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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.
771

The effects of defoliation on seasonal growth dynamics, the importance of internal nitrogen-recycling and the availability of soil nutrients: implications for the invasive potential of Buddleia davidii (Franch.)

Thomas, Marc Merlin January 2007 (has links)
ABSTRACT Assessing the impact of herbivory on plant growth and reproduction is important to predict the success of biocontrol of invasive plants. Leaf area production is most important, as photosynthesis provides the foundation for all plant growth and fitness. High levels of defoliation generally reduce the productivity of plants. However, leaf area production fluctuates during the season and compensational growth may occur, which both complicate accurate estimations of defoliation impacts. Under field conditions the interaction with neighbouring species and the availability of soil nutrients need to be assessed in order to gauge long term effects of weed invasions on natural environments. In this thesis I have investigated seasonal leaf area dynamics in Buddleia davidii following repeated artificial defoliation, to quantify compensational leaf production and to understand the regulatory mechanisms involved. The impact of defoliation on photosynthesis, seed production, germination and nitrogen translocation patterns were analysed. Finally, possible facilitation between B. davidii and a native nitrogen fixer, Coriaria arborea, and the impact of B. davidii on soil nutrient availability were investigated. In defoliated B. davidii, increased node production (34%), leaf size (35%) and leaf longevity (12%) resulted in 52% greater total emergent leaf area in the short term. However, with time and diminishing tissue resources the compensation declined. No upregulation of photosynthesis was observed in pre-existing leaves. Compensational leaf area production occurred at the expense of reproduction but the germination capacity of individual seeds was unaffected. In B. davidii, nitrogen reserves are stored in old leaves. Thus, the defoliation-induced decline in tissue reserves led to changes in the remobilisation pattern and increased the importance of soil uptake but biomass production especially that of roots had declined significantly (39%). Slight facilitation effects from the neighbouring nitrogen fixer and VA-mycorrhizae were observed on B. davidii in the field, while its impact on soil chemistry during spring was negligible. Defoliation of B. davidii resulted in priority allocation of resources to compensational leaf growth and a concomitant reduction in flower and seed production. The compensational leaf production greatly increased the demand for nitrogen, while continued leaf removal decreased the pool of stored nitrogen. This led to changes in nitrogen remobilisation and an increased importance of root uptake. However, the significant decline in root growth will likely impair adequate nutrient uptake from the soil, which is especially important where B. davidii invades nutrient poor habitats and will increase the success of biocontrol of the species. While mycorrhizae increase nutrient accessibility for B. davidii, it is likely that the additional stress of defoliation will negate the small facilitative effects from nitrogen-fixing species like C. arborea. This research provides new insights into the mechanisms regulating leaf area dynamics at the shoot level and systemic physiological responses to defoliation in plants, such as nitrogen translocation. The compensation in leaf area production was considerable but only transitory and thus, the opportunity to alleviate effects of leaf loss though adjustment of light capture limited. However, to ascertain that photosynthesis at whole plant level does not increase after defoliation, more detailed measurements especially on new grown leaves are necessary. In general, defoliation had greatly reduced plant growth and performance so that an optimistic outlook for controlling this species can be given. Conclusions about the wider impacts of B. davidii on soil chemistry and community function will require further research.
772

Mimicking the Outer Coordination Sphere in [FeFe]-Hydrogenase Active Site Models : From Extended Ligand Design to Metal-Organic Frameworks

Pullen, Sonja January 2017 (has links)
Biomimetic catalysis is an important research field, as a better understanding of nature´s powerful toolbox for the conversion of molecules can lead to technological progress. [FeFe]-hydrogenases are very efficient catalysts for hydrogen production. These enzymes play a crucial role in the metabolism of green algae and certain cyanobacteria. Their active site consists of a diiron complex that is embedded in an interactive protein matrix. In this thesis, two pathways for mimicking the outer coordination sphere effects resulting from the protein matrix are explored. The first is the construction of model complexes containing phosphine ligands that are coordinated to the iron center as well as covalently linked to the bridging ligand of the complex. The effect of such linkers is an increased energy barrier for the rotation of the Fe(CO2)(PL3)-subunit, which potentially could stabilize a terminal hydride that is an important intermediate in the proton reduction cycle. The second pathway follows the incorporation of [FeFe]-hydrogenase active site model complexes into metal-organic frameworks (MOFs). Resulting MOF-catalysts exhibit increased photocatalytic activity compared to homogenous references due to a stabilizing effect on catalytic intermediates by the surrounding framework. Catalyst accessibility within the MOF and the influence of the framework on chemical reactivity are examined in the work presented. Furthermore, an initial step towards application of MOF-catalysts in a device was made by interfacing them with electrodes. The work of this thesis highlights strategies for the improvement of biomimetic model catalysts and the knowledge gained can be transferred to other systems mimicking the function of enzymes.
773

