O capim tanzânia (Panicum maximum cv. Tanzânia) é promissor para uso em fitorremediação, devido ao seu sistema radicular extenso, boa adaptação a variados ambientes e alto rendimento de biomassa quando bem nutrido. O enxofre faz parte de compostos essenciais ao sistema antioxidante das plantas. A toxicidade causada por alta disponibilidade de metais, como o zinco, no meio de cultivo é um problema ambiental crescente. Objetivou-se avaliar o efeito do enxofre em amenizar o estresse causado pelo excesso de zinco no capim tanzânia. O experimento foi conduzido em casa de vegetação em Piracicaba - SP, durante o verão, em delineamento de blocos casualizados em esquema fatorial fracionado, com as combinações de doses de enxofre (mmol L-1) e zinco (μmol L-1): 0,1/0,7; 0,1/500; 0,1/3000; 1,0/250; 1,0/1000; 1,9/0,7; 1,9/500; 1,9/3000; 2,8/250; 2,8/1000; 3,7/0,7; 3,7/500 e 3,7/3000. Foram avaliadas as seguintes variáveis: componentes de produção (perfilhos, folhas, área foliar, massa de parte aérea e de raízes), concentrações dos nutrientes (N, S, Ca, Mg, Cu, Fe, Mn e Zn), valor SPAD, atividades de enzimas do sistema antioxidante (GR, GPOX, APX, CAT e SOD) e concentração de prolina. A toxicidade de zinco no primeiro crescimento do capim limitou a produção de massa seca e o número total de folhas, enquanto no segundo crescimento também ocorreu limitação na área foliar e no número total de perfilhos. A concentração de enxofre na planta aumentou em função do fornecimento de enxofre e do excesso de zinco no meio de cultivo. O excesso de zinco na planta provocou desequilíbrio na nutrição da planta com os micronutrientes cobre, ferro e manganês. Os indicadores do estresse oxidativo foram sensíveis para detectar a toxicidade do zinco no capim. As reduções na concentração de prolina e nas atividades de enzimas do sistema antioxidante demonstraram o efeito do enxofre em aliviar o estresse por toxicidade de zinco no capim tanzânia. / Tanzania guinea grass (Panicum maximum cv. Tanzania) is promising for use in phytoremediation due to its extensive root system, good adaptation to diverse environments, and high biomass production when well nourished. Sulfur is part of essential compounds to the plant\'s antioxidant system. Toxicity caused by high availability of metals, such as zinc, in the growth medium is a growing environmental problem. The objective was to evaluate the effect of sulfur in mitigating the stress caused by zinc excess in tanzania guinea grass. The experiment was carried out in a greenhouse at Piracicaba, SP, during the summer, in a randomized block design with rates of sulfur (mmol L-1) and zinc (μmol L-1) of 1/0.7; 0.1/500; 0.1/3000; 1.0/250; 1.0/1000; 1.9/0.7; 1.9/500; 1.9/3000; 2.8/250; 2.8/1000; 3.7/0.7; 3.7/500 and 3.7/3000. The response variables evaluated were plant production components (number of tillers, number of leaves, leaf area, and shoots and roots dry matter), nutrient concentrations (N, S, Ca, Mg, Cu, Fe, Mn and Zn), SPAD value, activities of the antioxidant enzymes (GR, GPOX, APX, CAT and SOD) and proline concentration. Zinc toxicity in the first growth of the grass limited the dry mass production and the total number of leaves, while in the second growth also there was limitation in the leaf area and the total number of tillers. The concentration of sulfur in the plant increased as a function of sulfur supply and excess zinc in the growth medium. Excess zinc in the plant caused imbalance in plant nutrition for the micronutrients copper, iron and manganese. The indicators of oxidative stress were sensitive to detect grass toxicity by zinc. Reductions in proline concentration and enzyme activities of the antioxidant system demonstrated the effect of sulfur in alleviating the stress of zinc toxicity in tanzania guinea grass.

