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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
11

Efeito da atmosfera enriquecida com CO2 em mudas de Psidium guajava L. cv. Pedro Sato / Effect of elevated CO2 on seedlings of Psidium guajava L. cv. Pedro Sato.

Fernanda Mendes Rezende 03 December 2013 (has links)
Psidium guajava é uma espécie tropical, frutífera, de grande importância na fruticultura brasileira e é usada em diversas partes do mundo para o tratamento de doenças que acometem a humanidade. Além de sua importância alimentícia e medicinal, P. guajava também se mostrou boa bioindicadora de poluentes atmosféricos como ozônio e SO2 , pois apresenta respostas características e dose-dependentes das concentrações desses poluentes. Mostrou-se, dessa forma, uma espécie interessante para o estudo de respostas de espécies tropicais a elevadas concentrações de CO2 atmosférico. A fim de avaliar se o elevado CO2 seria uma situação estressante para essa espécie, foi realizada uma fumigação de 130 mudas de P. guajava cv. Pedro Sato em quatro câmaras de topo aberto: duas com ar ambiente ((∼390 ppm) e duas com atmosfera enriquecida com CO2 (∼720 ppm). Foram realizadas coletas quinzenais, retirando-se cinco indivíduos de cada câmara, aleatoriamente, que foram separados em raiz, caule e folhas e imediatamente congelados com nitrogênio líquido. Foram analisados componentes relacionados a sinalização e inativação de espécies reativas de oxigênio em folhas: os teores de poliaminas solúveis, ácido ascórbico e glutationa, além de fenóis, flavonoides, proantocianidinas e taninos totais. Além disso, os extratos metanólicos de folhas foram submetidos à CLAE para análises quali e quantitativas dos principais flavonoides. Parâmetros relacionados ao desenvolvimento da planta também foram analisados, como incremento em altura, massa seca, fração de massa, acúmulo de carbono, açúcares solúveis e amido. As análises de poliaminas, ácido ascórbico, glutationa, e compostos fenólicos de folhas não revelaram alterações significativas entre os tratamentos, exceto para a análise de taninos totais onde foi observado um aumento significativo dessas substâncias nas plantas mantidas a elevado CO2 e após 90 dias de fumigação. A análise por CLAE revelou que a maioria dos flavonoides dessa espécie é derivada de quercetina. Com relação aos parâmetros de crescimento, P. guajava não apresentou diferenças significativas de incremento em altura e massa seca, mas apresentaram acúmulo de amido nas folhas. Analises multivariadas foram utilizadas para integrar as 46 variáveis analisadas. Neste estudo viu-se que a situação de elevado CO2 em mudas de P. guajava é favorável, visto que ocorre acúmulo de amido, substâncias anti-herbivóricas (taninos) e, aparentemente, não há alteração no balanço redox. Contudo, é de extrema importância estudos que avaliem alterações concomitantes de parâmetros ambientais, como aumento de temperatura, disponibilidade de água, ozônio e elevado CO2 . A goiabeira apresenta grande importância econômica relacionada à produção e qualidade de seus frutos, dessa forma são necessários estudos que avaliem o efeito do aumento do CO2 na quantidade e qualidade de frutos produzidos por esta espécie. Por ser uma espécie de crescimento lento, períodos de exposição mais longos são aconselháveis para uma melhor analise da susceptibilidade dessa espécie ao elevado CO2 / Psidium guajava is a tropical fruit species of great importance in Brazilian economy; it is also used in several parts of the world as medicinal. Besides its importance as medicinal and nutritional species, P. guajava has also showed to be a good bioindicator of air pollutants, such as ozone and SO2 . It presents characteristic responses, dose-dependent of the concentrations of these pollutants. Therefore, this species seems to be interesting to study the responses of a tropical species to high concentrations of atmospheric CO2 . In order to assess whether the high CO2 would be a stressful situation for this species, we performed a fumigation experiment using 130 seedlings of P. guajava cv. Pedro Sato in four open-top chambers: two with ambient air (∼ 390 ppm CO2 ) and two with elevated CO2 (∼ 720 ppm). Biweekly, five individuals of each chamber were randomly collected, separated into root, stem and leaves and immediately frozen with liquid nitrogen. Were analyzed parameters related to signaling and inactivation of reactive oxygen species in leaves: levels of soluble polyamines, ascorbate and glutathione, as well as contents of phenols, flavonoids, tannins and proanthocyanidins. Furthermore, the methanol extracts were subjected to HPLC for quantitative and qualitative analysis of the major flavonoids. Parameters related to the development of the plant were also analyzed, as height increment, dry mass, mass fraction, carbon accumulation, soluble sugars and starch accumulation. The analysis of polyamines, ascorbate, glutathione and phenolic compounds of leaves, showed no significant changes when comparing treatments, except for the analysis of tannins which was observed a significant increase of these substances in plants at high CO2 after 90 days of fumigation. HPLC analysis revealed that most of flavonoid composition is quercetin derivatives. The growth parameters showed no significant differences in height increment and dry mass, but the leaves showed accumulation of starch after CO2 fumigation. Multivariate analyzes were used to integrate the 46 variables analyzed. In this study it was seen that the elevated CO2 seem to be favorable to seedlings of P. guajava, once there is an accumulation of starch, antiherbivore substances (tannins) and, apparently, there was no change in the redox balance. However, it is extremely important to perform studies that evaluate concomitant changes in environmental conditions, such as increased temperature, availability of water, ozone and elevated CO2 . Guava has great economic importance in Brazil, fact related to the production and quality of its fruits, so more studies are needed to assess the effect of increased CO2 in the quantity and quality of fruits produced by this species. As a slow-growing species, longer exposure periods are recommended to better analyze the susceptibility of this species to elevated CO2
12