Corticular Photosynthetic Dynamics for a Coastal Evergreen Shrub: Myrica Cerifera

Vick, Jaclyn K. 01 January 2007 (has links)
I quantified seasonal variations in corticular photosynthesis in 1st through 5th order branches of Myrica cerifera L. (Myricaceae) in order to determine whether corticular photosynthesis contributes to whole plant carbon gain by reducing respirational CO2 loss. Maximum % refixation was 110 ± 39 % of CO2 efflux in the dark (Rd) in 1st order branches during winter, minimum was 18 ± 3 % in 5th order branches during summer. Variations in % refixation paralleled changes in photosynthetically active radiation (PAR). As light attenuated with increasing branch order % refixation decreased. Increased PAR in the winter due to a more sparse canopy lead to increases in % refixation. Total chlorophyll content and chlorophyll a:b ratios were consistent with shade acclimation as branch order increased. Corticular photosynthesis may be a mechanism to enhance shrub expansion due to increased whole plant carbon use efficiency (CUE) and water use efficiency (WUE) attributed to refixation.
774

Analýza antioxidačních systémů, morfologických a fyziologických parametrů u dvou odrůd bobu (Vicia faba L.) během stresu suchem / Analysis of antioxidant systems, morphological and physiological parameters in two faba bean (Vicia faba L.) cultivars during drought stress

Jelénková, Iva January 2013 (has links)
Knowledge of physiological and morfological mechanisms which are related to drought tolerance is essential in breeding high-yielding and tolerant plants, which would not suffer unfavourable environmental conditions. Drought stress can lead to oxidative damage which causes serious disorders in physiological and biochemical processes in plant cells. Antioxidants can reduce those negative effects. Morphological (dry mass of shoot and roots, height of shoot, number of leaves), physiological (photosynthetic efficiency, pigment content, relative water content-RWC) and antioxidative (catalase-CAT, ascorbate peroxidase-APX, proline) parameters were evaluated in this study using two genotypes of Vicia faba L. differing in drought susceptibility. Drought resistant (Merkur) and sensitive (Piešťanský) genotypes were exposed to 10 days of drought. Activity of CAT and APX of stressed plants rather decreased. Proline content oscillated and no evident trend or significant differences were observed in relation to drought stress. Decrease in RWC was expressed more at susceptible plants, electron-transport chain activity and pigment content were not affected much by drought. Drought susceptible genotype expressed more serious negative effects of drought in morphological parameters, however this genotype was a bit bigger than...
775

Relations entre échanges gazeux foliaires et discrimination isotopique du carbone-13 pendant la photosynthèse : estimations et variations rapides de la conductance mésophyllienne au CO2 / Relationship between carbon isotopic discrimination and leaf gas exchange during photosynthesis : Estimations of mesophyll conductance to CO2

