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1
Andrews, Christopher John. "Glutathione transferases in soybean Glycine max (L.) Merr." Thesis, Durham University, 1999. http://etheses.dur.ac.uk/4857/.
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Glutathione transferases, also known as Glutathione S-transferases (GSTs), are a diverse group of enzymes that catalyse the conjugation of the tri-peptide glutathione to a wide range of electrophilic substrates. Their biological function in endogenous metabolism in plants is not well characterised, although their role in herbicide metabolism and herbicide selectivity is well documented. Many herbicides used in soybean. Glycine max (L.) Merr., are selective against weeds due to their rapid detoxification in the crop through conjugation with homoglutathione (γ-glu-cys-β-ala), the predominant free thiol in many legumes. However, an in depth characterisation of the GSTs which can potentially catalyse these reactions in soybean has never been performed. This work describes the biochemical and molecular characterisation of GSTs in soybean with emphasis on the identification of specific isoenzymes involved in herbicide metabolism. GST activity toward the chloroacetanilide herbicides acetochlor and metolachlor, the diphenyl ethers acifluorfen and fomesafen and the sulphonyl urea chlorimuron-ethyl were all detected in crude protein extracts from five-day-old suspension cultured soybean cells. GST activity was also determined in five-day-old soybean seedlings, though this activity was significantly lower than that observed with the cell suspension cultures. Treatment of soybean plants with herbicides and herbicide safeners resulted in increased GST activity toward the model substrate l-chloro-2,4-dinitrobenzene (CDNB), but no change in activity toward herbicide substrates. In both plant and cell cultures GST-catalysed conjugation of the diphenyl ethers acifluorfen and fomesafen was over five-fold greater in the presence of homoglutathione as compared with glutathione. The preferential detoxification of these herbicides in the presence of homoglutathione appeared to be an important determinant of their rapid detoxification in soybean and an important factor in herbicide selectivity. GSTs were purified from five-day-old soybean cell cultures using S-hexylglutathione affinity chromatography and anion-exchange chromatography. A combination of reversed-phase HPLC, SDS-PAGE and MALDI-TOF mass spectrometry of the purified fractions indicated the presence of nine putative GST subunits, each with a molecular mass between 25 and 29 kDa. Soybean GST cDNA clones were obtained using a combination of RT-PCR, utilising degenerate oligonucleotides designed to conserved regions within plant GSTs, and screening of cDNA libraries prepared from soybean plants and cell cultures. This process failed to identify any theta-type GSTs, the class associated with herbicide detoxification in maize. In contrast, seven distinct tau-type GSTs were isolated together with a number of clones showing minor variations in individual sequences. Expression of these cDNAs in Escherichia coli showed the purified recombinant GSTs were active toward a diverse range of substrates, and possessed additional glutathione peroxidase activity. GST activities for each recombinant enzyme varied with substrate and thiol type, with a marked preference for homoglutathione with selected substrates. From the work reported in this study it would appear that the tau-type GSTs of soybean are at least as complex as those previously reported in cereals and have an important role in determining herbicide metabolism and selectivity in this major crop.
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Chave, Karen Judy. "Analysis of variant cytosolic serine hydroxymethyltransferases." Thesis, University of Surrey, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.336746.
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Grove, Geraldine. "Variability in the measurement of protein turnover in man using the end product method and '1'5N glycine." Thesis, University of Southampton, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.240627.
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Chronis, Demosthenis. "Sulfur metabolism in Glycine max [L.] Merr characterization of serine acetyletransferase and O-acetylserine (thiol) lyase /." Diss., Columbia, Mo. : University of Missouri-Columbia, 2006. http://hdl.handle.net/10355/4483.
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Thesis (Ph.D.)--University of Missouri-Columbia, 2006.The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on May 1, 2009) Vita. Includes bibliographical references.
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Souza, Karine Bresolin de. "Efeitos da mal nutrição protéica sobre o metabolismo da glicina em cerebelo de ratos durante o seu desenvolvimento." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2003. http://hdl.handle.net/10183/2684.
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Resumo não disponível.Malnutrition is a worldwide problem affecting millions of unborn and young children during the most vulnerable stages of brain development (1). All restriction of protein during the perinatal period of life can alter the development of mammalian fetus and have marked repercussions on development of the Central Nervous System (CNS). The brain is vulnerable to protein malnutrition with altered morphologic and biochemical maturation, leading to impaired functions. The focus of this study is to investigate [U-14C]glycine metabolism in malnourished rats submitted to pre- and postnatal protein deprivation (diet: 8% protein with addition and without addition of L-methionine) on glycine metabolism of rats (normonourished group: 25% protein). It was observed that protein malnutrition alters oxidation to CO2, conversion to lipids and protein synthesis from [U-14C]glycine in cerebellum of malnourished rats without addition of L-methionine on a diet at 7 and 21 days of postnatal life. Our results also indicate that protein malnutrition causes a retardation in the normally ordered progression of brain development, and the malnourished groups have smaller cells, reduction in cell numbers and smaller cerebellar weight comparing to the control group.
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Picking, Jonathan William. "Glycine Betaine and Proline Betaine Specific Methyltransferases of the MttB Superfamily." The Ohio State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=osu1563468258124346.
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Sutherland, Steven Thomas. "Studies on the metabolism of oxalate, glyoxylate, glycolate and glycine by peroxisomes and mitochondria from rat liver /." The Ohio State University, 1991. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487693923196163.
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Wylde, Elinor. "Drug metabolism and pharmacokinetics in the lead optimisation of novel positive allosteric modulators of α1 strychnine sensitive glycine receptors." Thesis, University of Liverpool, 2015. http://livrepository.liverpool.ac.uk/2048099/.
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Chronic pain is a condition that is thought to affect roughly 8 million people in the UK. It is classified as pain that persists for more than 6 months. Chronic pain is commonly associated with depression, insomnia, anxiety and poor quality of life. Many treatments for chronic pain are accompanied by numerous debilitating side-effects, this in combination with insufficient pain relief means that approximately 50% of patients will discontinue their treatment. Most sufferers choose to live with the pain rather than deal with numerous adverse-effects. There is a great need for new therapeutics that are specifically designed to target the underlying mechanisms of chronic pain, therefore providing safer and more effective treatments. One such mechanism is the down-regulation of strychnine-sensitive glycine receptors (SSGRs) localised in the dorsal horn. Glycinergic activity is known to be inhibitory and artificial stimulation can produce analgesia. Positive allosteric modulators acting on α1 SSGRs may able to compensate for the inhibitory glycinergic activity that is reduced in chronic pain. Previous work within the group lead into the identification of propofol analogues designed to be novel positive allosteric modulators of α1 SSGRs. Work presented in this thesis describes the generation and optimisation of these analogues with a focus of drug metabolism and pharmacokinetics. The hit to lead process has resulted in the development of a lead compound that is highly potent at the target, has excellent pharmacokinetic and safety profiles and is able to produce high levels of analgesia in an animal model of neuropathic pain.
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Creighbaum, Adam J. "Examination and reconstitution of the glycine betaine-dependent methanogenesis pathway from the obligate methylotrophic methanogen Methanolobus vulcani B1d." Miami University / OhioLINK, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=miami158754951585964.
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Witt, Joshua. "The Glycine and Proline Reductase Systems: An Evolutionary Perspective and Presence in Enterobacteriaceae." Honors in the Major Thesis, University of Central Florida, 2013. http://digital.library.ucf.edu/cdm/ref/collection/ETH/id/1656.
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The Glycine and Proline Reduction systems are two of the best characterized selenoenzymes in bacteria and have been found to occur in a wide variety of clostridia [1-5]. These enzymes are utilized to reduce glycine or D-proline to obtain energy via substrate level phosporylation or membrane gradients, respectively [6, 7]. This includes the pathogens C. difficile and C. botulinum [5, 8]. Strains of C. difficile are activate toxigenic pathways whenever either of these pathways is active within the cell [5, 8]. Though evolutionary studies have been conducted on ammonia producing bacteria [9] none has been done to directly characterize these two system by themselves. This includes an understanding of whether or not this system is transferred between organisms, as many of the clostridia that are to be studied are known to have an “open genome.” [8, 10] With this information we were able to generate a phylogenic model of the proline and glycine reduction systems. Through this analysis, we were able to account for many clostridial organisms that contain the system, but also many other organisms as well. These included enterobacteriaceae including a strain of the model organism, Escherichia coli. It was further concluded that Glycine Reductase was a much less centralized system and included a wide range of taxa while Proline Reductase was much more centralized to being within the phyla of firmicutes. It was also concluded that the strain of E. coli has a fully functional operon for Glycine Reductase.B.S.
Bachelors
Burnett School of Biomedical Sciences
Molecular Biology and Microbiology
11
Han, Ling. "Physiology of Escherichia coli in batch and fed-batch cultures with special emphasis on amino acid and glucose metabolism." Doctoral thesis, KTH, Biotechnology, 2002. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-3334.
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The objective of this work is to better understand themetabolism and physiology ofEscherichiacoli(W3110) in defined medium cultures with thelong-term goal of improving cell yield and recombinant proteinproductivity.
The order of amino acid utilization inE. colibatch cultures was investigated in a medium with16 amino acids and glucose. Ser, Pro, Asp, Gly, Thr, Glu andAla were rapidly consumed and depleted at the end of theexponential phase, while His, Arg, Val, Met, Ile, Leu, Phe, Lysand Tyr were consumed slowly during the following linear growthphase. The uptake order correlated to the maximum specificconsumption rate. Of the rapidly consumed amino acids onlyglyine and threonine improved growth when added individually.Serine was the first amino acid to be consumed, but inhibitedglucose uptake initially, which presumably is related to thefunction of PTS. Valine inhibited cell growth could be releasedby isoleucine. The critical medium concentration of valinetoxicity was 1.5 - 3 µmol L-1. Valine uptake was associated with exchange ofisoleucine out of the cells.
Glycine significantly increased the cell yield,Yx/s,and growth rate ofE. coliin batch cultures in a glucose-mineral medium.Maximum effect occurred at pH 6.8, at 6 - 12 mmol L-1glycine, and below 1.15 g dw L-1.13C NMR technique was employed to identify [1-13C], [2-13C]and [1,2-13C]acetate in the cultures supplied with [2-13C]glycine. The NMR data revealed that littledegradation of added glycine occurred, and that serine/glycinebiosynthesis was repressed below 1.15 g dw L-1, implicating that glycine was a source ofglycine, serine, one-carbon units, and threonine. Above 1.15 gdw L-1, 53% of the consumed glycine carbon was excretedas acetate. Degradation of glycine was associated with anincreased uptake rate, cleavage by GCV, and degradation of bothglycine- and glucose-derived serine to pyruvate. This switch inmetabolism appears to be regulated by quorum sensing.
A cell density-dependent metabolic switch occurred also inthe central metabolism. A 2 - 3 fold decrease in mostglycolytic and TCA cycle metabolites, but an increase inacetyl-CoA, occurred after the switch. The acetate productionrate decreased throughout the culture with a temporary increaseat the switch point, but the intracellular acetate poolremained relatively constant.
Two mixtures of amino acids were fed together with glucosein fed-batch cultures ofE. coliW3110 pRIT44T2, expressing the recombinantprotein ZZT2. One mixture contained 20 amino acids and theother 5 so-called 'protein amino acids': Ala, Arg, Met, His andPhe. Although the amino aids increased the cell yield anddecreased the proteolysis rate in both cases, ZZT2 productionwas decreased. A decrease of ZZT2 synthesis rate is consideredto be the reason. Further studies of the 5 amino acidsindicated that a few amino acids disturb metabolism.
Carbon mass balances were calculated in glucose limitedfed-batch cultures ofE. coli. In the end, the carbon recovery was ~90% basedon biomass, CO2and acetate, but ~100% if the all carbon in themedium was included. Outer membrane (OM) constituents,lipopolysaccharide, phospholipids, and carbohydratescontributed to 63% of the extracellular carbon. Little celllysis occurred and the unidentified (~30%) carbon was assumedto constitute complex carbohydrates. A novel cultivationtechnique Temperature-Limited Fed-Batch (TLFB) is developed toprevent OM shedding in high-cell density cultures.
Keywords: Escherichia coli, amino acids, glycine, quorumsensing, metabolic switch, metabolite pools, carbon balance,outer membrane, lipopolysaccharide, batch culture, fed-batchculture
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Souza, Jackson Antônio Marcondes de [UNESP]. "Perfil transcricional de Bradyrhizobium elkanii SEMIA 587 in vitro e em simbiose com soja (Glycine max L. Merrill) através de microarranjo de DNA." Universidade Estadual Paulista (UNESP), 2006. http://hdl.handle.net/11449/103907.
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O nitrogênio é o nutriente requerido em maior quantidade para a cultura da soja. Avanços nas pesquisas de melhoramento genético vegetal e microbiologia do solo permitiram expandir o uso de inoculantes comerciais contendo estirpes de Bradyrhizobium japonicum e Bradyrhizobium elkanii. Estas bactérias infectam as raízes da planta e induzem a formação de nódulos, que abrigam a forma bacterióide, diferenciada da bactéria, responsável pela fixação simbiótica do nitrogênio. Informações sobre processos bioquímicos envolvidos no metabolismo da relação simbiótica podem ser adquiridas através de análises globais de expressão gênica. Para esta finalidade, destaca-se a tecnologia de microarranjo de DNA para detecção de genes diferencialmente expressos em larga escala. O objetivo geral deste trabalho foi identificar genes diferencialmente expressos, por meio de microarranjos de DNA, em Bradyrhizobium elkanii SEMIA 587 cultivada em diferentes meios de cultura, RDM (Rhizobia Defined Medium), TY (Triptone-Yeast Medium) e YMB (Yeast-Mannitol Medium), e em bacterióides isolados de nódulos de soja em diferentes períodos de desenvolvimento, 13, 28 e 48 dias após inoculação. Para esta finalidade, a partir do seqüenciamento de DNA genômico de B. elkanii, um microarranjo (Be587) foi gerado contendo 2654 genes. Em meio RDM, a bactéria confrontou-se com a necessidade de se adaptar e sintetizar suas subunidades formadoras de macromoléculas a partir de uma única fonte de carbono, refletindo em um metabolismo mais ativo nas fases lag e log. Por outro lado, em meio TY, as células cultivadas na presença de uma boa fonte de carbono e energia cresceram rapidamente esgotando os recursos disponíveis no meio, 8 o que pode ter causado uma situação de estresse que se refletiu na identificação...