A cana-de-açúcar é uma das principais culturas agrícolas no cenário econômico e social brasileiro. Na cultura de cana-de-açúcar o estresse hídrico é o principal fator limitante para o aumento de produtividade, sendo responsável por alterações fisiológicas, bioquímicas e moleculares nas plantas, que podem deflagrar perturbações metabólicas que ativam a morte celular programada (MCP). Sabendo-se que o gene BI-1 apresenta o potencial de reduzir os efeitos da MCP desencadeado por estresses bióticos e abióticos em plantas, este trabalho teve como objetivo analisar plantas transgênicas de cana-de-açúcar que expressam o gene BI-1 de Arabidopsis thaliana (AtBI-1) em condições de estresse hídrico. Também, plantas transgênicas e controle foram inoculadas com o fungo Puccinia melanocephala demonstrando que o processo de transformação genética com o gene AtBI-1 alterou as características pré existentes de resistência a ferrugem marrom nas plantas transgênicas. Os estudos de tolerância ao défict hídrico foram realizados em dois experimentos, o experimento 1 com plantas transgênicas e controles de 90 dias e o experimento 2 com plantas de 60 dias. Plantas do experimento 1 foram analisadas quanto características morfológicas como número de estômatos e tricomas, altura e circunferência do colmo e após ficarem 24 dias sem água foram analisadas quanto a taxa fotossintética, comportamento estomático e conteúdo relativo de água nas folhas, enquanto no experimento 2 as plantas foram analisadas quanto aos teores de prolina, atividades das enzimas guaiacol peroxidase (GPOX), ascorbato peroxidase (APX) e catalase (CAT) após as plantas ficarem 17 dias sob déficit hídrico. Estas enzimas estão envolvidas em processos de desativação de elementos ativos de oxigênio. Os resultados demonstraram que as plantas transgênicas expressando o gene AtBI-1 possuem fenótipo de menor altura, e maior taxa fotossintética, maior comportamento estomático e maior conteúdo relativo de água nas folhas, e assim apresentam maior tolerância ao déficit hídrico que plantas controle. Contudo, houve baixo acúmulo de prolina, baixa atividade da GPOX, APX e CAT nas plantas transgênicas durante o estresse hídrico comparada com as plantas controle do mesmo tratamento. Porém foi observado alta atividade constitutiva da catalase nas plantas transgênicas. A atividade da catalase nestas plantas transgênicas sugere a possibilidade da interação entre AtBI-1 e calmudolinas. Futuros estudos podem contribuir para elucidar se a proteína BI-1 é essencial para a ativação das catalases por calmudolinas. / Sugarcane is one of the main agricultural crops in the Brazilian social and economic scenario. Water stress in the culture of sugarcane is the main limiting factor for increasing productivity accounting for physiological, biochemical and molecular plants that can trigger metabolic disturbances activating programmed cell death (MCP). Knowing that the BI-1 gene has the potential to reduce the effects of MCP triggered by biotic and abiotic stresses in plants, this study aimed to analyze transgenic sugarcane that express the BI-1 gene of Arabidopsis thaliana (AtBI-1) under water stress. Also, transgenic and control plants were inoculated with Puccinia melanocephala fungus demonstrating that the genetic transformation process with the AtBI-1 gene altered the pre-existing characteristics of brown rust resistance in transgenic plants. Studies of tolerance to water deficit were performed in two experiments, the experiment 1 was prepared with transgenic and control plants with 90 days and the experiment 2 used plants with 60 days. Plants from experiment 1 were analyzed as for morphological characteristics such as number of stomata and trichomes, height and diameter of stem after plants being under water for 24 days as were analyzed photosynthetic rate, stomatal behavior, relative water content in leaves while in the experiment 2, plants were analyzed for the levels of proline, enzyme activities of guaiacol peroxidase (GPOX), ascorbate peroxidase (APX) and catalase (CAT) under water deficit for 17 days. These enzymes are involved in deactivation of active elements oxygen. The results demonstrated that the transgenic plants expressing the AtBI-1 gene presented the phenotype of lower height, higher index of leaf area, higher photosynthetic rate, higher stomatal behavior and higher relative water content in leaves than control plants increasing tolerance to drought stress. However, there were low levels of proline, low activity of GPOX activity, APX and CAT in transgenic plants during drought stress compared to control plants of the same treatment, but the observed high constitutive activity of catalase in transgenic plants. Catalase activity in these transgenic plants suggests the possibility of interaction between AtBI-1 and calmudolinas. Future studies may contribute to understand whether the BI-1protein is essential for the activation of catalase by calmudolinas.