Douthe, Cyril 07 November 2011 (has links)
Les travaux de cette thèse se sont situés autour de la relation entre discrimination isotopique du carbone 13 et échanges gazeux foliaires. Le modèle établi par Farquhar et al. (1982) permet de prédire la discrimination contre le 13C pendant la photosynthèse (delta13C) en tenant compte des processus de diffusion, de carboxylation et décarboxylation engagés pendant la photosynthèse. Cette relation permet d'utiliser delta13C comme indicateur de l'efficience d'utilisation de l'eau (WUE, quantité de carbone fixé en fonction de l'eau consommée), un paramètre particulièrement important dans un contexte de changement climatique, d'agriculture et de sylviculture. Le modèle de delta13C a également été utilisé pour estimer la conductance mésophyllienne au CO2 (gm), un paramètre qui limite fortement la photosynthèse via la disponibilité en carbone dans le chloroplaste. Au cours de nos travaux, nous avons analysé le modèle delta13C pour identifier les paramètres les plus influents dans le modèle, et mis en évidence que l'utilisation du "modèle simple" de delta13C (ignorant gm et les processus de décarboxylation) peut induire un biais important dans l'estimation de WUE. Dans un second temps nous nous sommes concentrés sur les possibles variations à court-terme de gm, un domaine encore sujet a débat. Nous avons confirmé que gm était sensible aux variations de CO2 et d'irradiance sur toutes les espèces d'arbres mesurées dans cette étude. Nous avons aussi montré que ces variations rapides ne peuvent pas être dues a des variations des autres paramètres du modèle, à l'exception possible du paramètre b (discrimination pendant la carboxylation). Nous suggérons que les prochaines études dans ce domaine portent sur (i) la possible variabilité environnementale et génétique du paramètre b et (ii) les mécanismes à l'origine des variations rapides de gm (aquaporines et anhydrases carboniques) / This work was focused on the relationship between isotopic discrimination of 13C during photosynthesis (delta13C) and leaf gas exchange. The model of Farquhar and colleagues (Farquhar et al. 1982) predicts delta13C by accounting for diffusion, carboxylation and decarboxylation processes during the photosynthesis. This relationship is widely used and delta13C is frequently considered as a proxy water use efficiency (WUE, the amount of water required to fix a amount of carbon), an interesting parameter in the context of climate change, crop production and sylviculture. The delta13C model is also used to assess mesophyll conductance to CO2 (gm), that strongly limits photosynthesis via the availability of carbon in the chloroplast. Along this work we analyzed the delta13C model and identified the most important parameters, and highlighted that using the "simple form" of the model (which ignores gm and the decarboxylations) could lead to misestimating WUE. We also focused on the possible rapid variations of gm, a subject still under debate. We confirmed that gm was sensitive to rapid variations of CO2 and irradiance in all species tested in this study. We also showed that apparent rapid variations of gm could not be induced by variations of other parameters in the model, with the exception of parameter b (discrimination during carboxylation). We propose that future studies should focus on (i) the possible environmental and genetic variability of parameter b, and (ii) the physiological processes able to change gm at short time scales (aquaporins and carbonic anhydrase).
776

Synthesis and applications of ruthenium(II)quaterpyridinium complexes and Poly-N-isopropylacrylamide/ acrylic acid copolymers

Siyambalagoda Gamage, Pubudu Hasanka January 1900 (has links)
Doctor of Philosophy / Department of Chemistry / Stefan Bossmann / Tris-homoleptic ruthenium(II)-quaterpyridyl and quaterpyridinium complexes, with +8 and +14 charge were synthesized by utilizing high pressure reaction pathway. These complexes have diameters ranging from 1.82 to 4.55 nm according to the molecular modeling calculations. These ruthenium complexes are highly luminescent and contain long excited state life times. The novel ruthenium(II)-quaterpyridinium complexes exhibit superior reactivity as sensitizer-relay-assemblies (SRA‟s) in sacrificial systems for water and carbon dioxide reductions, while harvesting the ultraviolet- and most of the visible fraction of the incident solar spectrum. Ru(II)-quaterpyridinium complexes and Pd/TiO2 catalysts were successfully used as the catalytic system for the photo catalytic reduction of water and carbon dioxide to hydrogen and methane respectively. Phosphonate-tethered Ru(II)-quaterpyridinium complexes were synthesized from Ru(II)-tris-quaterpyridyl complexes. These complexes form stable adhesive layers on indium tin oxide (ITO) electrodes. A series of differential pulse voltammetry experiments were carried out to measure the ground state and excited state redox potentials of all the Ru(II)quaterpyridinium complexes. The reductive potentials obtained were compared with the reductive potentials of CO2 to CH4 and H2O to H2 reductions. The measurements obtained from the experiments confirmed that it is possible to thermodynamically oxidize water and reduce CO2 by using phosphonate-tethered Ru(II)-quaterpyridinium complexes. These complexes are successfully utilized as prototypes for mycobacterial channel blockers. The Ru(II) complexes show distinct changes in their luminescence spectra when bound to the porin MspA from M. smegmatis, which is a non-pathogenic relative of M. tuberculosis. By using HPLC, we have determined binding constants of the Ru(II)-complexes to MspA in phosphate buffer (0.05 M, pH = 6.8) ranging from 5.2 x 109 M-1 (Ru-C2) to 1.8 x 109 M-1 (Ru-C4). Our findings indicate that channel blocking is a promising treatment strategy for mycobacterial infections. Poly-N-isopropyl-acrylamide/acetic acid copolymers were synthesized and characterized by elemental analysis and gel permeation chromatography. The average composition of the copolymers determined from CHN analysis is in excellent correlation with the feed composition indicating that the radical polymerization process is indeed statistical. Crosslinking of individual polymer chains permitted the generation of ultraflat layers on Mica surfaces by a simple spin-casting procedure, which are able to host the mycobacterial channel protein MspA, while retaining its channel function.
777