Nitrogen is the most required nutrient by soybean culture. Advanced researches in genetic plant breeding and soil microbiology allowed the expansion in commercial inoculants applications containing strains of Bradyrhizobium japonicum and Bradyrhizobium elkanii. These bacteria infect plant roots and induce nodule formation which home the differentiated bacteria, named bacteroid. The bacteroid in turn is responsible for symbiotic nitrogen fixation. Biochemical knowledge about processes of symbiotic regulation can be acquired by global analysis of gene expression. To achieve such information, the DNA microarray technology, used for detection of differentially expressed genes in large scale, was used. The purpose of this work was identificate differentially expressed genes of Bradyrhizobium elkanii SEMIA 587, grown under different media conditions, such as RDM (Rhizobia Defined Medium), TY (Triptone- Yeast Medium) and YMB (Yeast-Mannitol Medium), and in bacteroids from soybean nodules at different developmental stages, 13, 28 e 49 days after inoculation. For this purpose, the DNA microarray Be587 with 2654 genes was generated from B. elkanii genomic DNA. In RDM medium the bacterium was confronted with the need of adaptation and building of macromolecules subunits from a single carbon source, what was reflected in a more active metabolism in lag and log phases. In turn, in TY medium with good carbon and energy sources the cells grew fastly and exhaust the medium sources available. Such condition can submitted the bacterial cells to a stress condition that reflected in the identification of higher number differentially expressed genes. At different bacteroids stages, the analysis detected genes related to nodulation and 10 nitrogen fixation regulation more than structural genes. Inasmuch, an organic nitrogen recycle might be involved... (Complete abstract, click electronic access below)
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Ticak, Tomislav. "Anoxic quaternary amine utilization by archaea and bacteria through a non-L-pyrrolysine methyltransferase; insights into global ecology, human health, and evolution of anaerobic systems." Miami University / OhioLINK, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=miami1429897518.
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Souza, Jackson Antônio Marcondes de. "Perfil transcricional de Bradyrhizobium elkanii SEMIA 587 "in vitro" e em simbiose com soja (Glycine max L. Merrill) através de microarranjo de DNA /." Jaboticabal : [s.n.], 2006. http://hdl.handle.net/11449/103907.
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Orientador: Eliana Gertrudes de Macedo LemosBanca: Maria José Valarini
Banca: Norma Gouvêa Rumjanek
Banca: Lúcia Maria Carareto Alves
Banca: Fátima Maria de Souza Moreira
Resumo: O nitrogênio é o nutriente requerido em maior quantidade para a cultura da soja. Avanços nas pesquisas de melhoramento genético vegetal e microbiologia do solo permitiram expandir o uso de inoculantes comerciais contendo estirpes de Bradyrhizobium japonicum e Bradyrhizobium elkanii. Estas bactérias infectam as raízes da planta e induzem a formação de nódulos, que abrigam a forma bacterióide, diferenciada da bactéria, responsável pela fixação simbiótica do nitrogênio. Informações sobre processos bioquímicos envolvidos no metabolismo da relação simbiótica podem ser adquiridas através de análises globais de expressão gênica. Para esta finalidade, destaca-se a tecnologia de microarranjo de DNA para detecção de genes diferencialmente expressos em larga escala. O objetivo geral deste trabalho foi identificar genes diferencialmente expressos, por meio de microarranjos de DNA, em Bradyrhizobium elkanii SEMIA 587 cultivada em diferentes meios de cultura, RDM (Rhizobia Defined Medium), TY (Triptone-Yeast Medium) e YMB (Yeast-Mannitol Medium), e em bacterióides isolados de nódulos de soja em diferentes períodos de desenvolvimento, 13, 28 e 48 dias após inoculação. Para esta finalidade, a partir do seqüenciamento de DNA genômico de B. elkanii, um microarranjo (Be587) foi gerado contendo 2654 genes. Em meio RDM, a bactéria confrontou-se com a necessidade de se adaptar e sintetizar suas subunidades formadoras de macromoléculas a partir de uma única fonte de carbono, refletindo em um metabolismo mais ativo nas fases lag e log. Por outro lado, em meio TY, as células cultivadas na presença de uma boa fonte de carbono e energia cresceram rapidamente esgotando os recursos disponíveis no meio, 8 o que pode ter causado uma situação de estresse que se refletiu na identificação... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Nitrogen is the most required nutrient by soybean culture. Advanced researches in genetic plant breeding and soil microbiology allowed the expansion in commercial inoculants applications containing strains of Bradyrhizobium japonicum and Bradyrhizobium elkanii. These bacteria infect plant roots and induce nodule formation which home the differentiated bacteria, named bacteroid. The bacteroid in turn is responsible for symbiotic nitrogen fixation. Biochemical knowledge about processes of symbiotic regulation can be acquired by global analysis of gene expression. To achieve such information, the DNA microarray technology, used for detection of differentially expressed genes in large scale, was used. The purpose of this work was identificate differentially expressed genes of Bradyrhizobium elkanii SEMIA 587, grown under different media conditions, such as RDM (Rhizobia Defined Medium), TY (Triptone- Yeast Medium) and YMB (Yeast-Mannitol Medium), and in bacteroids from soybean nodules at different developmental stages, 13, 28 e 49 days after inoculation. For this purpose, the DNA microarray Be587 with 2654 genes was generated from B. elkanii genomic DNA. In RDM medium the bacterium was confronted with the need of adaptation and building of macromolecules subunits from a single carbon source, what was reflected in a more active metabolism in lag and log phases. In turn, in TY medium with good carbon and energy sources the cells grew fastly and exhaust the medium sources available. Such condition can submitted the bacterial cells to a stress condition that reflected in the identification of higher number differentially expressed genes. At different bacteroids stages, the analysis detected genes related to nodulation and 10 nitrogen fixation regulation more than structural genes. Inasmuch, an organic nitrogen recycle might be involved... (Complete abstract, click electronic access below)
Doutor
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Bružytė, Simona. "Arthrobacter globiformis glicino betaino katabolizmo genų tyrimas." Master's thesis, Lithuanian Academic Libraries Network (LABT), 2009. http://vddb.library.lt/obj/LT-eLABa-0001:E.02~2006~D_20090908_193915-70870.
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Arthrobacter sp. yra dirvoje paplitusios bakterijos, kurios gali prisitaikyti prie nepalankių aplinkos sąlygų, tokių kaip osmosinis šokas ar maisto medžiagų trūkumas. Osmoreguliacijoje daugelis organizmų naudoja glicino betainą. Yra žinoma, kad Arthrobacter globiformis bakterijos gali panaudoti glicino betainą kaip anglies ir azoto šaltinį, tačiau nebuvo aišku, ar Arthrobacter globiformis gali šią medžiagą naudoti kaip osmoprotektorių. Iš Arthrobacter globiformis buvo klonuotas DNR fragmentas, kuriame buvo identifikuoti genai, kurie tiesiogiai ir netiesiogiai dalyvauja glicino betaino katabolizme. Struktūrinė genų organizacija leidžia manyti, kad šios bakterijos gali panaudoti glicino betainą kaip osmoprotektorių, Taip pat buvo įdomu patikrinti, ar tokią funkciją glicino betainas gali vykdyti ir kitose A. globiformis giminingose bakterijose. Tyrimui buvo pasirinkti tipiniai Arthrobacter genties bakterijų kamienai (A. atrocyaneus, A. citreus, A. crystalopoietes, A. globiformis, A. ramosus, A. sulfureus bei Arthrobacter spp. (KA3, P3, KA2V2, PY22, KA2, GAZ21, P2G, KA4, KA2V3, GAZ3, PRH1, PY21, VM22, VP23, RD1, VM22, VP22, VP3, VPW7, VPS4, 96, 94, 85, 68M, 83 68B, BL-3 ir 1-IN). Darbo metu paaiškėjo, kad šį junginį apsaugai nuo osmosinio streso naudoja tik dalis Arthrobacter genties bakterijų. Glicino betainas artrobakterėse dažniau naudojamas kaip anglies šaltinis.Arthrobacter spp. are wide-spread soil bacteria, which are adapted to grow under osmotic stress and shortage in available carbon (energy) sources. Many organisms utilise glycine betaine as an osmoprotectant. It is known, that Arthrobacter globiformis can use glycine betaine as a sole carbon and nitrogen source, however, it is unclear whether this bacterium use this substrate as osmoprotectant. A DNA fragment harbouring genes, which directly and indirectly are involved in glycine betaine degradation, was cloned from A. globiformis. The structural organization of these genes suggested, that this bacterium could use glycine betaine as an osmoprotectant. So, it was tested if other Arthrobacter spp. strains use glycine betaine to protect from osmotic stress. Typical Arthrobacter sp. strains (A. atrocyaneus, A. citreus, A. crystalopoietes, A. globiformis, A. ramosus, A. sulfureus and Arthrobacter spp. (KA3, P3, KA2V2, PY22, KA2, GAZ21, P2G, KA4, KA2V3, GAZ3, PRH1, PY21, VM22, VP23, RD1, VM22, VP22, VP3, VPW7, VPS4, 96, 94, 85, 68M, 83 68B, BL-3 and 1-IN) were chosen in this study. It turned out, that less than a half of studied strains use glycine betaine as osmoprotectant. The arthrobacters used glycine betaine more as a sole carbon source than as an osmoprotectant.
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Fraga, Diego Severo. "Respostas de cultivares de soja ao resíduo da mistura dos herbicidas imazapyr e imazapic." Universidade Federal de Pelotas, 2016. http://repositorio.ufpel.edu.br:8080/handle/prefix/3353.
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A rotação da cultura da soja ao arroz irrigado e sucessão com azevém pode ser considerado opção para manejo de plantas daninhas em área de terras baixas, principalmente o arroz-vermelho. Além disso, o surgimento de novas tecnologias de soja resistente a herbicidas do grupo das imidazolinonas como a Cultivance® pode auxiliar o produtor de sementes no controle de plantas daninhas quando associadas a sistemas de manejo como “plante-aplique” e “aplique-plante”. Dessa forma, os objetivos da pesquisa foram avaliar a fitotoxicidade, os componentes da produtividade e a qualidade fisiológica das sementes de cultivares de soja contendo as tecnologias Cultivance®, tolerância a sulfonilureias e Roundup Ready®, frente ao resíduo de doses crescentes da mistura dos herbicidas imazapyr e imazapic, aplicado no manejo da cultura do arroz irrigado; avaliar a permanência do resíduo do herbicida no solo em virtude do acúmulo de massa da matéria seca da parte aérea de plantas de azevém, semeadas em sucessão com a cultura da soja; quantificar a atividade da enzima ALS em cultivares de soja resistentes ou suscetíveis ao herbicida imazapic; avaliar as alterações nos metabólitos secundários de plantas de soja contendo essas tecnologias, submetidas a mistura dos herbicidas imazapyr e imazapic nos sistemas de manejo “plante-aplique e “aplique-plante”. Para isso, foram realizados experimentos em campo, casa-de-vegetação e laboratório da Universidade Federal de Pelotas, Embrapa Clima Temperado e Embrapa Trigo, utilizando diferentes cultivares de soja submetidas ao herbicida imazapyr+imazapic. Os resultados demonstraram que a cultivar de soja contendo a tecnologia Cultivance® é resistente a mistura dos herbicidas imazapyr e imazapic aplicado no manejo do arroz irrigado, porém, as demais cultivares são suscetíveis. O resíduo do herbicida afeta a produtividade e a qualidade fisiológica da semente das cultivares estudadas. A massa da matéria seca da parte aérea do azevém, é reduzida na presença do resíduo do herbicida. O KM (concentração de piruvato) da enzima ALS das cultivares de soja são semelhantes, contudo o Vmáx (velocidade máxima da reação) da cultivar resistente é menor, comparado às demais, supondo-se que a resistência da cultivar contendo a tecnologia Cultivance® pode ser decorrente da insensibilidade da enzima ALS ao herbicida imazapic. As cultivares suscetíveis apresentam, em geral, maior teor de clorofila e sofrem maior ação das espécies reativas de oxigênio, comparadas a resistente. A elevação da dose da mistura dos herbicidas imazapyr e imazapic aumenta a ação do sistema antioxidante enzimático, por meio das enzimas superóxido dismutase, catalase e ascorbato peroxidase, independente do sistema de manejo.
Soybean crop rotation with irrigated rice and sucession with ryegrass may be considered an option to weed management in flooded area, mainly for red rice. Besides that new technologies appearance of soybean herbicide resistant of imidazolinone group such as Cultivance® may help seed farmers in the weed control when associated to management systems such as “plant-apply” and “apply-plant”. Thus the reseach objectives were evaluate phytotoxicity, yield components and physiological quality of soybean seeds’ cultivars containing Cultivance®, sulfonylurea tolerant and Roundup Ready® technologies, as affected by carryover of increasing doses of mixture of herbicides imazapyr and imazapic spraying in management of irrigated rice crop; evaluate herbicide carryover in soil due ryegrass shoot dry matter, seeded in sucession with soybean crop; quantify ALS enzyme activity in soybean cultivars resistants or susceptibles to imazapic herbicide; evaluate the changes on secondary metabolism of soybean plants containing these technologies, submmited to mixture of herbicides imazapyr and imazapic in “plant-apply” and “apply-plant” management systems. Therefore experiments were performed on field, greenhouse and laboratories in the Federal University of Pelotas, Embrapa Temperate Climate and Embrapa Wheat, using differents soybean cultivars submmited to imazapyr+imazapic herbicide. The results showed that soybean cultivar containing Cultivance® technology is resistant to mixture of herbicides imazapyr and imazapic sprayed in management of irrigated rice, but the others cultivars are susceptibles. The herbicide carryover affect the crop yield and physiological quality of seed of the studied cultivars. Ryegrass shoot dry matter is affected by herbicide carryover. ALS enzyme KM (pyruvate concentration) of soybean cultivars are similar, however the Vmáx (maximum reaction speed) of resistant cultivar is minor, compared with the other cultivars, supposing that the resistance of the cultivar containing Cultivance® technology may be due insensibility ALS enzyme to imazapic herbicide. The susceptibles cultivars show, in general, more chlorophyll and suffer more action of reactives oxygen species, compared to resistant. The increasing dose of mixture of herbicides imazapyr and imazapic enhance the action of enzymatic antioxidant system, by means of superoxide dismutase, catalase and ascorbate peroxidase enzymes, independent of management system.