Cell-type specificity and herbivore-induced responses of primary and terpene secondary metabolism in Arabidopsis roots

Zhang, Jingyu 02 September 2013 (has links)
Plants employ diverse defense mechanisms to combat attack by harmful organisms. For instance, plants produce constitutive physical barriers or use chemical compounds such as specialized secondary metabolites to resist herbivore or pathogen invasion. Considering the cost-efficiency and energy balance between defense, growth and reproduction, defense reactions in plants have to be regulated temporally and spatially. As more cost-efficient strategies, plants may induce their defense response only in the presence of the attacker or restrict constitutive defenses to specific tissues or cells. In this study, we investigated aspects of the spatial regulation and induced changes of primary and secondary metabolism in roots of Arabidopsis thaliana. Roots represent important organs for anchoring plants in the soil and taking up water and nutrients. Hence, it is assumed that roots are as well protected as aerial tissues by different defense mechanisms. The first part of this work is focused on the cell-type-specific biosynthesis of volatile terpenes in Arabidopsis roots. Terpenes are the most abundant specialized metabolites in plants and play an important role in plant defense against pathogens or herbivores. Terpene biosynthetic enzyme activities are often coordinated in specific tissues and cellular compartments. Fine-scale transcriptome maps of Arabidopsis roots have shown that terpene biosynthesis is restricted to particular cell types. However, the reasons and significance of this cell-type specificity are not well understood. We hypothesized that the formation of terpene metabolites is not restricted to specific cells but can be supported by different cell types. We, therefore, probed the plasticity of the cell-specific formation of terpenes by swapping the expression of the terpene synthase (TPS) genes, TPS08, TPS13 and TPS25, between different root cell types in the respective mutant background. To investigate the ectopic expression of TPSs at different levels, quantitative real-time PCR (qRT-PCR), confocal microscopy, and gas chromatography-mass spectrometry (GC-MS) were performed. We found that terpene synthase TPS08, which produces the diterpene rhizathalene and is normally expressed in the root vascular tissue, is functionally active when expressed in the epidermis or cortex, although at substantially lower levels compared to the wild type. We did not find an obvious correlation between the volatile emission level and gene transcript level of TPS08, which may be attributed to a reduced activity of the expressed TPS08-yellow fluorescent protein (YFP) fusion protein. When expression of TPS13 (producing the sesquiterpene (Z)-"-bisabolene) was directed from the cortex to the epidermis or stele, TPS13 gene expression and (Z)-"-bisabolene formation was supported by these cell types although to varying levels in comparison to wild type. TPS13-YFP fluorescent signal driven by the epidermal WER and GL3 promoters was primarily detected at the root tip. Terpene production was also observed for the (E)-"-farnesene sesquiterpene synthase TPS25 when its expression was switched from the endodermis and non-hair producing epidermal cells to hair producing epidermal cells although only a weak fluorescent signal was detected from the expressed TPS25-mGFP protein. Together, the results provide preliminary evidence for a relaxed cell specificity of terpene biosynthesis in Arabidopsis roots and suggest that tissue-specific terpene metabolite patterns could change depending on different selective pressures in rhizosphere. In the second part of this study, we performed global gene transcript profiling and primary metabolite analysis of Arabidopsis roots upon feeding by the generalist root herbivore, Bradysia (fungus gnat). In a microarray analysis, we identified 451 of 22,810 genes that were up-regulated more than 2-fold. Gene ontology (GO) analysis showed that 26% of those genes with predicted or known functions play a role in primary or secondary metabolism, while 24% are involved in cell signaling or in responses to stimulating factors, such as jasmonic acid (JA), ethylene, wounding, and oxidative stress. At the metabolite level, we observed only marginal changes of amino acid, sugar and carboxylic acid relative levels over a time course of 4 days of Bradysia feeding. There was a trend for increased levels of amino acids and the relative levels of sucrose were increased significantly ("=0.05) at the fourth day of feeding. In conclusion, the study provided evidence for the induction of genes related to primary and secondary metabolism and stress responses in Arabidopsis roots, but showed only marginal changes at the primary metabolite level. In addition, the work indicated that the formation of terpene-specialized metabolites in Arabidopsis roots is not restricted to specific cells, but can be supported by different cell types. / Master of Science
13