Investigation of the regulation of photosynthesis at the molecular level for improvement of plant growth and productivity under limiting light conditions / Investigation of the regulation of photosynthesis at the molecular level for improvement of plant growth and productivity under limiting light conditions

Khuong, Thi thu huong 17 January 2013 (has links)
La lumière est indispensable à la survie des plantes via le processus photosynthétique, pourtant les plantes doivent s'adapter à différentes conditions environnementales où la quantité et la qualité de la lumière peuvent être non optimales pour la photosynthèse. Cela peut provoquer des dégâts photo-induits par formation d'espèces réactives de l'oxygène (ROS), qui sont dangereux pour la plante. Pour limiter la formation des ROS, les plantes mettent en place une régulation importante qui est la dissipation thermique de l'énergie absorbée en excès, appelé Non photochemical quenching (NPQ). Il est connu que la protéine PsbS joue le rôle clé de senseur du pH bas du lumen thylacoïdal, qui est le signal initial pour activer le NPQ. Dans le contexte de cette thèse, on propose d'étudier l'hypothèse que l'absence de la protéine PsbS (diminué NPQ) pourrait augmenter la croissance et la productivité des plantes en conditions contrôlées de faible lumière par l'éminilation de la protéine PsbS chez Arabidopsis thaliana et chez la tomate. Les résultats obtenus indiquent qu'en lumière faible les plantes mutantes montrent une augmentation du rendement de photosystème II conduisant une croissance et un nombre de fleurs significativement augmentés par rapport aux plantes sauvages.De plus, une autre régulation de la photosynthèse, nommée « transitions d'état », est importante pour optimiser la photosynthèse en réponse aux variations de la quantité et de la qualité de la lumière, grâce à la migration réversible des antennes collectrices d'énergie LHCII phosphorylées du PSII au PSI, c'est aussi étudié dans ma thèse. / Light is indispensable for plant survival, but plants have to cope with different environmental situations where light quantity and quality can be not optimal for photosynthesis. This can cause photodamage due to the formation of harmful reactive oxygen species (ROS). To limit ROS formation, plants developed a mechanism important as the dissipation of excess absorbed energy as heat and is called Non Photochemical Quenching (NPQ). The PsbS protein plays the key role of sensor of the low lumenal pH, the signal to activate NPQ. In this thesis, we proposed and investigated the hypothesis that PsbS absence (NPQ decrease) would improve growth under controlled low light upon elimination of the PsbS in Arabidopsis and tomato plants. Results showed that the increase of photosystem II yield in mutant plants leaded to a significant improvement of growth and flower number in mutants as compared with wild type plants under low light, suggesting that this mutation could be useful to improve plant performances in controlled conditions where light is strongly limiting. In addition, another photosynthetic regulation, called “state transitions”, which is important to optimize photosynthesis under variable light for intensity and quality thank to reversible migration of phosphorylated light harvesting complexes LHCII from PSII to PSI also investigated in my thesis.
778

La thioredoxine m4 régule les voies photosynthétiques de transfert cyclique d'électrons autour du photosystème l chez les plantes