Am Beispiel der Wiederkäuer wurde unter Zuhilfenahme von biochemischen und molekularbiologischen Methoden die Adaptation von Pflanzenfressern (Herbivoren) an pflanzliche Sekundärmetabolite wie z.B. Tannine untersucht. Tannine können in nicht an ihren Verzehr adaptierten Spezies durch ihr Proteinbindungsvermögen die Nahrungsverwertung und damit Wachstum und Gesundheit des Pflanzenfressers beeinträchtigen (antinutritive Wirkung). <br /> Einige Wiederkäuerarten wie z.B. das Reh (Capreolus capreolus) haben in ihrem Nahrungsspektrum viele stark tanninhaltige Pflanzen, leiden aber nicht unter den erwähnten postdigestiven Konsequenzen. Eine Möglichkeit, die antinutritive Wirkung von Tanninen zu neutralisieren, besteht in der Produktion tanninbindender Speichelproteine. <br /> Der Speichel verschiedener Wiederkäuerarten wurde auf das Vorhandensein tanninbindender Proteine untersucht. Diese Arten wurden so ausgewählt, dass alle drei Ernährungstypen (Konzentratselektierer, Intermediärtyp, Gras- und Rauhfutterfresser) in den Vergleich eingeschlossen werden konnten. Als Referenzspezies wurde der Konzentratselektierer Reh herangezogen.<br /> Die Speichelproteine des Rehs und die der Intermediärtypen (Rentier, Rangifer tarandus; Damhirsch, Cervus dama; Moschusochse, Ovibos moschatus) banden ungefähr doppelt so effektiv an hydrolysierbare Tannine (Tanninsäure), wie die der untersuchten Gras- und Rauhfutterfresser (Rind, Bos taurus; und Mufflon, Ovis orientalis musimon). Diese Abstufung zeigte sich auch bei der Untersuchung der Bindung an kondensierte Tannine (Quebracho). Eine Ausnahme stellte Mufflonspeichel dar, dieser band ebenso gut an Quebracho wie die Speichelproteine der anderen Ernährungstypen.<br /> Über eine Aminosäuretotalanalyse konnte festgestellt werden, dass der Speichel einiger untersuchter Wiederkäuerarten prolinreiche Proteine (PRPs) enthielt. Unter Ausnutzung ihrer Trichloressigsäure (TCA)-Löslichkeit wurden diese angereichert und genauer untersucht. Die Analyse der TCA-löslichen Speichelproteine der Konzentratselektierer (Reh, Elch) ergab einen relativen Prolingehalt von über 35 %, während beim Moschusochsen noch 29 % gemessen wurden. In Damhirsch- und Rinderspeichel wurden keine prolinreichen Proteine gefunden.<br /> Für die TCA-löslichen Speichelproteine des Rehs konnte eine hohe Tanninbindungskapazität nachgewiesen werden. Diese banden 24 - 30 x effektiver an Tannine als die TCA-löslichen Speichelproteine des Rindes. Die Tanninbindungskapazitäten der TCA-löslichen Speichelproteine von Moschusochse und Damhirsch waren ebenfalls höher als die des Rindes, aber niedriger als die des Rehs. <br /> Die Kohlenhydrat-Analyse der TCA-löslichen Speichelproteine des Rehs erbrachte, dass es sich bei ihnen um Glykoproteine handelt. Mittels Gelfiltration und zweidimensionaler Polyacrylamidgelektrophorese konnten fünf Proteingruppen mit Molekulargewichten zwischen 15 und 50 kd sowie isoelektrischen Punkten zwischen 4,0 und 8,2 detektiert werden. <br /> Von 15 dieser Proteine konnten die N-terminalen Aminosäuresequenzen ermittelt werden. Ausgehend von diesen Informationen wurden Reh-PRP spezifische mRNAs isoliert und partiell sequenziert. Die meisten dieser Fragmente hatten eine gemeinsame 18 Aminosäuren lange C-terminale Sequenz PPPEEQPEE/QSPDEE/DSPSE. <br /> Die Suche nach Übereinstimmungen der analysierten Sequenzen mit anderen Säugetier-PRPs in der Genbank ergab keine sinnvollen Ähnlichkeiten. Die Ergebnisse können zu Informationen über tanninbindende Proteine anderer Wiederkäuer führen. Die Sequenzinformationen stellen einen Ausgangspunkt bei der Analyse der evolutiven Zusammenhänge der Cerviden dar. / Investigation of tannin binding salivary proteins of roe deer and other ruminants: <br /> In this work the adaptation of herbivores to plant secondary metabolites was investigated with help of biochemical and molecular biological methods. In unadapted species plant secondary metabolites as tannins can reduce food digestibility and thus diminish growth rate and health status (antinutritive action). Tannins act through its astringency, that means the high capacity to bind proteins, other macromolecules and metal ions. Some ruminant species feed on tannin containing plant but do not suffer from the mentioned nutritive consequences. The production of tannin binding proteins is one possible adaptation mechanism to neutralize the effects of the tannins.<br /> Saliva of six different ruminant species was investigated for the presence of tannin binding proteins. All three feeding types (concentrate selector, intermediate type and grass and roughage eater) were included in the comparison. <br /> Salivary proteins from roe deer (Capreolus capreolus, concentrate selector) and from the intermediate feeding types (rein deer, Rangifer tarandus; fallow deer, Cervus dama; musk ox, Ovibos moschatus) bound twice as effective to hydrolysable tannins (tannic acid) as those from the investigated grass and roughage eaters (cattle, Bos taurus; moufflon, Ovis orientalis). This differentiation could also be observed investigating the binding capacities to condensed tannins (quebracho) except for moufflon. Moufflon salivary proteins bound with the same intensity to quebracho as the salivary proteins from the other feeding types.