Geny sekundárního metabolismu v půdních bakteriálních společenstvech / Gene pool of the secondary metabolism in soil bacterial communities

Patrmanová, Tereza January 2019 (has links)
The need for new antibiotics and other biologically active compounds is the reason for an increased interest in secondary metabolites of soil bacteria. The phylum Actinobacteria has the dominant position in the soil environment thanks to the potential of producing a broad spectrum of antibiotics and the presence of a number of defense mechanisms preventing the effects of antibiotics. The aim of this thesis was to determine the number of copies of selected secondary metabolic genes in the soils of two sites using designed primers and primers from literature. The design of effective new primers for the detection of selected genes in the soil environment was not achieved in this work, and therefore only primers from literature that had been verified for their specificity were used. In samples taken from soil profiles of two sites, abundances of bacteria, actinobacteria, type II polyketide synthase genes and Erm methyltransferase genes mediating resistance to MLSB antibiotics (macrolides, lincosamides and streptogramins B) were determined by digital PCR. The comparison of the determined copy numbers gave an information about the structure of the bacterial community and the relative abundance of bacteria carrying selected secondary metabolic genes depending on the soil condition changes due to the...
14

Using Small Molecules to Alter Secondary Metabolism in Streptomyces

Ahmed, Salman 10 1900 (has links)
<p>Secondary metabolites produced by bacterial species serve many clinically useful purposes such as anti-bacterial, anti-cancer, and immunosuppresive agents. Actinobacteria, particularly the genus <em>Streptomyces</em>, have been an abundant source of such metabolites for the past half century. The production of secondary metabolites is controlled through vast regulatory cascades, but the activation and control of these pathways is still poorly understood. This leads to the inability to isolate all of the secondary metabolites that <em>Streptomyces</em> are capable of producing. This study focuses on the comparison of synthetic small molecules, which were found to alter the production of secondary metabolites in <em>S. coelicolor</em>. A comparative analysis of two of these molecules, ARC2 and ARC6, shows they modulate secondary metabolites in different ways. In a separate study, ARC2 was shown to achieve this phenotype through the inhibition of a target in fatty acid biosynthesis. The results of this study suggest that ARC6 does not have the same target, although it may target the same metabolic system. Furthermore, these two molecules also have opposite effects on <em>S. coelicolor </em>development. The cumulative results of this study suggest that ARC2 and ARC6 can act as separate chemical tools in enhancing the understanding of secondary metabolism.</p> / Master of Science (MSc)
15