Courteille, Agathe 25 January 2013 (has links)
Dans les thylakoïdes des végétaux supérieurs, la réaction photochimique de la photosynthèse implique un transfert d’électrons linéaire ayant lieu entre les photosystèmes II et I et aboutit à la production de pouvoir réducteur (NADPH) et d’ATP. En parallèle à ce transfert linéaire, le transfert cyclique d’électrons autour du PS I court-circuite le PS II et fait intervenir distinctement les complexes PGR (PGR5/PGRL1) et NADPH déshydrogénase (NDH). Ces deux voies alternatives permettent la production d’ATP sans accumulation de NADPH dans le stroma et particulièrement en conditions de stress environnemental. Ces voies sont essentielles pour la physiologie de la plante mais leur régulation demeure mal connue. Une hypothèse a été émise quant à leur régulation redox en étudiant plusieurs thiorédoxines chloroplastiques. Des expériences menées in vivo et in vitro sur des mutants d’Arabidopsis, de tabac et de tomate ont mis en évidence le rôle très spécifique de la thiorédoxine m4 dans la régulation négative des voies dépendantes de PGR et NDH. Une stratégie originale a été mise en place pour capturer les cibles putatives de cette thiorédoxine en mutant le site actif de la protéine chez Arabidopsis. Les résultats obtenus ont montré l’existence de plus de 160 cibles potentielles et certaines d’entre elles pourraient être impliquées dans la régulation des voies dépendantes des complexes PGR et NDH. / In plants thylakoids, the photochemical reaction of photosynthesis implies a linear electron flow between photosystems II and I which produces reducing power (NADPH) and ATP. In addition of linear electron flow, the cyclic electron flow around PS I bypasses PS II and involves distinctly the PGR (PGR5/PGRL1) and the NADPH dehydrogenase (NDH) complexes. These two alternative pathways lead to the production of ATP without accumulation of NADPH in the stroma especially under environmental stress conditions. Both pathways are essential for the plant physiology but their regulation remains unclear. An assumption concerning a redox regulation has been considered by surveying several chloroplastic thioredoxins. In vivo and in vitro studies of Arabidopsis, tobacco and tomato mutants revealed that the m4 thioredoxin plays a very specific role in the down-regulation of the PGR and NDH dependant pathways. An original strategy to capture putative targets of this thioredoxin has been established by mutating the active site of the protein in Arabidopsis. The results showed the existence of more than 160 potential targets and some of them could be involved in the regulation of PGR and NDH dependant pathways.
779

Interceptação de luz, arquitetura e assimilação de carbono em dosséis de capim-xaraés [Brachiaria brizantha (A. Rich.) Stapf. cv. Xaraés] submetidos a estratégias de pastejo rotacionado / Light interception, canopy architecture, and carbon assimilation in Xaraés palisadegrass [Brachiaria brizantha (A. Rich.) Stapf. cv. Xaraés] pastures as affected by rotational grazing strategies