<br /> Proline rich proteins (PRPs) could be accumulated from roe deer, moose and musk ox saliva by use of its solubility properties in 5 % trichloro acetic acid (TCA). Roe deer and moose TCA soluble salivary proteins contained more than 35 %, musk ox proteins 29 % proline. In fallow deer and cattle saliva PRPs could not be detected.<br /> A tannin binding assay demonstrated for the TCA soluble salivary proteins from roe deer, musk ox and fallow deer but not from cattle, that they are able to bind tannins. Roe deer salivary proteins bound 24 to 30 more effective to tannins as cattle proteins. Tannin binding capacity of the proteins from musk ox and fallow deer saliva was higher as those from cattle but lower as those from roe deer.<br /> For further analysis of ruminant tannin binding proteins we chose roe deer as reference species. Carbohydrate analysis of TCA soluble proteins from roe deer saliva showed that they were glycoproteins. With help of gel filtration and two dimensional polyacrylamid gel electrophoresis five proteins groups with molecular weights from 15 to 50 kd and isoelectric points from 4.0 to 8.2 could be detected.<br /> N-terminal amino acid sequences of 15 of the roe deer salivary TCA soluble proteins were determined by Edmann degradation. This information led to partially sequenced roe deer PRP specific cDNA. An 18 amino acid long C-terminal sequence was common in most of the clones. The obtained roe deer PRP sequences did not match with known mammalian PRP sequences from data banks. <br /> The finding in this work can lead to information about salivary tannin binding proteins in other ruminants. The sequence information represent a starting-point for the investigation of cervid evolution.

Magistrantūros studijų darbe pateikiami egzogeninio prolino poveikio žaliosios šerytės augimui ir vystymuisi tyrimų rezultatai. Darbo objektas – žalioji šerytė (Setaria viridis (L.) P. Beauv.). Darbo metodai: IV organogenezės tarpsnyje žaliosios šerytės augalai nupurkšti prolino tirpalais ir auginti programuojamoje auginimo kameroje esant 20/18 °C (diena/naktis) temperatūrai, 16/8 val. (diena/naktis) fotoperiodui, 50 µmol m-2 s-1 apšviestumui. Fotosintezės pigmentai, augalo žalia masė ir sausos medžiagos kiekis nustatyti praėjus 1, 5, 10 ir 15 dienų po purškimo. Esant 35-40% substrato drėgniui augalai nupurkšti 10 mM L-prolino tirpalu ir toliau auginti programuojamoje auginimo kameroje sausros sąlygomis. Fotosintezės pigmentai, augalo žalia masė ir sausos medžiagos kiekis nustatyti po 3 ir 6 sausros dienų. Darbo rezultatai. Žaliosios šerytės augalai sukaupė statistiškai patikimai didesnį chlorofilo a ir karotenoidų kiekį visų tirtų L-prolino koncentracijų poveikyje. Augalus paauginus 10 ir 15 dienų po purškimo nustatytas neigiamas egzogeninio L-prolino poveikis chlorofilo b kiekiui žaliosios šerytės augaluose. Neigiamas egzogeninio L-prolino poveikis žaliai masei nustatytas praėjus 5 ir 10 dienų po purškimo, o paauginus augalus 15 dienų neigiamas L-prolino poveikis augalo žaliai masei beveik eliminavosi. Visos tirtos L-prolino koncentracijos praėjus 15 dienų po purškimo didino sausos medžiagos kiekį žaliosios šerytės augaluose ir spartino generatyvinių organų susidarymą... [toliau žr. visą tekstą] / The objective of this study was to investigate the effect of exogenous proline on growth and development of green millet plants. Object of the work – green millet (Setaria viridis (L.) P. Beauv.). Methods of the work: green millet plants were to spray with proline and growing in growth chamber under 20/18 °C (day/night) temperature, 16/8 h (day/night) photoperiod, 50 µmol m-2 s-1 light intensity. Amount of photosynthetic pigments, fresh mass of plants, dry weight was evaluated after 1, 5, 10 and 15 days after treatment. When substrate humidity was 35-40%, plants were to spray with 10 mM proline and growing in growth chamber under moisture deficit. The results of work. Green millet plants accumulated significantly higher chlorophyll a and carotenoids contents under influence of all proline concentration tested. Negative effect of exogenous L-proline on amounts of chlorophyll b in green millet plants has been obtained after 10 and 15 growing days. Negative effect of exogenous L-proline on fresh mass of green millet plants has been obtained after 5 and 10 growing days, however this effect was almost eliminated after 15 growing days. All tested L-proline concentrations increased dry weight after 15 growing days and accelerated formation of generative organs. After 3 and 6 drought stress treatment, exogenous proline didn’t have any effect on chlorophyll b content, but resulted in increasing of chlorophyll a content. Exogenous proline didn’t have significant effect on dry weight... [to full text]