SMALL MOLECULE INTERROGATION OF S. COELICOLOR GROWTH, DEVELOPMENT AND SECONDARY METABOLISM

Craney, Arryn 10 1900 (has links)
<p>Secondary metabolites are vital to human health and strategies to improve their production and detection are equally essential. The blue pigmented metabolite actinorhodin produced by <em>Streptomyces coelicolor</em>, a genus renowned for their diverse secondary metabolites, provides a unique opportunity to identify small molecules probes of secondary metabolism. Small molecules capable of altering secondary metabolism will have widespread application in the streptomycetes due to their ease of addition to any culture condition. Taking advantage of the phenotypic versatility of the <em>S. coelicolor</em> lifecycle, we extended our search for small molecule modulators further to include the entire developmental process. In addition to alterations in secondary metabolism, these processes include growth inhibition, precocious sporulation and alterations in aerial hyphae formation and sporulation. This work provides the foundation for studying <em>Streptomyces</em> by chemical manipulation. Those compounds which stimulate secondary metabolism were narrowed down to 19 ARCs (for antibiotic remodeling compounds). From these, a set of 4 structurally related molecules, the ARC2 series, was identified as weak inhibitors of fatty acid biosynthesis and most likely lead to alterations in secondary metabolism through shifting precursors from primary to secondary metabolism. Consistent with the conservation of fatty acid biosynthesis within bacteria, the effect of the ARC2 series extends in general to the actinomycetes. This provides a simple strategy to alter the secondary metabolic profiles of a diverse range of actinomycetes.</p> / Doctor of Philosophy (PhD)
16

Molecular and Functional Characterization of Terpene Chemical Defense in Arabidopsis Roots in Interaction with the Herbivore Bradysia spp. (fungus gnat)

Vaughan, Martha Marie 18 June 2010 (has links)
Roots and leaves are integrated structural elements that together sustain plant growth and development. Insect herbivores pose a constant threat to both above- and belowground plant tissues. To ward off herbivorous insects, plants have developed different strategies such as direct and indirect chemical defense mechanisms. Research has primarily focused on visible aboveground interactions between plants and herbivores. Root-feeding insects, although often overlooked, play a major role in inducing physical and physiological changes in plants. However, little is known about how plants deploy chemical defense against root herbivores. We have developed an Arabidopsis aeroponic culture system based on clay granulate, which provides access to root tissue and accommodates subterranean insect herbivores. Using this system, feeding performance and plant tissue damage by the root herbivore Bradysia (fungus gnat) were evaluated. Larval feeding was found to reduce Arabidopsis root biomass and water uptake. Furthermore, we have characterized a root-specific terpene synthase AtTPS08, which is responsible for the constitutive formation of the novel volatile diterpene compound, rhizathalene, in Arabidopsis roots. Rhizathalene synthase is a class I diterpene synthase that has high affinity for the substrate geranylgeranyl diphosphate (GGPP) and is targeted to the root leucoplast. Expression of the β-glucuronidase (GUS) reporter gene fused to the upstream genomic region of AtTPS08 demonstrated constitutive promoter activity in the root vascular tissue and root tips. Using the established bioassay with Arabidopsis and Bradysia larvae, in aeroponic culture we could show that roots deficient in rhizathalene synthesis were more susceptible to herbivory. Our work provides in vivo-evidence that diterpene compounds are involved in belowground direct defense against root-feeding insects. Future work is still required to improve our understanding of plant root defense. This study has provided a basis for future investigations on the biochemistry, molecular regulation and defensive function of Arabidopsis root chemicals in interaction with both above- and belowground herbivores (and pathogens). / Ph. D.
17