Pedreira, Bruno Carneiro e 03 July 2006 (has links)
A produção de forragem em pastagens é um processo complexo, pois envolve uma série de fatores de ordem fisiológica, morfológica e a interação destes. A partir do conhecimento sobre processos individuais e das interações entre eles pode-se estimar a capacidade assimilatória do dossel usando modelos matemáticos. O objetivo deste trabalho foi avaliar o desempenho agronômico e modelar o potencial fotossintético de dosséis vegetativos de capim-xaraés em função da arquitetura da comunidade vegetal e do ambiente luminoso, em pastagem submetida a estratégias de pastejo intermitente, com freqüências de desfolhação baseadas em interceptação luminosa ou calendário, visando estabelecer uma base racional de manejo e observando o sistema sob o ponto de vista fisiológico da planta e da praticidade do manejo. Além disso, descreve-se as respostas morfo-fisiológicas das plantas e verificando suas relações, com as variações na produção influenciadas pelos tratamentos. O estudo foi realizado na Escola Superior de Agricultura "Luiz de Queiroz", campus da USP localizado em Piracicaba - SP, numa área de Nitossolo Vermelho Eutroférrico, estabelecida com capim Brachiaria brizantha cv. Xaraés, onde foram avaliadas as características agronômicas, morfológicas e fisiológicas deste, tais como produção de forragem, taxas de fotossíntese, índice de área foliar (IAF), interceptação luminosa (IL), ângulos foliares. O delineamento experimental foi inteiramente casualizado com três tratamentos e três repetições, totalizando nove unidades experimentais (piquetes) de 120 m² cada, as quais foram pastejadas por grupos de animais sempre que a IL do dossel chegasse a 95%, ou a 100% ou a cada 28 dias. As estratégias de pastejo apresentaram diferentes acúmulos totais de forragem durante o verão. O tratamento 100% de IL, devido à sua menor freqüência, garantiu à comunidade vegetal um maior intervalo de desfolhações e, conseqüentemente, maior período de crescimento, propiciando maiores acúmulos (22.760 kg MS ha-1) de forragem em relação aos tratamentos 95 % de IL e 28 dias (17.700 kg MS ha-1). Os tratamentos baseados em IL apresentaram alturas de prépastejo, próximas das suas respectivas médias, nas respectivas interceptações luminosas. Isso sugere que a altura pode ser usada como uma ferramenta manejo, o que é confirmado pela sua correlação entre altura e IL para todos os tratamentos (r=0,84). Esta mesma relação ocorre com os valores de IAF (r=0,92), que ao longo do experimento mostrou-se, consistentemente, relacionado aos valores de IL (r=0,92), em todos os tratamentos. No tratamento de maior freqüência (95% de IL) o potencial fotossintético manteve-se máximo do dia médio até o pré-pastejo, devido ao menor intervalo de pastejo e à manutenção de menor área foliar média (2,08) o que propiciou um bom ambiente luminoso, minimizando a competição por luz. Dessa forma, o manejo mais adequado seria aquele que proporciona os maiores valores de fotossíntese de dossel ao longo do ciclo. / Yield formation in pastures is a complex process at it involves factors of both physiological and morphological nature, as well as interactions among them. By understanding individual processes and how they interact it is possible to assess the assimilatory potential of the sward, using through modeling and simulation. The objective of this research was to evaluate the agronomic performance and to model the photosynthetic potential of vegetative swards of Xaraés palisadegrass as a function of canopy architecture and light environment, in pastures under intermittent grazing, where defoliation frequency was dictated either by levels of light interception or by chronological time, in order to try to rationalize management practices under a physiological standpoint and operational practicity. In addition, plant morphological and physiological responses were described in terms of their mutual interplay, as treatments ultimately affected forage yield. The study was conducted at Escola Superior de Agricultura "Luiz de Queiroz", USP campus in Piracicaba, SP, on a kandiudalfic euthrudox using a one-year-old pasture of Xaraés palisadegrass where agronomic, morphological and physiological traits, such as foraqe yield, photosynthesis, leaf area index (LAI), light interception (LI) and leaf angles were studied. The experimental design was completely randomized with three treatments and three replications, for a total of nine experimental units (paddocks) of 120 m² each, which were mob grazed whenever canopy light interception reached 95% or 100%, or every 28 days. Grazing strategies resulted in different seasonal forage yields. The 100% LI treatment made for longer rest periods and less frequent defoliations, resulting in highest (22,760 kg DM ha -1) forage productivity, compared with the 95% LI and the 28-d treatments (mean 17,700 kg DM ha-1). LI-based treatments resulted in less variable pregraze sward heights than the calendar-based treatment, and this suggests that sward height may be successfully used as a management guide. Across treatments sward height was highly correlated with LI (r=0.84) and with LAI (r=0.92) in all treatments, and so were LAI and LI (r= 0.95). Grazed at a higher frequency (95% LI) the canopy responded with maximum levels of assimilation from the midpoint to the end of the rest period, probably due to a lower mean LAI (2.08), which made for a favorable environment and less competition for light. Thus, optimal management seems to be related with higher canopy photosynthesis during the rest period.
780

Aspectos bioquímicos da biossíntese de pigmentos carotenóides em Gonyaulax polyedra (Dinophyceae) / Biochemical aspects of carotenoids biosynthesis in Gonyaulax polyedra (Dinophyceae)