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Borella, Junior. "Adaptações metabólicas de genótipos de soja em resposta à deficiência de oxigênio e envolvimento do nitrato." Universidade Federal de Pelotas, 2015. http://repositorio.ufpel.edu.br:8080/handle/prefix/3589.
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES
O alagamento é um fator ambiental comum que causa deficiência de oxigênio às plantas, levando a uma inibição da respiração e redução do status energético celular, desencadeando uma série de mudanças no metabolismo do carbono e do nitrogênio. Além disso, alterações no fluxo de elétrons da cadeia de transporte de elétrons mitocondrial e cloroplastídica levam a produção de espécies reativas de oxigênio (EROs) que podem ocasionar vários danos ao metabolismo celular. No entanto, a aplicação exógena de nitrato tem sido reportada por promover efeitos benéficos em muitas espécies de plantas sob condições de hipóxia. Embora muitos estudos tenham sido envidados com soja, pouco se sabe a respeito das alterações metabólicas primárias do carbono e do nitrogênio que permitem diferenciar genótipos contrastantes ao alagamento e os efeitos no sistema antioxidante pela aplicação exógena de nitrato nas plantas. Considerando o exposto, os objetivos deste trabalho foram: I – avaliar mudanças no metabolismo do carbono e do nitrogênio e sua relação com a enzima alanina aminotransferase (AlaAT) em genótipos de soja nodulada; II – verificar possíveis efeitos benéficos no metabolismo antioxidante em plantas cultivadas na presença de nitrato (plantas não-noduladas) e na ausência de nitrato (plantas noduladas). Para isso, dois experimentos foram conduzidos em casa de vegetação com plantas de soja (Glycine max (L.) Merril) sob condições naturais de luz e temperatura. Experimento I: plantas noduladas de soja, nutridas na ausência de N mineral (Fundacep 53 RR – tolerante e BRS Macota – sensível) foram cultivadas em vermiculita e transferidas para sistema hidropônico, no estádio reprodutivo R2. O sistema radicular das plantas foi submetido à hipóxia pelo borbulhamento de nitrogênio gasoso na solução nutritiva diluída 1/3 da concentração normal, por 24 e 72 h. Para recuperação, após 72 h de hipóxia as plantas retornaram para vermiculita por 24 e 72 h. Foram avaliados, em raízes e nódulos, metabólitos fermentativos e ácidos orgânicos (GC-MS), aminoácidos (HPLC), expressão relativa dos genes (RT-PCR) e atividade da enzima AlaAT. Fundacep 53 RR acumulou mais teores de piruvato e lactato que BRS Macota e embora a composição de aminoácidos não tenha diferido entre os genótipos, foi observado uma ligação entre a glicólise e o ciclo dos ácidos tricarboxílicos via indução dos genes e atividade da AlaAT, que, posteriormente, levou ao acúmulo de succinato em raízes de Fundacep 53 RR, podendo aumentar o ganho energético em relação à BRS Macota sob hipóxia. Experimento II: A condução experimental adotada foi semelhante ao experimento I, no entanto conduzido com plantas noduladas e não-noduladas (nutridas com nitrato) de soja, de ambos os genótipos. Foi avaliado o sistema antioxidante em raízes e folhas através da atividade das enzimas superóxido dismutase (SOD), ascorbato peroxidase (APX), catalase (CAT), glutationa redutase (GR), guaiacol peroxidase (GPOD) e glutationa S-transferase (GST), o conteúdo de ascorbato reduzido e ascorbato total, bem como conteúdo de superóxido (O2), peróxido de hidrogênio (H2O2) e peroxidação de lipídeos. O sistema antioxidante foi fortemente induzido nas raízes das plantas nutridas com nitrato de ambos os genótipos, com elevada atividade de SOD, APX, CAT, GR e GPOD, bem como o aumento do conteúdo de ascorbado reduzido e total e diminuição da produção de EROs em condições de hipóxia e de recuperação, enquanto que nas folhas de plantas noduladas e não-noduladas foi observado um ligeiro aumento nos componentes enzimáticos e não enzimáticos antioxidantes. O nitrato exerce efeitos benéficos em plantas de soja em condições de hipóxia e consequentemente na recuperação por induzir o sistema antioxidante nas raízes, permitindo modular os possíveis danos oxidativos causados pela produção de EROs, além de poder prolongar a tolerância dessas plantas.
Waterlogging is a common environmental stress which causes oxygen deprivation in plants leading to an inhibition of the mitochondrial respiration. It leads to a reduction of cellular energy status triggering changes at different levels of carbon and nitrogen metabolism. In addition, it leads to electron scape from the mitochondrial and chloroplast electron transport chain, producing reactive oxygen species (ROS) which cause severe oxidative damage to cells. However, exogenous nitrate supply has been reported to promoting beneficial effects in several plant species under hypoxic conditions. Although many studies have been carried out with soybean, a little is known about the primary metabolic changes in carbon and nitrogen metabolism, which may differ between tolerant and sensitive plant genotypes in response to waterlogging and the effects on antioxidant system in nitrate-supplied plants in comparison to non-nitrate-supplied plants. Thus, the aims of this study were: I – to evaluate the hypoxia-induced alterations of carbon and nitrogen metabolism and its relation with alanine aminotransferase (AlaAT) enzyme in nodulated soybean genotypes; II – to verify possible beneficial effects on antioxidant metabolism in nitrate-supplied plants (non-nodulated plants) in comparison to plants growing in absence of nitrate (nodulated plants). For that, two experiments were carried out in greenhouse under natural light and temperature conditions. Experiment I: Nodulated soybean plants (Fundacep 53 RR – tolerant and BRS Macota – sensitive) were grown in vermiculite and transferred to hydroponic system at reproductive stage. Root system was subjected to hypoxia by flushing N2 gas into the solution for 24 or 72 h. For the recovery, after 72 h in hypoxia, plants returned to normoxic conditions by transferring back to vermiculite for 24 and 72 h. Root and nodule organic acids and amino acids were analysed by gas chromatography-mass spectrometry and high-performance liquid chromatography, respectively. Relative expression of AlaAT (qRT-PCR) and AlaAT activity were also verified in both genotypes. Plants of Fundacep 53 RR and BRS Macota genotypes responded distinctly upon hypoxia. Fundacep 53 RR presented higher pyruvate and lactate accumulation than BRS Macota, which is indicative of higher glycolytic and fermentation rates in root tissues. Furthermore, Fundacep 53 RR responds more effectively to the recovery by restoring pre-hypoxic levels of the metabolites. Although the amino acid composition did not differ between the genotypes, there was a clear link between glycolysis and the TCA via increase of gene expression and activity of AlaAT enzyme by leading a succinate accumulation in Fundacep 53 RR, wich represents a metabolic advantage compared to BRS Macota under hypoxic stress. Experiment II: was carried out in a similar way of Experiment I, however with plants growing in presence (non-nodulated) and absence (nodulated) of nitrate, for both soybean genotypes. Superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR), guayacol peroxidase (GPOD) and glutathione S-transferase (GST) enzymes; reduced ascorbate and ascorbate redox state; superoxide content (O2•-), hydrogen peroxide (H2O2) and lipid peroxidation were analysed in roots and leaves of both soybean genotypes. Antioxidative system was strongly induced in roots of nitrate-supplied plants of both genotypes, with high activity of superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT), glutathione reductase (GR) and guayacol peroxidase (GPOD), as well as increased ascorbate reduced and ascorbate redox state and decreased ROS production under hypoxia and recovery, while in leaves of nodulated and non-nodulated plants a slight increase on antioxidant system was observed. Furthermore, the results did not show tolerance differences between the genotypes. Nitrate exerts beneficial effects in soybean plants under hypoxic conditions and consequent recovery by inducing the antioxidant system manly in roots, to cope possible oxidative damage caused by ROS production and also can postpone the effects of hypoxia in both genotypes.
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Perez, Gerardo Gomez. "Creatine: Physiology and performance: The health effects of creatine in exercise and human performance." CSUSB ScholarWorks, 2004. https://scholarworks.lib.csusb.edu/etd-project/2453.
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The purpose of this project is to review literature on creatine monohydrate (simply known as creatine/Cr) supplementation and its effects on exercise, human performance, and health. Included in this project is basic information relating to the biochemical and physiological effects of Cr, including possible side effects.
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Srinivasan, Asha R. "Studies into sulfur amino acid and bile salt metabolism in pancreatic and liver diseases : profiles of sulfur amino acids and glutathione in acute pancreatitis : method development for total and oxidized glutathione by liquid chromatography : bile salt profiles in liver disease by liquid chromatography-mass spectrometry." Thesis, University of Bradford, 2010. http://hdl.handle.net/10454/4434.
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Sulfur amino acids have critical function as intracellular redox buffers and maintain homeostasis in the external milieu by combating oxidative stress. Synthesis of glutathione (GSH) is regulated at a substrate level by cysteine, which is synthesized by homocysteine via the transsulfuration pathway. Oxidative stress and diminished glutathione pools play a sustained role in the pathogenesis of acute pancreatitis. One of the aims of this study was to experimentally address the temporal relationship between plasma sulfur amino acid levels in patients suffering from acute pancreatitis. The data indicated low concentration of cysteine initially, at levels similar to those of healthy controls. Glutathione was found reduced whilst cysteinyl-glycine and γ- glutamyl transpeptidase activity were increased in both mild and severe attacks. As the disease progressed, glutathione and cysteinyl-glycine were further increased in mild attacks and cysteine levels correlated with homocysteine and γ-glutamyl transpeptidase activity. The progress of severe attacks was associated with glutathione depletion, reduced γ-glutamyl transpeptidase activity and increased cysteinyl-glycine, that correlated with glutathione depletion. The corollary that ample supply of cysteine and cysteinly-glycine does not contribute towards glutathione synthesis in acute pancreatitis poses an important issue that merits resolution. Heightened oxidative stress and depletion of glutathione rationalized the progression of disease in severe attacks. An upsurge that reactive oxygen species can shift redox state of cells is determined by the ratio of the abundant redox couples reduced and oxidized glutathione (GSH: GSSG) in cell. The study reported a novel methodology for quantification of total oxidized glutathione (tGSSG) and total glutathione (tGSH) in whole blood using reverse phase high performance liquid chromatography. The novelty of the method is ascertained by the use of a mercaptan scavenger 1, methyl-2-vinyl-pyridinium trifluromethanesulfonate for the total oxidized glutathione determination. The results reported permit quantitation of tGSSG and tGSH and was applied to a control group. Finally, the study was also focussed in developing a liquid chromatography-mass spectrometric method to evaluate free and conjugated bile acids in patients suffering from various degrees of cholestatic-hepatobiliary disorders. The study reported low levels of ursodeoxycholic acid (UDCA) and slightly high levels of lithocholic acid (LCA). All the primary bile acids seem to be conjugated with glycine and taurine amino acid.
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Menegassi, Angela. "Identificação, caracterização estrutural e funcional de aquaporinas de soja (Glycine max) e seu envolvimento no metabolismo de ureia." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2015. http://hdl.handle.net/10183/131970.
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Aquaporinas, também conhecidas como Major Intrinsic Proteins (MIPs), são proteínas de membrana distribuídas em todos os domínios da vida, responsáveis pelo transporte de água e pequenos solutos. Em plantas superiores, essa família é particularmente diversa e pode ser classificada em cinco subfamílias: proteínas intrínsecas de membrana plasmática, proteínas intrínsecas de tonoplasto, proteínas intrínsecas do tipo nodulina-26, proteínas intrínsecas pequenas e proteínas intrínsecas X. Esses canais apresentam uma estrutura bastante conservada, exibindo, no centro do poro, duas regiões importantes para a seletividade: os motivos NPA e o filtro seletivo. No presente trabalho, sessenta e seis genes de aquaporinas foram identificados no genoma da soja e classificados por homologia de sequência nas cinco subfamílias estabelecidas. As estruturas tridimensionais das MIPs de soja foram determinadas por modelagem comparativa e o raio dos poros e os resíduos importantes para a especificidade foram identificados. Ensaios funcionais utilizando expressão heteróloga em Saccharomyces cerevisiae foram realizados para quatro aquaporinas, GmTIP1;9, GmTIP2;5, GmTIP3;2 e GmNIP2;2, demonstrando que esses genes codificam canais funcionais capazes de transportar água, peróxido de hidrogênio e ureia. A ureia possui um papel muito importante na agricultura por ser o fertilizante nitrogenado mais utilizado mundialmente. A identificação de transportadores de ureia em plantas ainda é recente e, além de um transportador específico, somente as aquaporinas estão relacionadas ao transporte de ureia em plantas. Apesar disso, a relevância fisiológica desse transporte pelas MIPs é ainda motivo de debate. Empregando uma abordagem baseada em PCR quantitativo, foi demonstrado que os níveis de expressão de GmTIP1;9, GmTIP3;2 e GmNIP2;2, mas não de GmTIP2;5, são alterados em raízes de soja cultivada em solução nutritiva contendo ureia como a única fonte de nitrogênio. Além disso, os níveis dos transcritos de GmNIP2;2 são aumentados na parte aérea das plantas, indicando o possível envolvimento dessa aquaporina na redistribuição de ureia nos tecidos. Outros genes de aquaporinas de soja também apresentaram maior expressão nessas condições. Esses resultados sugerem o envolvimento de aquaporinas no metabolismo de ureia em plantas e apontam essas proteínas como potenciais alvos para manipulação genética visando o aumento da eficiência do uso de nitrogênio em sistemas adubados com ureia e a melhoria da produção agrícola.Aquaporins, also known as Major Intrinsic Proteins (MIPs), are membrane proteins distributed in all kingdoms of life, responsible for water and small solutes transport. In higher plants, this family is particularly diverse and can be classified into five subfamilies: plasma membrane intrinsic proteins (PIPs), tonoplast intrinsic proteins (TIPs), nodulin-26-like intrinsic proteins (NIPs), small basic intrinsic proteins (SIPs) and X intrinsic proteins (XIPs). These channels present a highly conserved structure, displaying, in the center of the pore, two regions important for selectivity: the NPA motifs and the selective filter. In the present work, sixty-six aquaporin genes were identified in soybean genome and classified by sequence homology into the five established subfamilies. The three-dimensional structure of soybean MIPs was determined by homology modeling and the pore radius and residues influencing substrate specificity were identified. Functional assays using heterologous expression in Saccharomyces cerevisiae were performed for four aquaporins, GmTIP1;9, GmTIP2;5, GmTIP3;2 and GmNIP2;2, indicating that these genes codify functional channels capable of transporting water, hydrogen peroxide and urea. Urea plays an important role in agriculture because it is the most used nitrogen fertilizer worldwide. Plant urea transporters are just beginning to be identified and, to date, besides a specific transporter, only aquaporins are linked to urea transport in plants. Nevertheless, the physiological relevance of this transport is still in debate. Using a quantitative PCR approach, we have shown that the expression levels of three urea transporting MIPs, GmTIP1;9, GmTIP3;2 and GmNIP2;2, but not GmTIP2;5, are altered in soybean roots when urea is the sole nitrogen source. Moreover, transcript levels of GmNIP2;2 were also up-regulated in shoots, indicating the possible involvement of this aquaporin in urea redistribution to other plant tissues. Several other soybean aquaporin genes were up-regulated in this condition as well. Taken together, these results suggest the involvement of aquaporins in urea metabolism in plants and place these proteins as potential targets for plant manipulation to improve urea based nitrogen use efficiency and crop production.