Leaf Volatile Emissions Structure Tree Community Assembly and Mediate Climate Feedbacks in Tropical Forests

Taylor, Tyeen Colligan, Taylor, Tyeen Colligan January 2017 (has links)
The biochemistry of leaves merges the fates of trees and the atmosphere. Leaf primary metabolism cycles carbon and indirectly drives atmospheric circulation via the latent heat of transpiration. Tropical forests contain half of global forest carbon, and actively cycle carbon and energy year round, making them critical components of the coupled biosphere-climate system. Climate change threatens tropical forests with rising temperatures and increasing variability of precipitation. Their response will influence future biodiversity as well as the fate of the climate. Understanding the physiological attributes that define tropical tree responses and feedbacks to climate is a current research priority. The emission of isoprene gas from plant leaves has been demonstrated to enhance leaf tolerance to high temperatures and drought. Isoprene is a volatile secondary metabolite produced in the chloroplast by approximately one-third of plant species. While the benefits of isoprene are supported by extensive laboratory and greenhouse-based research, work has only begun to explore how the trait is integrated in plant functional strategies. Whether isoprene influences differential species performance and survival across environments has yet to be tested. An impediment to filling this clear ecological research gap has been a lack of instrumentation capable of quantifying isoprene emissions from leaves in remote field settings. The first study presented here tests the hypothesis that isoprene emission influences plant community assembly shifts across environmental gradients and through time in tropical forests. The capacity for a species to produce isoprene was associated with increased relative abundance at higher temperatures and following drought anomalies. A negative relationship with the length of seasonal drought suggests a trade-off between isoprene emission and other plant traits, such as deciduous leaf habit. The second study presents the development of a new instrument that is uniquely optimized for field-based ecological research on leaf volatiles. The new system, named PORCO (Photoionization of Organic Compounds), utilizes custom leaf cuvettes, precision light control, and an optimized commercial photoionization detector to achieve real-time detection of leaf emissions with detection limits better than 0.5 nmol m⁻² leaf s⁻¹. The third study utilizes PORCO to test hypotheses about the structuring of isoprene within plant functional strategies and across forest microenvironments in an eastern Amazonian evergreen tropical forest. The results support the role of isoprene—and potentially other volatile isoprenoids—in mitigating effects of intermittent sun exposure in the sub-canopy. Emissions are structured in a complex, multivariate manner that depends on taxonomy, leaf and wood characteristics, tree height, and light environment. The results from this dissertation work demonstrate that isoprene emission from leaves affects plant responses to climate at ecologically relevant scales. Isoprene influences climate not only by its effect on primary leaf functions, but also by directly altering atmospheric chemistry, and contributing to aerosol and cloud properties. Understanding isoprene's role in forest responses to increasing temperatures and drought will help to predict the feedbacks between forest ecosystems and climatic change.
18

Avaliação da variação do metabolismo secundário da esponja marinha Aplysina fulva em função de sua distribuição geográfica / Evaluation of the secondary metabolism variability within the Aplysina fulva marine sponge related to its geographic distribution