Hollnagel, Heloisa Candia 04 August 2000 (has links)
O dinoflagelado unicelular marinho fotossintetizante Gonyaulax polyedra tem sido utilizado como modelo para o estudo de relógios biológicos. Neste organismo já foram descritos os ritmos de: migração vertical, divisão celular, atividade de superóxido dismutase e nitrato redutase, bioluminescência e capacidade fotossintética. Investigamos a variação circadiana dos pigmentos carotenóides e de RuBisCo II e PCP, as quais estão intimamente ligadas ao processo fotossintético. Experimentos de supressão de espécies reativas de oxigênio (EROs) por carotenóides foram preparados e mostraram que extratos de carotenóides de G. polyedra são capazes de suprimir o O2(1Δg) (oxigênio singlete) in vitro confirmando o importante papel destes no controle das EROs nestas algas. Os extratos metanólicos apresentaram vários pigmentos, tais como clorofila a, β-caroteno e peridinina em diferentes concentrações. A peridinina representa 80 % do total de carotenóides enquanto que o β-caroteno somente 4%. As análises dos cromatogramas de HPLC mostraram que a razão peridinina/clorofila a não varia ao longo de 24 h porém, por outro lado, o β-caroteno apresenta uma variação significativa na sua quantidade, com níveis duas vezes maiores no meio do dia em comparação com os níveis no meio da noite. Esta variação é conservada mesmo quando as células são mantidas em condições de luz constante. A curva de dose-resposta para a síntese de β-caroteno induzida pela luz mostra uma resposta linear com 45 minutos de exposição a luz branca. A indução é máxima quando utilizamos as células do meio período da noite (CT 18) que após esta exposição apresentam níveis de β-caroteno semelhantes as células do meio do dia. Esta alteração de fase no CT 18 sugere que este pigmento pode ser um dos compostos-captadores de luz envolvidos no mecanismo de ajuste de fase por luz em G. polyedra. Culturas de G. polyedra do meio da noite foram expostas à diferentes irradiações (azul, vermelha e verde) e os seus pigmentos extraídos e analisados. Em outra série de experimentos, as células foram mantidas durante o período de claro (12: 12 h) sob diferentes irradiações (vermelha, verde e azul) por 36 horas e os seus pigmentos analisados. Os resultados sugerem que a síntese foto-induzida e a oscilação circadiana do β-caroteno estão ligadas a um fotorreceptor de luz azul/ verde. Nas condições utilizadas não foram observadas variações significativas no conteúdo protéico da RuBisCo II e da PCP ao longo do dia. As análises de RNA total da RuBisCo II mostram que não há variação nos seus níveis quando as células são coletadas no meio do dia e no meio da noite. Quando expostas a condições adversas, G. polyedra apresenta a capacidade de encistar. Embora se conheça bem este mecanismo de defesa, existem poucas informações sobre o estado fisiológico destas células. Células encistadas induzidas por dias curtos apresentam uma alteração na composição de pigmentos com diminuição nas quantidades de β-caroteno e de clorofila a e aumento da quantidade de peridinina, indicando um rearranjo do aparato fotossintético nesta situação, com a peridinina desempenhando um papel mais estrutural. Em consequência, embora o conteúdo protéico de RuBisCo permaneça inalterado, os níveis protéicos de PCP se encontram diminuídos nas células encistadas. / Gonyaulax polyedra, a marine dinoflagellate which has been used as a model to study the biological clock, displays numerous circadian processes, such as bioluminescence, cell aggregation, cell division, superoxide dismutase and nitrate reductase activities and photosynthesis. In this alga, the photosynthesis is maximal in the middle of the day and minimal in the middle of the night. We investigated the pigments content and the amounts of two proteins related to the photosynthesis: ribulose- 1,5- bisphosphate carboxylase/ oxygenase form II (RuBisCo II) and peridinin: chlorophyll a: protein (PCP) in a 24 h cycle. Using the thermal decomposition of 1,4-dimethylnaphtalene endoperoxide, it was shown that the carotenoids could act as effective quenchers of synglet oxygen in G. polyedra. G. polyedra pigments were extracted every three hours over 24 hours. The amounts of peridinin and chlorophyll a remain constant over the day while the levels of β-carotene oscillate, being two times higher at the day than at the night phase. This variation persists when the cells were kept under constant dim light. The dose-response curve for light-induced β-carotene synthesis showed a linear response up to 45 minutes of light exposure, after which night-phase cells contained the same levels of β-carotene as day-phase cells. Cells exposed to light pulses at different times displays the highest β-carotene induction in the middle of the night. This may suggest that β-carotene may be one of the light-harvesting compounds involved in the light induced phase-shift in Gonyaulax polyedra. To identify which was the photoreceptor involved in β-carotene synthesis, cell of the middle of the night-phase (CT 18) were exposed for 45 minutes to different irradiations (red, blue and green) and their pigments extracted and analysed. Also, cells were grown under red, blue and green light during the light phase (12 h light: 12 h dark ) for 36 hours and their pigments analysed. The results suggested that the circadian oscillation and the photoinduced response synthesis of β-carotene, are related to a blue light receptor. The amounts of RuBisCo II and PCP do not change over the circadian cycle when the cultures were grown under constant dim light. The levels of these proteins also remain constant when cells were kept under ither white light or different light qualities (red, blue and green ) in light: dark (12: 12 h) regime. The G. polyedra RuBisCo form II transcrits levels are the same in middle-day and middle-night cells, suggesting a post-translational control for this enzyme in this organism. Adverse environmental conditions elicit the encystment of G. polyedra. Our results showed an alteration in pigment composition of cysts. An increase in peridinin levels and a decrease in β-carotene and chlorophyll a content were observed. Although RuBisCo form II protein levels remained constant, there was a reduction in the amounts of PCP in cysts. This suggests an important role in thylakoids structure stabilizer for free peridinin.

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