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Amarante, Luciano do. "Transporte e metabolismo de glutamina em soja [Glycine max (L.) Merril] : uma relação estreita com a fixação simbiotica do nitrogenio atmosferico." [s.n.], 2002. http://repositorio.unicamp.br/jspui/handle/REPOSIP/315244.
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Orientador : Ladaslav SodekTese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: Plantas de soja noduladas fixando plenamente o N2, cultivadas na ausência de N-combinado, contêm níveis relativamente altos de GLN como forma de transporte de N no xilema (cerca de 20-25 em mol % do total de aminoácidos) comparadas a plantas não noduladas nutridas com NO3- (cerca de 1-2 em mol % do total de aminoácidos). Experimentos que levaram à redução da fixação de N2, avaliados pela redução do teor de URE na seiva do xilema, como a aplicação de NO3 -, transferência das plantas para sistema hidropônico com aeração, inundação do sistema radicular, substituição do ar da rizosfera com N2 ou mistura argônio:oxigênio (80:20), conduziram a uma redução severa e específica dos níveis de GLN. A remoção manual dos nódulos do sistema radicular produziu o mesmo efeito. Além disso, durante o crescimento das plantas foi observado um aumento substancial dos níveis de GLN correspondentes à fase de formação dos nódulos e o início da fixação plena de N2 em soja, em paralelo ao aumento de ureídeos e atividade da GS. Aumentos concomitantes dos níveis de GLN e de URE também foram observados quando plantas noduladas de soja, cultivadas em vermiculita sem N e transferidas para hidroponia, na presença ou ausência de N, ou inundadas, foram replantadas em vermiculita ou tiveram a inundação suprimida através da drenagem. A NADH-GOGAT nos nódulos parece ter uma relação direta com níveis de GLN transportados no xilema, partindo da observação da queda e posterior elevação da atividade da enzima em paralelo aos níveis de GLN nos tratamentos de hidroponia na presença e ausência de NO3 ¿ e posterior replantio em vermiculita sem N, respectivamente. As mudanças de atividade da GS em nódulos foram muito menos pronunciadas, sugerindo elevada estabilidade da enzima sob os tratamentos com hidroponia. A composição nitrogenada da seiva do xilema das plantas inundadas de soja mostrou uma queda considerável dos níveis de GLN, 10 minutos após início do tratamento, alcançando valores muito baixos dentro de 30 min. Uma inibição concomitante do teor de URE foi obtida nesses tratamentos. Apesar da evidência de que a atividade dos nódulos esteja relacionada às flutuações dos níveis de GLN, foram encontrados teores mínimos desse aminoácido nos nódulos, não sofrendo variações detectáveis com a aplicação dos tratamentos. As reduções de GLN no xilema também não mostraram uma correspondência com os níveis de GLN do floema, raízes e folhas. Os tratamentos inibitórios à fixação de N2 como a aplicação de NO3 - e inundação também causaram uma redução dos níveis de GLN em espécies de leguminosas exportadoras de amidas e em ambos os teores de GLN e de URE em outras espécies de leguminosas que exportam preferencialmente URE, sugerindo ser este um efeito comum em leguminosas que transportam GLN. Coletivamente, as mudanças da composição aminoacídica do xilema frente aos diferentes tratamentos utilizados indicam que o nível de GLN transportado no xilema está fortemente associado com a atividade dos nódulos
Abstract: Nodulated soybean plants, fully dependent on N2 fixation, cultivated with N deficient nutrient solution, contain relatively high levels of GLN in N transport of the xylem (around 20-25 mol % of total amino acids) compared to non-nodulated plants grown on nitrate (around 1-2 mol % of total amino acids). Experiments that lead to reduced N2 fixation, judged by xylem sap ureide levels, such as NO3 - application, transfer of plants to aerated hydroponic culture, waterlogging of the root system, substitution of air in the rhizosphere with nitrogen or a mixture of argon:oxygen (80:20), lead to severe and specific reduced GLN levels. Manual removal of the nodules from the root system produced the same effect. Furthermore, during the growth cycle of the plants, a substantial increase in GLN levels was observed corresponding to the phase of nodule formation and beggining of N2 fixation, in parallel with increases in ureide levels and GS activity. Concomitant increases in GLN and ureide levels were observed, when nodulated soybean plants cultivated in vermiculite without N and transfered to hydroponic culture with or without N or waterlogging, were returned to vermiculite or the waterlogging treatment was suppressed by drainage. In nodules, NADH-GOGAT activity seems to be directly correlated with GLN levels transported in the xylem, judging by the decline and posterior elevation of enzyme activity in parallel with GLN levels in the hydroponic treatments with or without N and when plants were returned to vermiculite without N, respectively. Changes in GS activity in nodules were much less pronounced suggesting high enzyme stability under hydroponic treatment. The xylem sap nitrogen composition of waterlogged plants showed a considerable fall in glutamine levels, ten minutes after the treatment had initiated, reaching very low levels within 30 minutes. A concomitant inhibition in ureide contents was observed. Despite the evidence that nodule activity underlies the flutuations of GLN levels in the xylem, very low levels in the nodule were found, which suffered no detectable variation after the treatments applied. Decreases in xylem GLN were not associated with GLN levels in phloem, roots or leaves. Inhibitory effects on nitrogen fixation such as NO3 - application and waterlogging caused a reduction in GLN in amide exporting legumes and in both GLN and ureide levels in other URE exporting legume species, suggesting a common effect in GLN exporting legumes. Collectively, changes in amino acid xylem composition under the several treatments tested indicated that GLN levels transported in xylem are largely associated with nodule activity
Doutorado
Doutor em Biologia Vegetal
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Fassini, Priscila Giácomo [UNESP]. "Estudo experimental do efeito da proteína glicinina da soja (Glycine max L.) no metabolismo do colesterol." Universidade Estadual Paulista (UNESP), 2010. http://hdl.handle.net/11449/88658.
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fassini_pg_me_arafcf.pdf: 613112 bytes, checksum: f00daa1b71f5ee79d5673101f5d6e0e8 (MD5)Universidade Estadual Paulista (UNESP)
A globulina 11S (glicinina) é a proteína predominante da soja e tem sido extensivamente caracterizada. Além das suas propriedades nutricionais, há evidências de sua participação na redução das concentrações lipídicas do soro. Por outro lado, o fármaco rosuvastatina tem suas ações demonstradas no metabolismo lipídico, por reduzir a fração LDL-colesterol, bloqueando a chave do processo de produção de colesterol hepático. Embora a eficiência do medicamento seja comprovada, é importante considerar a alimentação como uma possível ação somatória, tendo em vista os graves problemas ocasionados pelas alterações lipídicas no organismo. À vista do exposto, a presente pesquisa teve por objetivo estudar o efeito da glicinina isolada da soja em comparação ao fármaco rosuvastatina em animais submetidos à dieta hipercolesterolêmica. Foram elaboradas duas dietas, uma padrão, contendo caseína como proteína (AIN-93M), e outra hipercolesterolêmica, composta pela dieta padrão adicionada de 1% de colesterol e 0.5% de ácido cólico. A proteína glicinina de soja (300 mg/kg de peso corporal) e o fármaco rosuvastatina (10 mg/Kg de peso corporal), ambos solubilizados em solução salina, foram administrados por gavagem. Foram utilizados ratos machos Wistar mantidos em gaiolas individuais sob condições adequadas. Os animais foram separados em cinco grupos (n = 9): 1) grupo padrão (STD) recebeu a dieta padrão; 2) grupo hipercolesterolêmico (HC) recebeu apenas a dieta hipercolesterolêmica; 3) grupo (HC+11S) recebeu a dieta e a proteína glicinina da soja; 4) grupo (HC+ROS) recebeu a dieta e o fármaco; 5) grupo (HC+ROS+11S) recebeu a dieta, a proteína e o fármaco. Ao final de 28 dias os animais foram sacrificados e o sangue removido para análises bioquímicas de colesterol total (CT), HDL-colesterol (HDL-C) e triglicérides (TG) plasmático, CT e TG hepático...
The 11S globulin (glycinin) is the predominant protein of soybeans and has been extensively characterized. In addition to its nutritional properties there is evidence of its action in reducing serum lipid concentrations. Moreover, the drug rosuvastatin has demonstrated its effects on lipid metabolism, by reducing LDL cholesterol and blocking the key enzyme in the process of cholesterol production in the liver. Although the efficiency of the drug is proven, it is important to consider the sum of its action and that of food, in view of the serious problems caused by changes in body fat. In view of the foregoing, the aim of the present study was to investigate the effects of isolated soy glycinin, as compared to the drug rosuvastatin, in rats subjected to a hypercholesterolemic diet. Two diets were prepared, a standard diet, containing casein as protein (AIN-93M), and a hypercholesterolemic, consisting of the standard diet plus 1% cholesterol and 0.5% cholic acid. The glycinin (300 mg/kg body weight) and the rosuvastatin (10 mg/kg body weight), both dissolved in saline, were administered by gavage. Male Wistar rats were kept in individual cages under appropriate conditions. The animals were divided into five groups (n=9): 1) standard group (STD) received the standard diet; 2) hypercholesterolemic group (HC) received the hypercholesterolemic diet alone; 3) group HC+11S received the HC diet and the glycinin; 4) group HC+ROS received the HC diet and the drug; 5) group HC+ROS+11S received the HC diet, the protein and the drug. After 28 days, the animals were sacrificed and the blood removed for biochemical analysis of total plasma cholesterol (TC), HDL-cholesterol (HDL-C) and triglycerides (TG), and hepatic TC and TG. The results indicated that the experimental HC diet was able to induce hypercholesterolemia and that a single daily dose of the isolated protein was appropriate for a comparative... (Complete abstract click electronic access below)
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Queiroz, Helena Muller. "Efeito do sal no crescimento e metabolismo de plantas de Glycine max L., cv. IAC 17." [s.n.], 2007. http://repositorio.unicamp.br/jspui/handle/REPOSIP/314922.