Pereira, Fábio Renato 26 October 2006 (has links)
Estudos realizados em 1979 por Kelecom e Kanengiesser mostraram que extratos brutos obtidos a partir de amostras da esponja marinha Aplysina fulva não possuíam derivados bromados. Estes foram resultados inesperados, uma vez que esponjas pertencentes à ordem Verongida são conhecidos produtores de metabólitos derivados da dibromotirosina. O presente projeto teve como meta a investigação química de extratos obtidos de duas amostras de A. fulva, sendo uma coletada em São Sebastião (SP) e a outra em Angra dos Reis (RJ), objetivando verificar a ocorrência de derivados da bromotirosina e uma possível variabilidade química dependendo da distribuição geográfica das esponjas. Sete derivados da dibromotirosina foram isolados das duas amostras de Aplysina fulva: quatro compostos da amostra de Angra dos Reis e três da de São Sebastião. As estruturas dos compostos foram identificadas por análises espectroscópicas (como IV, UV, RMN 1H, RMN 13C, HMQC, COSY, HMBC) e de espectrometria de massas, e ainda por comparação com dados da literatura. Os resultados obtidos confirmaram que os derivados da dibromotirosina são bons marcadores quimiotaxonômicos para as esponjas da Ordem Verongida. Além disso, a variabilidade química observada para a A. fulva parece ser influência de fatores abióticos e bióticos como sazonalidade, disponibilidade de nutrientes, ou associação com diferentes micro-organismos. / Studies developed in 1979 by Kelecom and Kanengiesser showed that crude extracts obtained from samples of the marine sponge Aplysina fulva were devoid of bromotyrosine derivatives. These results were rather unexpected, since sponges belonging to the Order Verongida are well-known producers of bromotyrosine-derived metabolites. The present project aimed the chemical investigation of crude extracts obtainded from two samples of A. fulva, one collected at São Sebastião (SP) and the second from Angra dos Reis (RJ), in order to verify the occurrence of bromotyrosine derivatives and a possible chemical variability depending on the geographical distribution of the sponge. Seven bromoyrosine derivatives have been isolated from the two samples of A. fulva: four compounds from the Angra dos Reis sample and three from the São Sebastião sample. The structures of the compounds have been established by spectroscopic analysis, (including IV, UV, RMN 1H, RMN 13C, HMQC, COSY, HMBC) and mass spectrometry, as well as by comparison with literature data. The results obtained confirmed that bromotyrosine derivatives are good chemotaxonomic markers for sponges of the Order Verongida. Moreover, it appears that chemical variability of A. fulva may be influenced by abiotic and biotic factors, such as seasonality, nutrients availability, or association with distinct micro-organisms.
19

Bioluminescência fúngica: papel ecológico, purificação e clonagem de enzimas / Fungal bioluminescence: ecological role, purification and cloning of enzymes

Waldenmaier, Hans Eugene 21 December 2016 (has links)
Esta tese de doutorado descreve os estudos realizados para elucidar a biologia molecular da bioluminescência fúngica e sua relevância ecológica na natureza. A recente descoberta de que a luciferina fúngica é a 3-hidroxihispidina permitiu a caracterização do metabolismo secundário da fenilalanina nos genomas recém-sequenciados e transcriptomas de micélios das espécies luminescentes Panellus stipticus e Neonothopanus gardneri. Adicionalmente os genomas e transcriptomas de variedades não luminescente de P. stipticus e Lentinula edodes serviram como respectivos controles. Em geral, os genes envolvidos no metabolismo secundário da fenilalanina em amostras luminescentes tinham expressão igual ou superior àquela de espécies não luminescentes. Um agrupamento de genes relacionados com a biossíntese de fenilalanina