The innate immune pathways that stimulate the expression of cytokines and proapoptotic factors in response to infection are triggered by the activation of the cytosolic receptors retinoic acid-inducible gene I (RIG-I) and melanoma differentiationassociated gene 5 (MDA5). Activation of both receptors occurs as a result of binding to RNA. MDA5 only recognizes double stranded forms of RNA, whereas RIG-I is capable of recognizing both single and double stranded RNA. In vivo, MDA5 is known to be stimulated by long (>1 kb) strands of RNA, forming filaments along the phosphate backbone. However, the manner in which MDA5 can recognize the terminal end of its RNA ligand is uncertain. I have examined the mechanism of binding of the MDA5 protein by comparing MDA5 binding to short (<18 bp) blunt RNA, 5’ triphosphate RNA, and RNA with a 3’ or 5’ overhang. It is shown that while the MDA5 protein regulatory domain (RD) is essential for RNA recognition, the MDA5 RD only weakly recognizes short double stranded RNA ligands with overhangs or a 5’ triphosphate group. The Cys951 residue was shown to disrupt stability of the MDA5 RD-RNA complex. Binding analyses were performed using a combination of SDS-PAGE, gel filtration analysis, and nondenaturing gel electrophoresis. In addition, structural data was gathered by crystallization of the MDA5 RD-RNA complex using X-ray crystallography. These results help to establish the manner in which MDA5 is regulated predominantly to the binding of long RNA ligands. Also included in this document is structural data on the dimer form of the PepQ protein from E. coli. PepQ is a highly conserved proline peptidase that has a secondary activity of hydrolyzing organophosphorus triesters, toxic compounds found in many pesticides. The PepQ protein was crystallized and analyzed by X-ray diffraction. The dimer interface was clearly defined within the structure and provides insight into how the active dimer forms from the PepQ monomer.

Thesis (Ph. D.)--University of Alabama at Birmingham, 2007. / Title from first page of PDF file (viewed Feb 17, 2009). Includes bibliographical references.

No presente trabalho desenvolveu-se uma rota sintética em fase sólida para obter de forma rápida o esqueleto da hiacintacina 110 utilizando-se L-Boc-prolina 101 como bloco de construção. Não há relatos na literatura, até o momento, do uso da Lprolina como material de partida para a síntese de núcleos pirrolizidínicos, entretanto o estudo desta rota é uma alternativa interessante porque possibilita a síntese destes biciclos em apenas 4 etapas. A síntese iniciou com o acoplamento entre a L-Bocprolina 101 e a resina Merifield 41 em condições clássicas. Após a retirada do grupamento protetor com ácido cloridríco por 40 minutos em temperatura ambiente, procedeu-se à adição de Michael do produto 103 ao propiolato de etila 116 gerando o enaminodiéster 104. Na última etapa usou-se DBU sob irradição de microondas por 10 minutos, obtendo-se o azabiciclo 110 com redimento de 37%. Em solução, através da adição de Michael entre a L-prolina O-protegida 95 e alcinos e alceno ativados obtevese adutos cuja estereoquímica das olefinas foi sempre E, com bons rendimentos (60- 95%). Os enaminodiésteres 97 e 118 foram facilmente convertidos nos seus correspondentes aminodiésteres 124 e 126 com NaBH(OAc)3 com redimento de 65%. Na etapa da ciclização de Dieckmann obteve-se sucesso utilizando Na/THF/3h sob refluxo gerando o azabiciclo[3.3.0]octano desejado com rendimento de 50-60%. Os análogos das hiacintacinas foram sintetizados a partir da redução dos produtos 109, 132 e 133 com LiAlH4/THF e rendimentos de 52%. Os produtos 109, 132, 133, 135, 136 e 137 foram testados para atividades antiparasitárias e apresentaram bons resultados para a ação carrapaticida com doses até 0,625μg/ml em ovos e 1,25μg/ml em larvas. / We present here an original solid-phase approach to synthesize rapidly the hyacinthacine core 110 using L-proline N-Boc 101 as building block. There is no reference in the literature wich uses L-proline as building block to synthesize this pirrolizidine skeleton. The study of this possibility would be extremely interesting because it could be used to prepare structural systems related to hyacinthacine in only four steps. The synthesis initiates with the coupling of L-Boc-proline 101 to Merrifield resin 41 under classical conditions. The protection group was removed with hidrogen chloride and the Michael addition was proveeded with adduct 103 and ethyl propiolate 116, afording 104 in satisfactory yields. At least, the azabicycle 110 was obtained by reacting 98 using DBU as catalyst at MW irradiation for 10 min, in good yields (37 %). In solution, we have proceed to Michael addition of L-proline O-protected 95 to activated alkynoates/alkenes conducing to aducts where the stereochemistry of the olefins was always E, with excellent yield (60-95%). Enaminoesters 97 and 118 could be easily converted into the corresponding aminodiesters 124 and 126 by NaBH(OAc)3 in 65% yield. In the step of the Diekmann cyclization of aminodiesters the sistem Na/THF/3h at reflux afforded the derived azabicyclo[3.3.0]octanes in 50- 60% yield. The analogs of the hyacinthacines were synthesized with reduction of 109, 132 and 133 with LiAlH4/THF in 52% yield with the stereochemistry to be determined. The products 109, 132, 133, 135, 136 and 137 were tested for parasitic activity and showed good results. Compounds 109, 132, 135 and 136 killed 100% of the larvae at the concentrations of 5 μg/ml and 2.5 μg/ml being not toxic at the lower concentrations. Compounds 133 and 137 were more toxic being lethal to the larvae at concentrations of 1.25 μg/ml but did not case death at the lower concentrations.