Full textAbstract:
Orientador: Claudia Regina Baptista HaddadDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: A salinidade inibe o crescimento das plantas e interfere na produtividade de espécies cultivadas. Com o objetivo de verificar alterações no crescimento e metabolismo, plantas de soja, Glycine max (L) Merr., cv. IAC 17, foram cultivadas na presença de NaCI, nas concentrações de 50 mM, 100 mM e 200 mM. A análise dos parâmetros de crescimento revelou que as raízes de plantas de soja são menos afetadas pelo sal do que a parte aérea. O comprimento e a massa fresca das raízes não foram afetados em concentrações elevadas de sal. Dados na literatura relatam que a manutenção do crescimento normal das raízes sob condições adversas de cultivo demonstra que a planta pode apresentar tolerância ao sal. Sob salinidade houve aumento do conteúdo de água das raízes, o que pode estar relacionado com a maior tolerância ao estresse salino, pois, desta forma, a planta conseguiria diminuir a concentração de sais no citoplasma. Observou-se um decréscimo gradativo na área foliar, massas secas e secas livres de cinzas de folhas, caules e raízes com o aumento da salinidade. O teor de proteínas em folhas de plantas de soja não foi alterado significativamente pela salinidade, em relação ao controle, uma explicação para o fato é a possibilidade de ter havido aumento na síntese de proteínas específicas de estresse. Outra possibilidade é que a duração do experimento não tenha sido suficiente para que fossem observadas alterações na concentração de proteínas. Não houve uma relação clara entre a concentração de açúcares solúveis totais nas raízes e a concentração de NaCl no meio de cultivo, indicando que os açúcares solúveis não devem estar envolvidos no ajustamento osmótico em plantas de soja. Houve redução nas concentrações de nitrato e aminoácidos solúveis totais nas raízes, quando as plantas foram submetidas à salinidade. A redução na concentração de aminoácidos livres totais pode estar ligada à diminuição da absorção de nitrato pelas raízes, devido a uma possível interferência exercida pelo Cl- sobre transportadores de membrana, que limitaria a entrada do nitrato nas raízes. Diante de uma menor absorção de nitrato, haveria limitação na assimilação do nitrogênio necessário para a síntese de aminoácidos. A salinidade provocou alterações no perfil de aminoácidos transportados no xilema. Houve um aumento na concentração de SER, ALA, GABA e PRO e redução de ASN no exsudato do xilema de plantas submetidas aos tratamentos com solução salina. A diminuição da concentração de ASN pode ter ocorrido devido à interconversão deste aminoácido em ALA. Nas raízes de plantas controle, os aminoácidos que apresentaram a maior proporção foram ASN, GABA e GLU, respectivamente e, após a adição de sal ao meio, apenas a concentração do aminoácido ASN diminuiu. A PRO e o GABA têm um possível papel na redução da acidificação do citosol, sob estresse salino. Apesar da elevação observada dos mesmos não foi suficiente para evitar a redução do crescimento sob salinidade, uma vez que houve diminuição da massa seca das plantas de soja
Abstract: Salinity inhibits plant growth and crop production. With the objective of verifying changes in metabolism and growth, soybean plants (Glycine max (L) Merr., cv. IAC 17) were cultivated in the presence of different NaCl concentrations (50 mM, 100 mM and 200 mM). The analysis of growth parameters showed that the roots of soybean plants are less affected by salt than the shoot. The length and fresh matter of roots were not significantly reduced by the increase in salt concentration. Data from the literature indicate that normal growth of roots under adverse conditions may demonstrate the capacity of plants to tolerate salt stress. The increase of water content in roots of soybean plants cultivated at higher salt concentrations can be related with salt tolerance, since the increase of water reduces salt concentration inside the cytoplasm. A gradual decrease was observed in leaf area, dry matter and ash-free dry matter of leaves, stem and roots, with the increase of salinity. The protein concentration of soybean leaves was not significantly changed by the salinity. The explanation for this is probably related to the increase of specific proteins synthesized by the plants under stress conditions. Another possibility is that the experimental period was not enough to allow observations in changes of protein concentrations. There was no clear relationship between total soluble sugars located in the root system and the NaCl concentration, which may indicate that these were not used by the plant for osmotic adjustment. A reduction of nitrate and total soluble amino acids was observed in plants affected by salinity. The reduction of free total amino acids may be related to the decrease of nitrate absorption by the roots, possibly resulting from an interference of Cl- on membrane nitrate transporters. Salinity changed the amino acid profile transported in the xylem. There was an increase in concentration of SER, ALA, GABA and PRO and reduction of ASN in xylem sap of plants under salt stress. The decrease of ASN can be explained by the interconversion of this into ALA. In the roots of control plants the most abundant amino acids were ASN, GABA and GLU, respectively, and after the addition of salt, only the ASN level was reduced. Some authors consider that PRO and GABA have a role in the reduction of cytosol acidification under salinity stress. Although an increase in PRO and GABA were observed, it did not avoid the reduction of growth, as shown by the reduction of dry matter in plants under salt stress
Mestrado
Mestre em Biologia Vegetal
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Fassini, Priscila Giácomo. "Estudo experimental do efeito da proteína glicinina da soja (Glycine max L.) no metabolismo do colesterol /." Araraquara : [s.n.], 2010. http://hdl.handle.net/11449/88658.
Full textAbstract:
Orientador: Aureluce DemonteBanca: Valdir Augusto Neves
Banca: Olga Luisa Tavano
Resumo: A globulina 11S (glicinina) é a proteína predominante da soja e tem sido extensivamente caracterizada. Além das suas propriedades nutricionais, há evidências de sua participação na redução das concentrações lipídicas do soro. Por outro lado, o fármaco rosuvastatina tem suas ações demonstradas no metabolismo lipídico, por reduzir a fração LDL-colesterol, bloqueando a chave do processo de produção de colesterol hepático. Embora a eficiência do medicamento seja comprovada, é importante considerar a alimentação como uma possível ação somatória, tendo em vista os graves problemas ocasionados pelas alterações lipídicas no organismo. À vista do exposto, a presente pesquisa teve por objetivo estudar o efeito da glicinina isolada da soja em comparação ao fármaco rosuvastatina em animais submetidos à dieta hipercolesterolêmica. Foram elaboradas duas dietas, uma padrão, contendo caseína como proteína (AIN-93M), e outra hipercolesterolêmica, composta pela dieta padrão adicionada de 1% de colesterol e 0.5% de ácido cólico. A proteína glicinina de soja (300 mg/kg de peso corporal) e o fármaco rosuvastatina (10 mg/Kg de peso corporal), ambos solubilizados em solução salina, foram administrados por gavagem. Foram utilizados ratos machos Wistar mantidos em gaiolas individuais sob condições adequadas. Os animais foram separados em cinco grupos (n = 9): 1) grupo padrão (STD) recebeu a dieta padrão; 2) grupo hipercolesterolêmico (HC) recebeu apenas a dieta hipercolesterolêmica; 3) grupo (HC+11S) recebeu a dieta e a proteína glicinina da soja; 4) grupo (HC+ROS) recebeu a dieta e o fármaco; 5) grupo (HC+ROS+11S) recebeu a dieta, a proteína e o fármaco. Ao final de 28 dias os animais foram sacrificados e o sangue removido para análises bioquímicas de colesterol total (CT), HDL-colesterol (HDL-C) e triglicérides (TG) plasmático, CT e TG hepático... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The 11S globulin (glycinin) is the predominant protein of soybeans and has been extensively characterized. In addition to its nutritional properties there is evidence of its action in reducing serum lipid concentrations. Moreover, the drug rosuvastatin has demonstrated its effects on lipid metabolism, by reducing LDL cholesterol and blocking the key enzyme in the process of cholesterol production in the liver. Although the efficiency of the drug is proven, it is important to consider the sum of its action and that of food, in view of the serious problems caused by changes in body fat. In view of the foregoing, the aim of the present study was to investigate the effects of isolated soy glycinin, as compared to the drug rosuvastatin, in rats subjected to a hypercholesterolemic diet. Two diets were prepared, a standard diet, containing casein as protein (AIN-93M), and a hypercholesterolemic, consisting of the standard diet plus 1% cholesterol and 0.5% cholic acid. The glycinin (300 mg/kg body weight) and the rosuvastatin (10 mg/kg body weight), both dissolved in saline, were administered by gavage. Male Wistar rats were kept in individual cages under appropriate conditions. The animals were divided into five groups (n=9): 1) standard group (STD) received the standard diet; 2) hypercholesterolemic group (HC) received the hypercholesterolemic diet alone; 3) group HC+11S received the HC diet and the glycinin; 4) group HC+ROS received the HC diet and the drug; 5) group HC+ROS+11S received the HC diet, the protein and the drug. After 28 days, the animals were sacrificed and the blood removed for biochemical analysis of total plasma cholesterol (TC), HDL-cholesterol (HDL-C) and triglycerides (TG), and hepatic TC and TG. The results indicated that the experimental HC diet was able to induce hypercholesterolemia and that a single daily dose of the isolated protein was appropriate for a comparative... (Complete abstract click electronic access below)
Mestre
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Pfrimer, Karina. "Avaliação do metabolismo protéico em idosos brasileiros independentes utilizando a glicina marcada com 15N." Universidade de São Paulo, 2006. http://www.teses.usp.br/teses/disponiveis/17/17138/tde-17082006-153645/.
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O metabolismo protéico em idosos, analisado pela medida da velocidade de reciclagem, é um importante fator para a análise da manutenção da massa muscular e das atividades de vida diária. Dados coletados em idosos apontam uma redução da síntese protéica com o envelhecimento. Outros relatam ser esta mantida e a degradação aumentada. Esta investigação teve por objetivo avaliar o metabolismo protéico de idosos saudáveis e independentes utilizando a glicina, marcada com o isótopo 15N. Sete idosos saudáveis foram estudados.Foram feitas avaliações clínica, nutricional e bioquímica em todos os voluntários, sendo excluídos aqueles portadores de doenças ou usuários de medicamentos que interferissem no metabolismo protéico. Foi oferecida uma dose oral de 200 mg de 15N-Glicina e coletadas amostras de sangue e urina (basal, antes do consumo da glicina, quatro horas e meia e nove horas após a ingestão da glicina). Foram quantificados amônia, uréia e nitrogênio total e as amostras analisadas por espectrometria de massa, para a determinação do enriquecimento isotópico (15N). Os voluntários tinham 65,4 ± 2,8 anos (média ± desvio padrão), quatro mulheres e três homens, com IMC de 22,73 ± 2,4 Kg/m2. Total de nitrogênio excretado de 3,31 ± 0,7 gN/9horas e a ingestão de 7,76 ± 1,0 gN/9horas, o fluxo de nitrogênio 15 foi de 30,36 ± 6,3 gN/9horas, o balanço nitrogenado de 4,46 ± 1,0 g/N. Os valores encontrados nesta pesquisa foram similares aos da literatura para idosos e menores que os referidos para jovens. Este estudo estabeleceu os valores do metabolismo protéico em idosos saudáveis, ingerindo alimentação típica (arroz e carne moída), o que permitirá posteriores estudos de intervenção.Protein metabolism in the elderly, analyzed though the protein turnover rate, is of great importance to muscle trophysm and maintenance of activities of daily living. Some studies in the elderly have shown a reduction of protein synthesis with aging, while others found that it is maintained and degradation increased. Methods using stable isotopes are of great relevance in the research of protein metabolism, being non-invasive and safe. This investigation aimed to study protein turnover in healthy independent elderly through the method of glycine labeled with 15N. Seven healthy elderly persons were studied. All volunteers were assessed by clinic, nutritional and biochemical evaluation, with the exclusion of diseases and medications that could affect protein metabolism. A 200 mg oral dose of 15N-Glycine was administered and urine and blood samples were collected (basal sample before isotope intake, four hours after isotope intake and the last sample after 9 hours). Ammonium, urea and total nitrogen were quantified and analyzed by mass spectrometry, with the determination of isotope enrichment (15N). Volunteers were aged 65.4 ± 2.8 years (mean ± SE), four women and three men, with BMI 22.73 ± 2.4 Kg/m2. Total nitrogen output was 3.31 ± 0.7 gN/9hours and intake 7.76 ± 1.0 g/N; 15N nitrogen flux was 30.36 ± 6.3 gN/9hours, so the nitrogen balance was 4.46 ± 1.0 g/N. These findings were similar to those of others studies with old persons in the literature and lower than those for younger persons. This research established the values of protein metabolism in healthy old persons during the ingestion of typical food (rice and meat) and will allow the development of further intervention studies.
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Borella, Junior. "Metabolismo fermentativo em raízes e nódulos de soja sob condições de hipoxia e pós-hipoxia." Universidade Federal de Pelotas, 2011. http://repositorio.ufpel.edu.br/handle/ri/2052.
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Previous issue date: 2011-10-10Plant metabolism is sensitively to changes in oxygen availability by waterlogging. The immediate consequences of oxygen depletion are the activation of anaerobic metabolism and a reduction in energy production. This study aimed to evaluate the fermentative metabolism in soybean [Glycine max (L.) Merrill] roots and nodules under hypoxia and post-hypoxia conditions. Two experiments were conducted in greenhouse conditions with soybean genotypes Fundacep 53 RR and BRS Macota grown in 3L pots containing vermiculite as substrate, in association with Bradyrhizobium elkanii. The treatments were applied when the plants reached the R2 stage (reproductive). Experiment I - the root system of plants was subjected to hypoxia by fitting a second pot without holes and flooded with nutrient solution diluted to one-third of the original, in order to keep a blade of 20 mm on the surface of the substrate. Plants were maintained flooded for eight days, and so the pot without drilling holes was removed for recovery assessment. Non-waterlogging plants were maintained as control (normoxic condition). The activities of fermentation enzymes, alcohol dehydrogenase (ADH), pyruvate decarboxylase (PDC) and lactate dehydrogenase (LDH) were quantified in roots and nodules collected at 2, 4, 6 and 8 days after flooding or recovery . Experiment II - soybean plants were transferred to a hydroponic system and roots were subjected to hypoxia by flowing N2 gas during 24 and 72 h. For recovery, plants returned to vermiculite for 24 and 72 h. A control group of plants was maintained in vermiculite.The enzymatic activities of ADH, PDC and LDH, levels of anaerobic metabolites (ethanol, lactate and pyruvate) and carbohydrate content (sucrose, soluble sugars, starch and water soluble polysaccharides) were quantified in roots and nodules. The experiments were completely randomized with four replications and data were analyzed by ANOVA and compared by the Tukey test (P 0.05). Experiment I - activities of ADH, PDC and LDH increased significantly in roots and nodules of flooded plants and decreased during recovery. Fundacep 53 RR responded more effectively to treatments effects. Experiment II - the activity of ADH, PDC and LDH enzymes, as well as the content of ethanol and lactate, increased with hypoxia in roots and nodules, and returned to pre-hypoxic levels with the recovery in both genotypes. The pyruvate content increased in nodules and decreased in roots, sugars and sucrose levels increased in roots and decreased in nodules under hypoxia for both genotypes. Fundacep RR 53 was more responsive to the effects caused by hypoxia and post-hypoxia than BRS Macota. The effects were more pronounced in plants subjected to hypoxia by hydroponic system due to the rapid depletion of oxygen in the solution.