foi encontrado em ambos os genomas luminescentes e não luminescentes de P. stipticus. A abundância de genes transcritos neste agrupamento foi semelhante para as espécies luminescentes e não luminescentes de P. stipticus, mas a policetídeo sintase tipo I em P. stipticus não luminescentes foi significativamente sub-regulada. Não foi encontrado agrupamento semelhante nos genomas de N. gardneri e L. edodes, sendo que os correspondentes homólogos estavam espalhados em diferentes loci. Extratos de fungos podem ser preparados in vitro, com a adição de 3-hidroxihispidina para produzir luz verde em abundância. A preparação de extratos proteicos de luciferase foi melhorada e a estrutura da luciferase, parcialmente purificada, foi investigada por espectrometria de massas. A presença de luciferase nos géis de purificação foi revelada usando-se luciferina e molécula similares à luciferina advindas de extratos de plantas. O nicho ecológico nas vizinhas de cogumelos bioluminescentes foi investigado de duas maneiras, armadilhas adesivas com cogumelos artificiais de acrílico, iluminados com luz LED verde e através da observação direta de cogumelos bioluminescentes com fotografia no infravermelho com lapso de tempo. Os estudos ecológicos foram conduzidos nos biomas da Mata Atlântica e da Mata dos Cocais, no Brasil. Baratas, aranhas, tesourinhas, grilo e vagalumes tec-tecs foram os animais mais comuns que interagiram com os cogumelos. Todos estes animais podem agir como dispersores de propágulos e, em alguns casos, como defensores dos cogumelos. / This PhD thesis describes the studies performed to elucidate the molecular biology of fungal bioluminescence and the ecological significance of the trait in the wild. The recent discovery that the fungal luciferin is 3-hydroxyhispidin has allowed for the characterization of phenylalanine secondary metabolism in the newly sequenced genomes and mycelium transcriptomes of luminescent Panellus stipticus and Neonothopanus gardneri, additionally the genomes and transcriptomes of a non-luminescent variety of P. stipticus and Lentinula edodes served as respective controls. In general the genes involved in phenylalanine secondary metabolism had greater or equal expression in luminescent samples than non luminescent. A cluster of genes related to the secondary metabolism of phenylalanine was found in both luminescent and non luminescent P. stipticus genomes. Transcript abundance of genes in this cluster was similar in both luminescent and non-luminescent Panellus stipticus, but the type I polyketide synthase in non luminescent Panellus stipticus was significantly down regulated. A similar gene cluster in the N. gardneri and L. edodes genomes was absent with corresponding homologues scattered at different genomic loci. Cell free fungal extracts can be combined in vitro with the addition of 3-hydroxyhispidin to produce abundant green light. Preparation of proteinaceous luciferase extracts was improved and partially purified luciferase samples were investigated by mass spectrometry. The presence of luciferase in the separation gel was also evidenced by using luciferin and luciferin-like molecules from plant extracts. The ecological niche surrounding bioluminescent mushrooms was investigated through two main means, glue traps with acrylic mushroom facsimiles that were internally illuminated with green LED lights and direct observation of bioluminescent mushrooms with infrared time lapse photography. Ecological studies were performed in the Atlantic rainforest (Mata Atlântica) and transitional Coconut Palm forest (Mata dos Cocais) biomes of Brazil. Cockroaches, spiders, earwigs, crickets, and luminescent click beetles were the most common animal interacting with mushrooms. All of these animals may be acting as fungal propagule dispersers and in some cases defense of the mushroom.
20