Made available in DSpace on 2014-06-11T19:26:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-10Bitstream added on 2014-06-13T20:15:26Z : No. of bitstreams: 1 marques_dj_me_botfca.pdf: 1236525 bytes, checksum: b3e82023e001449560012288e4ba0ba8 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A cultura da berinjela vem se destacando muito, principalmente na Europa, Estados Unidos e também no Brasil, com aumento da demanda por hortaliças. Dentre os fertilizantes potássios mais utilizados no Brasil, o cloreto de potássio, é responsável por 95% do consumo. Entre os fatores que afetam a resposta à adubação potássica, destacam-se salinidade e acidez do solo. O experimento foi divido em duas fases, sendo a primeira desenvolvida no Departamento de Produção Vegetal – Horticultura – UNESP, Campus de Botucatu, São Paulo. A segunda fase, compreendendo as análises bioquímicas, foram conduzidas no Departamento de Química e Bioquímica do Instituto de Biociências - UNESP, Campus de Botucatu, São Paulo. Utilizou-se a variedade de berinjela denominada de Embu, adotando-se delineamento em blocos casualizados, em esquema fatorial 2 x 4, com cinco repetições, com duas fontes de potássio (cloreto e sulfato de potássio) e quatro doses crescentes dos fertilizantes (250, 500, 750 e 1000 kg K2O ha-1) Observou-se que as plantas cultivadas com doses elevadas de ambas as fontes potássicas, induziram maior atividade das enzimas superoxido dismutase e catalase, além do aumento da concentração de L-prolina. Estas alterações indicam que estes parâmetros podem ser adotados como indicadores de estresse mineral na cultura da berinjela. Além dos fatores bioquímicos, observou-se que as variações das fontes potássicas influenciaram nos parâmetros biométricos e agronômicos da berinjela, notadamente quando aplicados em excesso. Avaliando-se os resultados do experimento, concluiu-se que a fonte KCl apresentou efeito salino superior, quando comparado com a fonte K2SO4 baseado no efeito mais pronunciado deste adubo, os parâmetros avaliados. Para ambas as fontes potássicas aplicadas, observou-se que o excesso de potássio afetou os parâmetros biométricos, produção e atividade enzimática das plantas. / Eggplant cultivation has extensively developed, mainly in Europe, United States and Brazil due to the increase in the demand for vegetables. Of the potassium fertilizers most frequently used in Brazil, potassium chloride satisfies 95% of the needs. Among the factors affecting the response to potassium fertilization are soil salinity and acidity. This experiment was divided into two steps: the first was conducted in the facilities of the Department of Plant Production, Horticulture, São Paulo State University – UNESP, Botucatu Campus, São Paulo State, Brazil. The second step which consisted of biochemical analyses was carried out in the Department of Chemistry and Biochemistry, Institute of Biosciences, UNESP, Botucatu Campus. The eggplant variety Embu was used, and the experimental design was in randomized blocks, 2 x 4 factorial arrangement, with five replicates, including 2 potassium sources (potassium chloride and sulfate) and four increasing doses of the fertilizers (250, 500, 750, and 1000 kg K2O ha-1). Elevated doses of both potassium sources led to higher activity of the enzymes superoxide dismutase and catalase in plants, as well as to an increasing L-proline concentration. Based on such alterations, those parameters can be used as indicators of mineral stress in eggplant culture. Besides biochemical factors, biometric and agronomical parameters were also influenced by variations in the potassium sources, mainly when applied in excess. The present experiment demonstrated that KCl source had higher saline effect, compared with K2SO4 source, considering the higher effect of the former on the evaluated parameters. As regards both potassium sources, the excess of potassium affected biometric parameters, yield and enzymatic activity.