O metabolismo das plantas reage sensivelmente às mudanças de disponibilidade de oxigênio impostas pelo alagamento. Uma das consequências imediatas da depleção do oxigênio é a ativação do metabolismo anaeróbico e uma redução significativa na produção de energia. Este trabalho teve por objetivo avaliar o metabolismo fermentativo em raízes e nódulos de soja [Glycine max (L.) Merril] sob condições de hipoxia e pós-hipoxia. Para isso dois experimentos foram conduzidos em casa de vegetação com os genótipos Fundacep 53 RR e BRS Macota, cultivados em vasos de 3L contendo vermiculita como substrato, em associação com Bradyrhizobium elkanii. Os tratamentos foram aplicados quando as plantas atingiram o estádio R2 (reprodutivo). Experimento I - o sistema radicular da soja foi submetido à hipoxia pelo encaixe de um segundo vaso sem perfurações e inundado com solução nutritiva diluída a 1/3 da concentração original, de forma a se manter uma lâmina de 20 mm sobre a superfície do substrato. A inundação das plantas foi conduzida por 8 dias, após este período foi retirado os vasos sem perfurações para avaliação da recuperação. O material vegetal foi coletado no 2º, 4º, 6º e 8º dia após a inundação (DAI) ou recuperação (DAR), juntamente com seus respectivos controles (mantidos em normoxia) para a dosagem da atividade das enzimas fermentativas, álcool desidrogenase (ADH), piruvato descarboxilase (PDC) e lactato desidrogenase (LDH) em raízes e nódulos. Experimento II - as plantas foram transferidas para um sistema hidropônico e submetidas à hipoxia do sistema radicular pelo borbulhamento de N2 gasoso por 24 e 72 h ou mantidas em vermiculita (controle). Posteriormente retornaram para vermiculita sob condições de normoxia por 24 e 72 h. Foram avaliadas a atividade das enzimas ADH, PDC e LDH , metabólitos anaeróbicos (etanol, lactato e piruvato) e conteúdo de carboidratos (sacarose, açúcares solúveis totais, amido e polissacarídeos solúveis em água) em raízes e nódulos. O delineamento foi inteiramente casualizado com quatro repetições. Os dados foram submetidos à ANOVA e as médias comparadas pelo teste de Tukey a 5% de probabilidade. No experimento I a inundação promoveu aumento significativo da atividade da ADH, PDC e LDH, bem como a diminuição da atividade com a recuperação em raízes e nódulos. Fundacep 53 RR respondeu de forma mais efetiva aos tratamentos. No experimento II, a atividade das enzimas ADH, PDC e LDH, bem como os teores de etanol e lactato, aumentaram com a hipoxia em raízes e nódulos e retornaram aos níveis pré-hipóxicos com a recuperação em ambos os genótipos. O teor de piruvato aumentou nos nódulos e diminuiu nas raízes e os teores de açúcares e sacarose aumentaram nas raízes e diminuíram nos nódulos em hipoxia para ambos os genótipos. Fundacep 53 RR foi mais responsiva aos efeitos provocados pela hipoxia e pós-hipoxia que BRS Macota. Os efeitos foram mais pronunciados nas plantas submetidas à hipoxia pelo sistema hidropônico em função da rápida depleção do oxigênio na solução.
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Lima, Juliana Domingues. "Transporte de nitrogenio e metabolismo da aspargina em soja (Glycine Max L.) sob deficiencia na assimilação do nitrogenio." [s.n.], 2002. http://repositorio.unicamp.br/jspui/handle/REPOSIP/315243.
Full textAbstract:
Orientador: Ladaslav SodekTese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: Foram conduzidos experimentos com plantas de soja e outras leguminosas em condições de casa-de-vegetação. Plantas noduladas e não-noduladas apresentaram aumento significativo do nível de ácido aspártico e redução da asparagina transportada na seiva do xilema sob condições que limitam a assimilação do nitrogênio. Em soja, a resposta foi revertida quando as condições ótimas para assimilação do nitrogênio foram restauradas. Plantas não-noduladas, crescidas em vasos contendo vermiculita, usando nitrato como fonte de nitrogênio, apresentaram este fenômeno quando transferidas para hidroponia em solução deficiente em nitrogênio. Subseqüentemente, quando nitrato foi novamente fornecido o efeito foi revertido. Plantas noduladas crescidas em vermiculita recebendo solução deficiente em nitrogênio e totalmente dependente da fixação do N2 como fonte de nitrogênio apresentaram a mesma resposta quando transferi das para hidroponia com aeração. Neste caso, após a transferência para hidroponia os níveis de ureídeos na seiva do xilema decresceram significativamente sugerindo, com outras evidências, que a fixação do nitrogênio foi prejudicada nessas condições. Com o retomo destas plantas para vermiculita, ambos a razão ASPI ASN e o nível de ureídeos retomaram aos valores iniciais. Durante o crescimento e a nodulação de plantas de soja cultivadas em vermiculita em solução nutritiva deficiente em nitrogênio, o fenômeno foi observado durante curto período que corresponde à fase de "fome de N". Esta fase é caracterizada pelo transitório amarelecimento das folhas e ocorre quando as reservas são exauridas e a nodulação não está suficientemente desenvolvida para manter a planta com nitrogênio fixado. Seguindo os tratamentos descritos acima, a análise dos aminoácidos livres do floema, raízes e nódulos também revelou aumento da razão ASP/ASN mas, muito menos pronunciado do que o encontrado na seiva do xilema. A atividade da asparagina sintetase foi investigada no sistema radicular para tentar elucidar os fatores envolvidos no fenômeno. A atividade da enzima nos nódulos sofreu uma queda nos tratamentos que elevaram a razão ASPI ASN como a transferência das plantas noduladas para hidroponia e a queda foi recuperada com o retomo das plantas para vermiculita quando a razão ASP/ASN atingiu o nível normal. Entretanto, houve insucesso na dosagem da atividade da asparagina sintetase nas raízes. Evidentemente, em termos qualitativos, o transporte de nitrogênio na seiva do xilema parece ser um indicador bastante sensível do status de nitrogênio da planta
Abstract: Experiments with soybean plants and other legumes were conducted under greenhouse conditions. Both nodulated and non-nodulated plants were found to present substantially increased aspartic acid levels and lower asparagine in the xylem bleeding sap under conditions that limit nitrogen assimilation. In soybean, the response was reversed when optimum conditions for nitrogen assimilation were restored. Non-nodulated plants, grown in pots with vermiculite using nitrate as sole source of nitrogen, presented this phenomenon when transferred to N-deficient water-culture. Subsequent return to nitrate reversed the effect. Nodulated plants grown in vermiculite with N-deficient nutrient solution and totally dependent on N2-fixation as a source of nitrogen presented the same response when transferred to aerated water -culture. In this case xylem ureide levels also fell substantially suggesting, along with other evidence, that nitrogen fixation was impaired under these conditions. On return of plants to vermiculite, both the Asp/Asn ratios and ureide levels returned to initial values. During growth and nodulation of soybean plants cultivated in vermiculite with N-deficient nutrient solution, the phenomenon was observed over a short period corresponding to the "N hunger" phase. This phase is characterised by transient yellowing of the leaves and occurs when reserves are depleted and nodulation is not yet sufficiently developed to maintain the plant with fixed nitrogen. Following the above-described treatments, an analysis of free amino acids in the phloem, roots and nodules also revealed some increase in the Asp/ Asn ratios but these were much less pronounced than those found for the xylem. Asparagine synthetase activity was investigated in the root system in an attempt to elucidate the factors underlying the phenomenon. Activity in the nodules was found to fall during treatments which elevated the Asp/Asn ratio such as transfer of nodulated plants to water-culture and the fall in activity reversed on return of the plants to vermiculite when ratios returned to normal. However, measurement of asparagine synthetase activity in the roots was unsuccessful. Evidently, in qualitative terms, nitrogen transport in the xylem appears to be very sensitive to the nitrogen status of the plant
Doutorado
Doutor em Biologia Vegetal
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Borba, Isabel Cristina Gouvêa de. "Metabolismo Antioxidativo para a classificação de lotes de sementes quanto ao vigor." Universidade Federal de Pelotas, 2013. http://repositorio.ufpel.edu.br/handle/ri/1998.
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Previous issue date: 2013-05-24The increased demand for high quality seeds led seed technology companies to improve their techniques aiming to associate germination and vigor with the responses of the seeds in the field. However, such responses may suffer several influences during storage and maturation process, bringing damage to the membranes' systems, leading to reactive oxygen species (ROS), which can diminish seed vigor. Thus, the antioxidative system enzymes may become an enriching alternative to classify seeds lots according to their vigor. Most works analyzing those enzymes activity report their responses related to the abiotic stress influence in different crops. In such cases, the decrease in the enzymatic activity point to seeds and seedlings with less vigor and viability. However, in this work, where the seeds were not submitted to any kind of stress, those enzymes higher activity expressed a higher ROS production, and that points to the beginning of the deterioration process, featuring lesser seeds vigor. Considering this, the work aimed to distinguish seeds lots of maize, beans and soybean through the antioxidative enzymes activity (SOD superoxide dismutase, APX ascorbate peroxidase and CAT - catalase), and relate them to viability and vigor standard test to verify if those enzymes detect differences in vigor of seeds lots. For this, the seeds of the three crops were submitted to the physiological quality standard tests, such as germination, first count and germination speed index, length and dry mass of roots and shoots, electrical conductivity and antioxidative enzymes. Through the obtained results, it was possible to verify that in beans and soybean seeds lots, which presented high heterogeneity between them, the antioxidative system enzymes activity was not efficient to separate the lots. However, in the maize seeds lots that presented high homogeneity for the physiological quality standard tests, it was possible identifying differences in the enzymes activity in the seedlings derived from those seeds lots, allowing to classify them into different vigor levels.
O aumento da demanda por sementes de alta qualidade levou as empresas de tecnologia de sementes a aprimorarem suas técnicas visando associar os testes de geminação e vigor com as respostas das sementes no campo. Contudo, tais respostas podem ser influenciadas durante o processo de maturação e armazenamento, acarretando danos aos sistemas de membranas e levando a formação de espécies reativas de oxigênio (EROs), as quais podem levar a diminuição do vigor das sementes. Deste modo, a avaliação das enzimas do sistema antioxidativo pode se tornar uma alternativa enriquecedora para classificar lotes de sementes quanto ao vigor. A maioria dos trabalhos que analisam a atividade dessas enzimas relacionam suas respostas frente a influência de estresses abióticos em diferentes culturas, nesses casos o decréscimo da atividade enzimática remete sementes e plântulas de menor viabilidade e vigor, contudo, neste trabalho onde as sementes não foram submetidas a nenhum tipo de estresse a maior atividade dessas enzimas representaria a maior produção de EROs que, consequentemente, evidencia o início do processo deteriorativo, caracterizando o menor vigor das sementes. Diante disso, neste trabalho o objetivo neste trabalho foi diferenciar lotes de sementes de milho, feijão e soja por meio da atividade das enzimas antioxidantes (SOD superóxido dismutase, APX ascorbato peroxidase e CAT catalase) e verificar se estas enzimas detectam pequenas diferenças no vigor das sementes. Para isso, as sementes das três culturas foram submetidas aos testes padrão de qualidade fisiológica, como germinação, primeira contagem e índice de velocidade de germinação, comprimento e massa seca da parte aérea e raízes, condutividade elétrica e posteriormente, submetidas à análise da atividade de enzimas antioxidantes. Através dos resultados obtidos foi possível verificar que nos lotes das sementes de feijão e soja, que apresentaram alta heterogeneidade entre eles a atividade das enzimas do sistema antioxidativo não foi eficiente na separação dos lotes, contudo, nos lotes de sementes de milho que apresentaram elevada homogeneidade pelos testes padrão de qualidade fisiológica foi possível evidenciar diferença na atividade das enzimas das plântulas oriundas desses lotes de sementes, permitindo classificar os mesmos em diferentes níveis de vigor.
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Martin, Leonardo Curi [UNESP]. "Identificação de SNPs (Single Nucleiotide Polymorphisms) no gene colina monooxigenase relacionado ao metabolismo da glicina betaína em Eucalyptus." Universidade Estadual Paulista (UNESP), 2010. http://hdl.handle.net/11449/92452.
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Estresses abióticos, como a seca, podem reduzir significativamente os rendimentos no cultivo de espécies comercialmente importantes, tais como o eucalipto na produção de celulose e papel. Quando submetidas às condições de déficit hídrico, as plantas desenvolvem alguns mecanismos de defesa. As betaínas participam destes mecanismos, sendo seu soluto mais comum a glicina betaína. Esta é uma amina quaternária distribuída extensamente em diversas espécies de plantas superiores, sintetizada em elevadas taxas, tendo como função, manter a turgescência celular. A identificação e estudo de genes relacionados à tolerância à seca são importantes para os programas de melhoramento florestal. Este estudo teve por objetivo identificar SNPs (Single Nucleotide Polymorphisms) no gene colina monooxigenase relacionado ao metabolismo da glicina betaína em Eucalyptus. A sequência do gene em estudo foi encontrada em genomas de plantas modelos através do banco de dados GenBank, sendo as mesmas, utilizadas para a procura de similaridades no banco de dados de ESTs de eucalipto FORESTs – FAPESP. Após a constatação de homologia no banco de dados de eucalipto, foram confeccionados oito pares de “primers” para flanquear essas regiões, sendo estes, avaliados, amplificando fragmentos únicos. Depois de realizado o seqüenciamento do gene colina monooxigenase, foi utilizado a ferramenta BLAST no GenBank, confirmando com sucesso a identidade da sequência. Em seguida, as sequências foram alinhadas e os SNPs encontrados. No total, foram identificados 49 SNPs, sendo 12 em regiões codificantes e 37 em regiões UTRs e íntrons. Somente os SNPs localizados nas regiões codificantes foram utilizados neste trabalho, sendo 83,3% deles possuindo mutações sinônimas e 16,7% não-sinônimas. Em seguida, foi utilizada uma população mais abrangente composta de E. grandis...
Abiotic stress, such as drought, can reduce significantly the yield in the important commercially species crop, such as the eucalyptus in the cellulose and paper. When submitted to the water stress conditions, the plants develop some defense mechanisms. The betaines are part of these mechanisms , being their solute more common to the glycine betaine. This is a quaternary amine extensively distributed in several species of higher plants synthesized in high taxes with the function of maintaining the cellular turgescence. Identifying and studying the genes related to the drought tolerance are important for the forest improvement programs. This work aimed at identifying SNPs (Single Nucleotide Polymorphisms) in choline monooxygenase related to the glycine betaine in Eucalyptus. The gene sequence was found in model plants genomes through the databank GenBank, being used for searching similarities in the databank ESTs of eucalyptus FORESTs – FAPESP. After noticing homology in the eucalyptus databank, eight pairs of primers were made to amplify these regions, being evaluated, amplifying unique fragments. After the choline monooxygenase sequencing was performed, it was used the BLAST in the GenBank, confirming successfully the sequence identity. Then, the sequences were aligned and the SNPs were found. In the total, 49 SNPs were identified, being 12 in coding regions and 37 in UTRs and intron regions. Only the SNPs located in coding regions were used in this work, being 83.3% of the SNPs with synonymous mutation and 16.7% non-synonymous. Following, it was used a wider population made up of de E. grandis, E. Urophylla and the hybrid “Urograndis” to perform the SNPs genotyping, establishing the formation of 18 haplotypes and 16 possible haplotypical combinations which revealed that some genotypes were exclusive for the species E. grandis and E. urophylla. With the objective of decreasing costs... (Complete abstract click electronic access below)
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Martin, Leonardo Curi. "Identificação de SNPs (Single Nucleiotide Polymorphisms) no gene colina monooxigenase relacionado ao metabolismo da glicina betaína em Eucalyptus /." Botucatu : [s.n.], 2010. http://hdl.handle.net/11449/92452.