Genômica comparativa de Microcystis aeruginosa (Cyanobacteria: Chroococcales), com ênfase em genes envolvidos com síntese de produtos naturais / Comparative genomics of Microcystis aeruginosa (Cyanobacteria: Chroococales), with emphasis on genes related to natural product synthesis

Weiss, Bruno 04 April 2017 (has links)
A ampla diversidade metabólica das cianobactérias é associada não somente a sua importância nos ciclos biogeoquímicos, mas também a sua distribuição global. Tal característica também é responsável pela capacidade destes organismos em produzir uma ampla variedade de substâncias de estruturas incomuns e atividades de interesse para o homem. Microcystis é um gênero cianobacteriano reconhecido como produtor de mais de duas centenas de produtos naturais, incluindo cianotoxinas. Microcystis aeruginosa é uma espécie frequentemente encontrada em florações, portanto causando preocupações sobre sua influência ecológica, especialmente em corpos d\'água doce utilizados para consumo humano. Desta forma, o objetivo deste trabalho foi o levantamento da diversidade e quantidade de metabólitos secundários que podem ser produzidos pela espécie, através de análises genômicas, além de variáveis que podem potencialmente interferir nas análises computacionais, procurando-se por padrões na espécie, e comparando-se 18 linhagens de todos os continentes. Foi encontrado o total de 235 agrupamentos relacionados ao metabolismo secundário, categorizados em 12 classes segundo as estruturas de seus produtos, nas 18 linhagens, evidenciando a riqueza de agrupamentos relacionados ao metabolismo secundário encontrados nesta espécie. Destes agrupamentos, os mais abundantes pertencem às categorias dos Terpenos, Híbridos, Bacteriocinas e NRPS. Entre as NRPS, nenhuma foi comum a todas as linhagens. Ainda, a quantidade de agrupamentos variou entre 6 e 21, e a quantidade de categorias de produtos variou entre 4 e 10, mostrando uma distribuição heterogênea de agrupamentos e tipos de metabólitos preditos. Esta distribuição heterogênea foi detalhada para melhor compreensão deste padrão encontrado na espécie. Dos agrupamentos de NRPS, os três mais frequentes foram selecionados para uma análise pormenorizada de sua estrutura e sequência: aeruginosina (15 linhagens), microcistina (11 linhagens), e micropeptina (15 linhagens). O agrupamento de micropeptina encontrado nas linhagens SPC777, TAIHU98 e PCC 9806 se mostrou amplamente dissimilar com relação à referência utilizada, potencialmente indicando um erro de identificação causado pela plataforma antiSMASH utilizada para a localização dos agrupamentos. Análises de colinearidade genômica mostram uma baixíssima sintenia entre os genomas das linhagens em análise, sugerindo frequentes eventos de reorganização genômica. Ainda, análises de pangenoma mostram um cenário em que mais genomas desta espécie são necessários para a estimativa da quantidade total de genes diferentes que a espécie pode possuir, o que é interessante para futuros estudos de procura de metabólitos secundários. Análises do genoma cerne apontam para uma estimativa segura de 1.944 genes comuns a todos os genomas desta espécie, o que corresponde entre 35% e 50% dos genes em cada linhagem. Análises estatísticas apontam para diferentes graus de interferência não linear da quantidade de sequências contíguas na observação de diferentes padrões de outras características genômicas, sugerindo precaução nas expectativas com relação ao metabolismo secundário em caso de linhagens em que a montagem gênica ultrapasse o limite superior aproximado de 100 sequências contíguas. / The wide metabolic diversity of cyanobacteria is associated not only with their importance in biogeochemical cycles, but also with their global distribution. Such a feature is also responsible for the ability of these organisms to produce a wide variety of substances with unusual structures and activities of interest to man. Microcystis is a cyanobacterial genus recognized as a producer of more than two hundred natural products, including cyanotoxins. Microcystis aeruginosa is a species frequently found in cyanobacterial blooms, thus causing concerns about its ecological influence, especially in freshwater bodies used for human consumption. In this way, the objective of this work was the survey of the diversity and quantity of secondary metabolites that can be produced by the species, through genomic analyzes, besides variables that can potentially interfere in the computational analyzes, searching for patterns in the species, and comparing 18 strains from all the continents. A total of 235 clusters, categorized in 12 classes according to the structure of their products, were found in the 18 strains, evidencing the richness of clusters related to the secondary metabolism found in this species. Of these clusters, the most abundant belong to the categories of Terpenes, Hybrids, Bacteriocins and NRPS. Among NRPS, none were common to all strains. Also, the number of groups ranged from 6 to 21, and the number of product categories ranged from 4 to 10, showing a heterogeneous distribution of predicted groupings and types of metabolites. Such a heterogeneous distribution was detailed for a better understanding of this pattern found in the species. Of the NRPS clusters, the three most frequent were selected for a detailed analysis of their structure and sequence: aeruginosin (15 strains), microcystin (11 strains), and micropeptin (15 strains). The micropeptide cluster found in the SPC777, TAIHU98 and PCC 9806 strains was widely dissimilar to the reference, só potentially indicating an identification error caused by the antiSMASH platform used to locate the clusters. Genomic collinearity analyzes showed a very low synteny among the genomes of the strains under analysis, suggesting frequent events of genomic reorganization. Also, pangenome analyzes show a scenario in which more genomes of this species are needed for the estimation of the total amount of different genes the species may possess, which is interesting for future studies conserning secondary metabolites. Coregenome analyzes point to a reliable estimate of 1,944 genes common to all genomes of this species, which corresponds to 30% up to 50% of the genes in each strain. Statistical analyzes point to different degrees of non-linear interference of the number of contiguous sequences on the observation of different patterns of other genomic characteristics, suggesting necessary caution about expectations regarding the secondary metabolism in case of strains in which the gene assembly exceeds the approximate upper limit of 100 contiguous sequences.

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