No presente trabalho desenvolveu-se uma rota sintética em fase sólida para obter de forma rápida o esqueleto da hiacintacina 110 utilizando-se L-Boc-prolina 101 como bloco de construção. Não há relatos na literatura, até o momento, do uso da Lprolina como material de partida para a síntese de núcleos pirrolizidínicos, entretanto o estudo desta rota é uma alternativa interessante porque possibilita a síntese destes biciclos em apenas 4 etapas. A síntese iniciou com o acoplamento entre a L-Bocprolina 101 e a resina Merifield 41 em condições clássicas. Após a retirada do grupamento protetor com ácido cloridríco por 40 minutos em temperatura ambiente, procedeu-se à adição de Michael do produto 103 ao propiolato de etila 116 gerando o enaminodiéster 104. Na última etapa usou-se DBU sob irradição de microondas por 10 minutos, obtendo-se o azabiciclo 110 com redimento de 37%. Em solução, através da adição de Michael entre a L-prolina O-protegida 95 e alcinos e alceno ativados obtevese adutos cuja estereoquímica das olefinas foi sempre E, com bons rendimentos (60- 95%). Os enaminodiésteres 97 e 118 foram facilmente convertidos nos seus correspondentes aminodiésteres 124 e 126 com NaBH(OAc)3 com redimento de 65%. Na etapa da ciclização de Dieckmann obteve-se sucesso utilizando Na/THF/3h sob refluxo gerando o azabiciclo[3.3.0]octano desejado com rendimento de 50-60%. Os análogos das hiacintacinas foram sintetizados a partir da redução dos produtos 109, 132 e 133 com LiAlH4/THF e rendimentos de 52%. Os produtos 109, 132, 133, 135, 136 e 137 foram testados para atividades antiparasitárias e apresentaram bons resultados para a ação carrapaticida com doses até 0,625μg/ml em ovos e 1,25μg/ml em larvas. / We present here an original solid-phase approach to synthesize rapidly the hyacinthacine core 110 using L-proline N-Boc 101 as building block. There is no reference in the literature wich uses L-proline as building block to synthesize this pirrolizidine skeleton. The study of this possibility would be extremely interesting because it could be used to prepare structural systems related to hyacinthacine in only four steps. The synthesis initiates with the coupling of L-Boc-proline 101 to Merrifield resin 41 under classical conditions. The protection group was removed with hidrogen chloride and the Michael addition was proveeded with adduct 103 and ethyl propiolate 116, afording 104 in satisfactory yields. At least, the azabicycle 110 was obtained by reacting 98 using DBU as catalyst at MW irradiation for 10 min, in good yields (37 %). In solution, we have proceed to Michael addition of L-proline O-protected 95 to activated alkynoates/alkenes conducing to aducts where the stereochemistry of the olefins was always E, with excellent yield (60-95%). Enaminoesters 97 and 118 could be easily converted into the corresponding aminodiesters 124 and 126 by NaBH(OAc)3 in 65% yield. In the step of the Diekmann cyclization of aminodiesters the sistem Na/THF/3h at reflux afforded the derived azabicyclo[3.3.0]octanes in 50- 60% yield. The analogs of the hyacinthacines were synthesized with reduction of 109, 132 and 133 with LiAlH4/THF in 52% yield with the stereochemistry to be determined. The products 109, 132, 133, 135, 136 and 137 were tested for parasitic activity and showed good results. Compounds 109, 132, 135 and 136 killed 100% of the larvae at the concentrations of 5 μg/ml and 2.5 μg/ml being not toxic at the lower concentrations. Compounds 133 and 137 were more toxic being lethal to the larvae at concentrations of 1.25 μg/ml but did not case death at the lower concentrations.