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Orientador: Celso Luís MarinoBanca: Rinaldo César de Paula
Banca: Ivan de Godoy Maia
Resumo: Estresses abióticos, como a seca, podem reduzir significativamente os rendimentos no cultivo de espécies comercialmente importantes, tais como o eucalipto na produção de celulose e papel. Quando submetidas às condições de déficit hídrico, as plantas desenvolvem alguns mecanismos de defesa. As betaínas participam destes mecanismos, sendo seu soluto mais comum a glicina betaína. Esta é uma amina quaternária distribuída extensamente em diversas espécies de plantas superiores, sintetizada em elevadas taxas, tendo como função, manter a turgescência celular. A identificação e estudo de genes relacionados à tolerância à seca são importantes para os programas de melhoramento florestal. Este estudo teve por objetivo identificar SNPs (Single Nucleotide Polymorphisms) no gene colina monooxigenase relacionado ao metabolismo da glicina betaína em Eucalyptus. A sequência do gene em estudo foi encontrada em genomas de plantas modelos através do banco de dados GenBank, sendo as mesmas, utilizadas para a procura de similaridades no banco de dados de ESTs de eucalipto FORESTs - FAPESP. Após a constatação de homologia no banco de dados de eucalipto, foram confeccionados oito pares de "primers" para flanquear essas regiões, sendo estes, avaliados, amplificando fragmentos únicos. Depois de realizado o seqüenciamento do gene colina monooxigenase, foi utilizado a ferramenta BLAST no GenBank, confirmando com sucesso a identidade da sequência. Em seguida, as sequências foram alinhadas e os SNPs encontrados. No total, foram identificados 49 SNPs, sendo 12 em regiões codificantes e 37 em regiões UTRs e íntrons. Somente os SNPs localizados nas regiões codificantes foram utilizados neste trabalho, sendo 83,3% deles possuindo mutações sinônimas e 16,7% não-sinônimas. Em seguida, foi utilizada uma população mais abrangente composta de E. grandis... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: Abiotic stress, such as drought, can reduce significantly the yield in the important commercially species crop, such as the eucalyptus in the cellulose and paper. When submitted to the water stress conditions, the plants develop some defense mechanisms. The betaines are part of these mechanisms , being their solute more common to the glycine betaine. This is a quaternary amine extensively distributed in several species of higher plants synthesized in high taxes with the function of maintaining the cellular turgescence. Identifying and studying the genes related to the drought tolerance are important for the forest improvement programs. This work aimed at identifying SNPs (Single Nucleotide Polymorphisms) in choline monooxygenase related to the glycine betaine in Eucalyptus. The gene sequence was found in model plants genomes through the databank GenBank, being used for searching similarities in the databank ESTs of eucalyptus FORESTs - FAPESP. After noticing homology in the eucalyptus databank, eight pairs of primers were made to amplify these regions, being evaluated, amplifying unique fragments. After the choline monooxygenase sequencing was performed, it was used the BLAST in the GenBank, confirming successfully the sequence identity. Then, the sequences were aligned and the SNPs were found. In the total, 49 SNPs were identified, being 12 in coding regions and 37 in UTRs and intron regions. Only the SNPs located in coding regions were used in this work, being 83.3% of the SNPs with synonymous mutation and 16.7% non-synonymous. Following, it was used a wider population made up of de E. grandis, E. Urophylla and the hybrid "Urograndis" to perform the SNPs genotyping, establishing the formation of 18 haplotypes and 16 possible haplotypical combinations which revealed that some genotypes were exclusive for the species E. grandis and E. urophylla. With the objective of decreasing costs... (Complete abstract click electronic access below)
Mestre
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Sousa, Carlos Antonio Ferreira de. "Metabolismo de nitrogenio em plantas de soja [Glycine max (L.) Merr. cv IAC-17] submetidas a deficiencia de O2 no sistema radicular." [s.n.], 2001. http://repositorio.unicamp.br/jspui/handle/REPOSIP/315234.
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Orientador: Ladaslav SodekTese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: A soja [Glycine max (L.) Merr.] é um dos modelos mais usados para se estudar a deficiência de 'O IND.2¿ no sistema radicular de plantas. Apesar disso, a literatura é carente de informações a respeito do encharcamento radicular na soja e seus efeitos sobre os teores de metabólitos, especialmente os nitrogenados, nas raízes e suas formas de transporte pelo xilema e floema. Portanto, este trabalho teve os seguintes objetivos: 1. determinar os efeitos da hipoxia no sistema radicular da soja sobre os teores de metabólitos nas raízes, nos exsudatos do xilema e floema e sobre a atividade das enzimas alanina aminotransferase e aspartato aminotransferase nas raízes; 2. determinar o tempo necessário para a recuperação dos teores dos metabólitos nas raízes e no exsudato do xilema e da atividade da enzima alanina aminotransferase em raízes de soja, após o retorno à normoxia; 3. Purificar e caracterizar a enzima alanina aminotransferase em raízes de soja sob hipoxia. Plantas de soja cv IAC-17 não noduladas foram cultivadas em solução nutritiva sob normoxia (borbulhamento contínuo de ar comprimido) e, ao atingir o estádio V5, foram submetidas aos seguintes experimentos: 1. Entradaem hipoxia: suspensão do borbulhamento de ar comprimido, combinado com a aplicação de uma camada de óleo mineral sobre a solução por até 120 h; 2. Hipoxia seguida pelo retorno à normoxia: suspensão do borbulhamento de ar comprimido, combinado com o borbulhamento de 'N IND.2¿ gasoso na solução por 120 h, com posterior retorno à normoxia por até 96 h. Em ambos os experimentos, sob normoxia, os teores de 'O IND.2¿ na solução nutritiva situaram-se em torno de 87% de saturação (6,5 mg/ml). Ao se iniciar a hipoxia no experimento 1, houve uma diminuição gradativa nos teores de 'O IND.2¿,que se estabilizaram em torno de 6% de saturação (0,4 mg/ml), depois de 8 h, enquanto no experimento 2, houve uma diminuição abrupta nos teores de 'O IND.2¿ na solução nutritiva, após 6 h de hipoxia, para valores em torno de 5% (0,3 mg/ml) e, após 24 h, para 4% (0,2 mg/ml). Nas raízes de soja, observaram-se aumentos nos teores de amido e WSP nas primeiras 24 h de hipoxia, nos teores de aminoácidos até 48 h, e nos teores de açúcares solúveis totais, sacarose e proteína, a partir de 48 h até 96 h de hipoxia. Houve diminuição nos teores de 'N0 IND.3¿ ao longo de 96 h de hipoxia. No exsudato do xilema, houve aumento nos teores de aminoácidos, também, até 48 h de hipoxia, mas uma diminuição abrupta nos teores de 'N0 IND.3¿ nas primeiras 24 h. No exsudato do floema, houve uma redução pela metade nos teores de aminoácidos, açucares solúveis totais e sacarose, após 96 h de hipoxia, comparando-se ao controle normóxico. No retorno à normoxia, o restabelecimento dos teores de metabólitos encontrados nas raízes e no exsudato do xilema, no período anterior à hipoxia, ocorreu dentro de 48 h, com exceção de 'NO IND.3¿ e proteína, cujos teores foram restabelecidos aos níveis do controle, após 72 h. Houve aumentos nos teores de Ala e lactato em raízes de soja nas primeiras 48 h de hipoxia, e de piruvato e etanol, até 120 h de hipoxia. Sob hipoxia radicular, o destino principal do piruvato foi.sua conversão a Ala, que se acumulou na proporção de 5:1 e 6:1, em relação a lactato e etanol, respectivamente. No retorno à normoxia, as raízes de soja recuperaram os teores pré-hipóxicos dos metabólitos da anaerobiose em 48 h
Abstract: The soybean (Glycine max (L.) Merr.) is among the most frequently used models to study 'O IND.2¿ deficiency in the root system of plants. However, few studies have focussed on waterlogging of the root system and its effects on the metabolic content of the roots, especially nitrogenous compounds, and their means of transportation through the xylem and phloem. The objectives of this study were: 1) assess the effects of hypoxia of the soybean root system on the content of important metabolites of roots, xylem and phloem sap, together with the activity of the enzymes alanine aminotransferase and aspartate aminotransferase in the roots; 2) determine the time required for the re-establishment of the levels of such metabolites in the roots and in the xylem exsudate as well as alanine aminotransferase activity in the reots, after the return to normoxia; 3) partially purify and characterise the alanine aminotransferase enzyme in soybean roots under hypoxia. Non-nodulated cv IAC -17 soybean plants were cultivated in nutritive solution under normoxia (continuous aeration by compressed air) and, after reaching the V5 stage, they were used in the following basic experimental systems: a) Hypoxia: interruption of aeration together with the application of a layer of mineral oil over the solution, for up to 120 h; b) Hypoxia followed by the return to normoxia: solution aerated with 'N IND.2¿ gas for 120 h followed by the restoration of aeration by compressed air for 96 h. Under normoxia, the 'O IND.2¿ content in the nutritive solution was around 87% saturation (6,5 mg/ml) for both experimental systems. Under conditions of the first experimental system, the 'O IND.2¿ content fell gradually and after 8 h of hypoxia stabilised at 6% saturation (0,4 mg/ml). This decrease was more abrupt in the second experimental system, where the 'O IND.2¿ content dropped to around 5% (0,3 mg/ml) after 6 h and 4 % (0,2 mg/ml) after 24 h of hypoxia. During hypoxia, the starch and water soluble polissacharids contents of the roots showed an increase after 24 h, amino acids after 48 h and total soluble sugar, sucrose and protein from 48 to 96 hours. The 'N0 IND.3¿ content decreased after 96 h of hypoxia. In the xylem exsudate, amino acid levels increased during 48 h of hypoxia, while 'N0 IND.3¿ decreased abruptly within the first 24 h. Phloem exsudates presented a 50% decrease of amino acids, total soluble sugar and sucrose after 96 h of hypoxia, compared to normoxia values. After normoxia was re-established, the metabolite content of the roots and xylem exsudates were normal after 48 h, except for 'N0 IND.3¿- and protein, which reached control levels after 72 hours. The content of Ala and lactate in the soybean roots increased within 48 h of hypoxia while pyr and ethanol increased up to 120 h. It appears that under hypoxia, the principal metabolic fate of pyr was its conversion to Ala, which accumulated in the proportion of 5:1 and 6:1 in relation to lactate and ethanol, respectively. After returning to normoxia, the roots recovered the levels of these metabolites to those observed before the hypoxia within 48 hours. Under normoxia, Asn was the predominant amino acid in the roots and xylem exsudate. However, during root hypoxia, Asn decreased sharply while the content of Ala and Gaba rose dramatically.When normoxia was re-established, these amino acids reached their original levels within 48 h
Doutorado
Doutor em Biologia Vegetal
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Pocard, Jean-Alain. "La glycine bétaïne : effet osmoprotecteur, transport et métabolisme chez rhizobium meliloti en culture in vitro et en symbiose avec medicago sativa L." Rennes 1, 1987. http://www.theses.fr/1987REN10060.
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Antunes, Flavia. "Papel da redutase do nitrato e da asparagina sintetase em plantas de soja (Glycine max L.) sob condições de estresse de nitrogenio." [s.n.], 2007. http://repositorio.unicamp.br/jspui/handle/REPOSIP/315227.
