spondias mombim

Spondias mombin is a fruitful species dispersed in tropical regions of America, Africa and Asia. In Brazil, the species can be found mainly in the northern and northeastern regions. Scarce chemical and pharmacological studies have been reported for S. mombin and until this moment studies about chemical markers were not developed. In this context, the aims of this study were to characterize the chemical markers from S. mombin leaves and evaluate their anti-inflammatory, antioxidant and antiproliferative potentials. The chemical profile of the hydroethanolic extract from S. mombin leaves analyzed by HPLC-DAD, through a validated method, allowed the identification and quantification of ellagic acid and chlorogenic acid. This extract showed anti-inflammatory potential in acute peritonitis model induced by carrageenan. The hydroethanolic extract from S. mombin leaves was subjected to a liquid-liquid partition with the solvents: n-hexane, dichloromethane, ethyl acetate and n-butanol. Regarding the anti-inflammatory potential of the fractions obtained they were active; however, ethyl acetate fraction at 200 mg/kg showed highlighted results. The compounds ellagic acid and chlorogenic acid also inhibited the leukocyte migration to the site of inflammation at 2.5, 5 and 10 mg/kg. The hydroethanolic extract, fractions and the chemical markers showed significant antioxidant potential when evaluated in different assays: DPPH Free-Radical Scavenging, Superoxide Radical Scavenging, Hydroxyl Radicals Scavenging and Reducing Power. Taken together our results showed that hydroethanolic extract of S. mombin leaves has ellagic acid and chlorogenic acid as bioactive markers and it demonstrated antiinflammatory and antioxidant properties besides no cytotoxicity against 3T3 cells. It enables us to suggest S. mombin as an important species to develop herbal drugs

Fitossociologia de comunidades de plantas daninhas em agroecossistema cana-crua

Objetivou-se neste trabalho estudar a fitossociologia de comunidades de plantas daninhas de canaviais colhidos no sistema mecanizado, sem queima prévia da palha, e a similaridade entre talhões quanto à composição de espécies. Os levantamentos foram realizados em 28 talhões comerciais na região de Ribeirão Preto-SP. em cada talhão foram demarcadas áreas de coleta e avaliação, na proporção de duas por hectare, mantidas sem controle, e que serviram de local para as amostragens de plantas daninhas. As amostragens foram feitas com quadrados vazados (0,5 x 0,5 m), lançados aleatoriamente duas vezes em cada uma das áreas. Essas amostragens foram realizadas determinando-se a densidade e a biomassa específica aos 120 dias após o corte da cana. Cyperus rotundus foi a principal espécie, destacando-se quanto aos valores de importância relativa (IR). As plantas dicotiledôneas anuais de propagação por sementes também se destacaram, dentre as quais diversas espécies das famílias Euphorbiaceae e Convolvulaceae. em contrapartida, as gramíneas tradicionais de áreas de cana colhida queimada tiveram pouco destaque. O índice de Shannon (H) de diversidade de espécies das comunidades variou de 0 a 1,61, e o índice de similaridade entre os talhões (S) foi muito variável. A maioria das espécies ou grupo de espécies apresentou padrão agregado (V/m > 1,00), com valores relativamente altos de índice de agregação (V/m). Entretanto, na maioria dos casos, Cyperus rotundus e as Convolvulaceas apresentaram os maiores índices.