Made available in DSpace on 2016-12-08T16:44:34Z (GMT). No. of bitstreams: 1 PGPV09MA042.pdf: 353135 bytes, checksum: f6e276c764d17f2d788c1c6b5cd1e088 (MD5) Previous issue date: 2009-02-05 / The objective of this study was to evaluate the tolerance of Eucalyptus dunnii Maiden, E. benthamii Maiden & Cambage, E. grandis W. Hill ex Maiden e E. saligna Sm., to low temperature (LT&#8325;&#8320;) of seedings when submitted to diferente rustification periods, using as cold tolerance indicators the contentes of total soluble carbohydrates and proline in the leaves. The work was carried out in the Plant Physiology Laboratory of the Agricultural and Veterinary Sciences Center of the University of Santa Catarona State UDESC. Two experiments were performed using a randomized block design. On the first experimente, (clone K-1007) Eucalyptus dunnii seedlings were submitted to diferente periods rustification (0,7,21 and 42 days) and after each rustification period, the seedling were exposed to diferente sub-zero temperatures (-2°, -4°, -6° and -8°C). On the second experimente, seedlings of species Eucalyprtus dunnii (clone KF 09), E. benthamii (clone ARV 116), E. grandis (clone 360 ) and E. saligna (clone 2864) were submitted to two rustification periods (0 and 21 days) and after each rustification period, the seedlings were exposed to three sub-zero temperatures (-2°, -5° and -8°C). In the both experiments the rustification was performed under a regime of daily and night temperatures of 1°C and 5°C, respectively, with photoperiod of 12h. were evaluated the concentrations of total soluble carvohydrates and proline in the leaves, the LT&#8325;&#8320;, the electric conductivity, and the damage index. The data were submitted to analysis of variance followed by the linear regression analysis, Pearson s correlation and in the experimente 2, as well as, to the Test of Tukey (p<0.05). the data obtained on the first experimente with E. dunnii, showed na invrease in the tolerance of this species to low temperatures with increase of leaf contentes of total soluble carbohydrates when submittedmto a rustification period up to 35 days. It was also observed that there is a relationship between the cold tolerance and the increase of leaf total soluble carbohydrates. On the second experimente, diferences were observed between the species regarding to the cold tolerance. The E. dunnii and E. benthamii presented a higher cold tolerance followed by the E. saligna and E. grandis. However, in all the species studied, it was observed a good relationship between to cold tolertance (lower LT&#8325;&#8320;) with the rustification period and that tolerance was correlated to the increase in the leaf contentes of total soluble carbohydrates. The presence of proline in the leaf tissues was found only in the plants of E. dunnii on first experimente and there was no increase within the rustification period. In both experiments it was not observed any relationship between the leaf proline contente and cold tolerance. It was concluded that the increase of cold tolerance of eucalyptus species, in response to low temperature of the plants, was mostrly due to the increase of total soluble carbohydrates in leaf contentes, rather than to the leaf proline contents / Objetivou-se com este trabalho avaliar a tolerância à baixa temperatura (LT50) em mudas das espécies de Eucalyptus dunnii Maiden, E. benthamii Maiden & Cambage, E. grandis W. Hill ex Maiden e E. saligna Sm., submetidas a diferentes períodos de rustificação, usando como indicadores de tolerância ao frio os teores foliares de carboidratos solúveis totais e prolina. Foram realizados dois experimentos, usando-se o delineamento experimental blocos ao acaso, no Laboratório de Fisiologia Vegetal do Centro de Ciências Agroveterinárias da Universidade do Estado de Santa Catarina UDESC. No primeiro experimento, mudas de Eucalyptus dunnii (clone K-1007) foram submetidas a diferentes períodos de rustificação (0, 7, 21 e 42 dias) e após cada período de rustificação as mudas foram expostas a quatro gradientes de temperatura negativa (-2,-4,-6 e -8ºC). No segundo experimento, mudas das espécies de E. dunnii (clone KF 09), E. benthamii (clone ARV 116), E. grandis (clone 360) e E. saligna (clone 2864) foram submetidos a dois períodos de rustificação (0 e 21 dias), sendo que após cada período de rustificação as mudas foram expostas a três gradientes de temperatura negativa (-2,-5 e -8ºC). Em ambos os experimentos a rustificação foi realizada sob um regime de temperaturas diurnas e noturnas de 1ºC e 5ºC, respectivamente, com fotoperíodo de 12 horas. Foram avaliadas as concentrações foliares de carboidratos solúveis totais e de prolina, o LT50, através da condutividade elétrica, e o índice de dano. Os dados foram submetidos à análise da variância e após a análise de regressão linear, teste de correlação de Pearson e, no experimento 2, também ao teste de Tukey (p<0,05). Os dados obtidos no primeiro experimento, com E. dunnii, demonstram que ocorreu um aumento na tolerância desta espécie a baixas temperaturas observando-se um incremento nos teores foliares de carboidratos solúveis totais até 35 dias de rustificação. Também foi observado que existe uma relação entre a tolerância ao frio e o incremento nos teores foliares de carboidratos solúveis totais. No segundo experimento, houve diferenças entre espécies quanto à tolerância ao frio, sendo que as espécies E. dunnii e E. benthamii apresentaram maior tolerância ao frio seguido das espécies E. saligna e E. grandis. No entanto, em todas as espécies estudadas foi observada uma maior tolerância ao frio (menor LT50) das plantas submetidas à rustificação e que esta tolerância está associada ao incremento nos teores foliares de carboidratos solúveis totais. A presença de prolina nos tecidos foliares foi identificada apenas nas plantas de E. dunnii do primeiro experimento e não houve alteração na concentração com o aumento do período de rustificação. Em ambos os experimentos não foi observada correlação entre os teores foliaresde prolina com a tolerância ao frio. Conclui- e que o aumento da tolerância ao frio de espécies de Eucalyptus, em resposta à rustificação das plantas, deve-se em grande parte ao incremento nos teores foliares de carboidratos solúveis totais e não aos teores foliares de prolina