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Orientador: Ladaslav SodekTese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: O sistema radicular da soja (Glycine max L.) é um importante sítio da assimilação do nitrogênio (N), seja pela assimilação do nitrato nas raízes, seja pela fixação simbiótica de N atmosférico nos nódulos. Os principais produtos da assimilação do N inorgânico, os aminoácidos asparagina e glutamina, os ureídeos, a alantoína e o ácido alantóico, são usados no transporte de N para a parte aérea. Assim, esses produtos representam uma fonte de N reduzido tanto para os sítios de consumo, quanto para a formação de outros aminoácidos, proteínas, ácidos nucléicos e dos demais compostos nitrogenados sintetizados na célula. O transporte do N assimilado no sistema radicular para a parte aérea é realizado exclusivamente via xilema. Portanto, o objetivo geral deste trabalho foi, relacionar mudanças na composição de aminoácidos transportados no xilema de plantas de soja, provocadas pela deficiência de N, com o comportamento de enzimas de assimilação do N, encontradas no sistema radicular. Para causar a deficiência do N, as plantas de soja foram transferidas para um meio hidropônico sem nitrato ou qualquer outra fonte de N inorgânico. Nas plantas não-noduladas a deficiência se deu pela ausência do nitrato no meio de cultivo, e nas plantas noduladas, a manutenção em hidroponia inibiu a fixação do N2. Foi observado um aumento na razão aspartato/asparagina (ASP/ ASN) na seiva do xilema das plantas não-noduladas e noduladas, quando foram submetidas à deficiência do N. Contudo, nas plantas noduladas esse aumento foi acentuado apenas no primeiro dia do tratamento. A recuperação do estresse só ocorreu em plantas não-noduladas, nas quais as concentrações de ASP e ASN retomaram seus valores. As análises da expressão dos genes que codificam a asparagina sintetase (AS) no sistema radicular das plantas de soja, durante o , leva à uma diminuição dos níveis de glutamina, produto imediato da assimilação do íon amônio através da enzima glutamina sintetase (GS). Com a redução dos níveis de glutamina a atividade da AS também é reduzida, resultando na menor utilização de ASP. Em plantas não-noduladas, após transferência para o meio hidropônico sem nitrato, a expressão dos três genes para AS diminuiu bruscamente, e após o retorno das plantas ao meio com nitrato, os genes da AS foram expressos novamente. Nas plantas noduladas a expressão gênica da AS também reduziu durante o tratamento de deficiência de N, porém, apenas o gene SAS1 parece ter sido afetado. O gene SAS1 não recuperou seus níveis de expressão durante a tentativa de recuperação do estresse. Este fato é mais uma evidência da relação da AS com as alterações no perfil de aminoácidos transportados na seiva do xilema, pois as concentrações de ASN e ASP na seiva do xilema também não foram retomadas. tratamento e recuperação do estresse de N, indicam que essa enzima está relacionada com as alterações na razão ASP/ASN. A AS é a enzima que catalisa a biossíntese da ASN, transferindo o grupo amino da glutamina para o aspartato, produzindo asparagina e glutamato. A queda no suprimento de N, seja pelo impedimento da assimilação do nitrato ou da fixação do N2. Devido à importância da enzima redutase do nitrato (RN) no processo de assimilação do nitrato pelo sistema radicular, foi avaliado o seu comportamento durante o tratamento de deficiência do nitrato e sua recuperação. A atividade da RN aumentou, consideravelmente, quando as plantas foram expostas ao nitrato. Esse aumento pôde ser observado em raízes de plantas de soja não-noduladas e também no sistema radicular (raízes e nódulos) das plantas noduladas. Quando as plantas não-noduladas foram transferidas para uma solução sem nitrato, a atividade da RN caiu, apresentando valores muito baixos dentro de 24 horas. As atividades da RN ) parece bastante clara. De qualquer forma, a forte dependência da RN da presença do nitrato é um dado inédito para raízes de plantas de soja. O nitrato também afetou a expressão gênica da AS nas raízes de plantas noduladas, cultivadas sem nenhuma fonte de N mineral. A expressão dos genes da AS aumentou quando essas plantas receberam solução contendo nitrato. nas raízes e nódulos de plantas de soja, cultivadas na ausência do nitrato, foram muito baixas, sendo que em nódulos a atividade sempre foi ligeiramente maior que nas raízes. A baixa atividade encontrada nas raízes e nódulos de soja pode representar a forma constitutiva da enzima, pois as plantas foram cultivadas sem a adição do nitrato durante o ciclo todo. Entretanto, não pode ser descartada a possibilidade da presença de traços de nitrato na água usada para regar as plantas e isto ter sido suficiente para induzir a baixa atividade encontrada. Se por um lado podem existir dúvidas quanto ao fato da enzima ser constitutiva, a indução da enzima pelo substrato (NO3- Os dados sobre o comportamento das enzimas avaliadas aqui indicam que as alterações em aminoácidos transportados no xilema, em plantas de soja submetidas à deficiência de N, estão relacionadas com os processos assimilatórios do sistema radicular, e que a AS parece ser a principal responsável pelas alterações na razão ASP/ASN
Abstract: The root system of soybean (Glycine max L.) is an important site for the assimilation of nitrogen, whether by nitrate assimilation in the roots, or by symbiotic nitrogen fixation in the nodules. The main products of inorganic nitrogen assimilation, the amino acids asparagine and glutamine and the ureides allantoin and allantoic acid, are used in the transport of assimilated nitrogen to the shoot. Thereby, these products represent a source of reduced nitrogen for the sink tissues, for the formation of other amino acids, proteins, nucleic acids and all the other nitrogenous compounds synthesized in the cell. Therefore, the objective of this study was to relate changes in the transport of nitrogen in the xylem of soybean caused by nitrogen deficiency with the behaviour of certain enzymes of nitrogen assimilation in the root system. Nitrogen deficiency was induced by the transfer of soybean plants to a hydroponic system without nitrate or any other source of inorganic nitrogen. In the case of non-nodulated plants deficiency was imposed by the interruption of nitrate assimilation by the roots, and for the nodulated plants nitrogen fixation was inhibited by immersion of the nodules in the hydroponic system. Under nitrogen deficiency, an increase in the aspartate/asparagine (ASP/ASN) ratio of the xylem sap was observed in both nodulated and non-nodulated plants. Nevertheless, this increase was substantial only on the first day of treatment. The recovery from the stress was only observed for the non-nodulated plants, where the levels of ASP and ASN returned to their initial values. Analyses of asparagine synthetase expression in the root system of soybean during treatment and recovery from nitrogen stress indicates that this enzyme can underlay the changes in ASP/ASN ratios. AS is an enzyme that catalyses the biosynthesis of Asn, by transferring the amide nitrogen from glutamine to aspartate, producing asparagine and glutamate. The fall in N supply, whether by interruption of nitrate assimilation or nitrogen fixation, leads to a decline in glutamine, the immediate product of ammonium ion assimilation via glutamine synthetase. With the reduction in glutamine levels the activity of AS is also reduced resulting in diminished utilization of ASP. In non-nodulated plants, after the transfer to the hydroponic system without nitrate, the expression of the three genes declines sharply, and after the return of the plants to a supply of nitrate the AS genes are expression again. In nodulated plants the expression of AS was also reduced during treatment, however, in this case only the gene SAS1 was affected. The SAS1 gene did not recover its initial levels of expression after removing the stress which is further evidence for the correlation between AS activity and the changes in ASP/ASN ratios in the xylem sap, since these ratios were also not recovered. In view of the importance of nitrate reductase (NR) in the process of nitrate assimilation by the root system, its behaviour was evaluated during nitrate deficiency and recovery. The activity of NR increased considerably when plants were supplied with nitrate, in the case of roots of non-nodulated as well as the root system (roots and nodules) of nodulated plants. When non-nodulated plants were transferred to a nutrient solution free of nitrate, NR activity fell sharply, almost disappearing within 24 hours. RN activities in roots and nodules of soybean grown in the absence of nitrate were very low, with activity in the nodules being somewhat higher than in the roots. The low activity found in the roots and nodules could be due to a constitutive enzyme since plants were grown throughout with nitrate-free medium. However, the possible presence of trace amounts of nitrate in the tap water used to irrigate the plants cannot be discarded and may have been sufficient to induce the low levels of enzyme found. If on the one hand there are doubts as to the presence of a constitutive enzyme, the presence of the induced form is very clear. In any case, the strong dependence of NR on the presence of nitrate is an unknown fact for soybean roots. Nitrate also affected the expression of AS in roots of nodulated plants grown without any mineral source of nitrogen, since its expression increased tremendously when such plants were supplied with nitrate. The data concerning the behaviour of the enzymes studied here indicate that alterations in xylem amino acids of soybean plants subjected to nitrogen deficiency are related to the assimilatory processes of the root system, and that AS appears to be mainly responsible for the changes in ASP/ASN ratios
Doutorado
Biologia Vegetal
Doutor em Biologia Vegetal
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Sugiyama, Yuta. "Biochemical studies and applications of sugar and polyamine metabolisms in gut microbes." Kyoto University, 2020. http://hdl.handle.net/2433/252995.
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Feitosa, Antonio Glaydson de Sousa. "Modulation of photosynthesis and nitrogen assimilation in conditions of high atmospheric CO2 on soybean plants." Universidade Federal do CearÃ, 2014. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=13167.
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CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel SuperiorHigh atmospheric CO2 concentrations limit the assimilation of CO2 and nitrogen in many species of C3 plants Thus the aim of this study was to identify how the high CO2 regulates photosynthesis and assimilation of nitrogen in soybean Therefore an experiment was conducted in growth chamber using soybean plants, 25 days old CO2 levels were applied environment ( 40 Pa) high (100 Pa) and transition to ambient conditions ( 100-40 Pa ) High CO2 was able to strongly affect trade gas with stomatal closure leading to sharp reductions in leaf transpiration (88%) and CO2 assimilation ( 48%) Recovering 69% of the initial value of net photosynthesis after the transition period at environmental conditions , however the internal CO2 concentration ( Ci) was increased by 17 times returning to the level of control after the transition the initial activity and the total Rubisco decreased 44 % and 34 % respectively Taking a 9% reduction in enzyme activation state but after the transition to the conditions of CO2 environment the initial activity and total recovered 74 % and 84 % respectively have the nitrate reductase showed a strong inhibition over that period compared to the ambient condition reaching a maximum reduction of 64 % after four days of exposure to elevated CO2 contrary this enzyme had recovered its initial capacity at 90 % after the transition period in conclusion we can say that photosynthesis and nitrate reductase activity are practically inhibited by high atmospheric CO2 mainly controlled by stomatal and metabolic limitations
Altas concentraÃÃes de CO2 atmosfÃrico limitam a assimilaÃÃo de CO2 e de nitrogÃnio em muitas espÃcies de plantas C3 Dessa forma o objetivo deste trabalho foi identificar como o elevado CO2 regula a fotossÃntese e a assimilaÃÃo do nitrogÃnio em plantas de soja Para tanto foi realizado um experimento em cÃmara de crescimento utilizando plantas de soja com 25 dias de idade Os nÃveis de CO2 aplicados foram ambiente (40 Pa) elevado (100 Pa) e transiÃÃo para as condiÃÃes ambiente (100 â 40 Pa) O elevado CO2 foi capaz de afetar fortemente as trocas gasosas com fechamento estomÃtico levando a fortes reduÃÃes na transpiraÃÃo foliar (88%) e assimilaÃÃo de CO2 (48%) Recuperando 69% do valor inicial da fotossÃntese lÃquida apÃs o perÃodo de transiÃÃo para a condiÃÃo ambiente Entretanto a concentraÃÃo interna de CO2 (Ci) foi elevada em 17 vezes retornando para o nÃvel de controle apÃs a transiÃÃo A atividade inicial e a total da Rubisco decresceram 44% e 34% respectivamente Levando a uma reduÃÃo de 9% no estado de ativaÃÃo da enzima porÃm apÃs a transiÃÃo para as condiÃÃes de CO2 ambiente a atividade inicial e a total recuperaram 74% e 84% respectivamente JÃ a redutase do nitrato apresentou uma forte inibiÃÃo ao longo desse perÃodo quando comparada com a condiÃÃo ambiente chegando a uma reduÃÃo mÃxima de 64% apÃs quatro dias de exposiÃÃo ao elevado CO2 Contrariamente esta enzima teve sua capacidade inicial recuperada em 90% apÃs o perÃodo de transiÃÃo Em conclusÃo podemos afirmar que a fotossÃntese e a atividade da redutase do nitrato sÃo praticamente inibidas por elevado CO2 atmosfÃrico controladas principalmente por limitaÃÃes estomÃticas e metabÃlicas
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FOUGERE, FRANCOISE. "Adaptation aux contraintes salines chez les bacteroides isoles de nodosites de medicago sativa l. : transport et metabolisme de la glycine betaine, accumulation d'autres solutes compatibles." Nice, 1991. http://www.theses.fr/1991NICE4470.
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Les bacteroides isoles de nodosites de medicago sativa (luzerne) et places en milieu de faible osmolarite transportent la glycine betaine tres lentement, par diffusion passive. Au contraire, en presence de nacl 0,2 m, la glycine betaine est transportee par un mecanisme actif, a forte affinite, probablement dependant de la force proton motrice. La vitesse maximale reste cependant plus faible (15 fois) que celle observee chez r. Meliloti en culture in vitro ou une proteine periplasmique affine pour la glycine betaine est caracterisee. Sa disparition chez le bacteroide pourrait expliquer la forte diminution de l'activite de transport. Les conditions requises pour la mesure in vitro de l'activite fixatrice d'azote des bacteroides (temps d'incubation court et pression d'oxygene reduite) sont incompatibles avec l'accumulation d'une quantite de glycine betaine suffisante pour assurer une restauration de l'ara en conditions de salinite elevee. Les bacteroides catabolisent activement la glycine betaine mais le catabolisme est bloque lors d'une addition de nacl. La penetration de choline s'effectue selon un transport actif a double affinite et, aussi, par diffusion passive. En milieu de faible osmolarite, la choline est rapidement metabolisee, essentiellement en derives phosphoryles. Par contre, en presence de nacl, une active synthese de glycine betaine est observee. Les activites des enzymes de biosynthese, la choline oxydase et la betainal deshydrogenase, ne sont que peu modifiees lors d'un apport de nacl. Les effets de contraintes salines sur la teneur en composes solubles des nodosites et des racines ont aussi ete analyses. Chez les bacteroides, l'equilibre osmotique est assure principalement par l'asparagine, le saccharose et le pinitol. En outre, les etudes microscopiques mettent en evidence un epaississement du cortex nodulaire, une accumulation d'amidon et une augmentation du volume de
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Feitosa, Antonio Glaydson de Sousa. "Modulação da fotossíntese e assimilação do nitrogênio por condições de elevado CO2 atmosférico em plantas de soja." reponame:Repositório Institucional da UFC, 2014. http://www.repositorio.ufc.br/handle/riufc/16892.
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FEITOSA, Antonio Glaydson de Sousa. Modulação da fotossíntese e assimilação do nitrogênio por condições de elevado CO2 atmosférico em plantas de soja. 2014. 71 f. : Dissertação (mestrado) - Universidade Federal do Ceará, Centro de Ciências Agrárias, Departamento de Ciências do Solo, Programa de Pós-Graduação em Agronomia - Solos e Nutrição de Plantas, Fortaleza-CE, 2014Submitted by Nádja Goes (nmoraissoares@gmail.com) on 2016-05-19T12:25:16Z No. of bitstreams: 1 2014_dis_agsfeitosa.pdf: 836229 bytes, checksum: f77f1e9d446fce346b7ef1c4f7d05eb7 (MD5)
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High atmospheric CO2 concentrations limit the assimilation of CO2 and nitrogen in many species of C3 plants Thus the aim of this study was to identify how the high CO2 regulates photosynthesis and assimilation of nitrogen in soybean Therefore an experiment was conducted in growth chamber using soybean plants, 25 days old CO2 levels were applied environment ( 40 Pa) high (100 Pa) and transition to ambient conditions ( 100-40 Pa ) High CO2 was able to strongly affect trade gas with stomatal closure leading to sharp reductions in leaf transpiration (88%) and CO2 assimilation ( 48%) Recovering 69% of the initial value of net photosynthesis after the transition period at environmental conditions , however the internal CO2 concentration ( Ci) was increased by 17 times returning to the level of control after the transition the initial activity and the total Rubisco decreased 44 % and 34 % respectively Taking a 9% reduction in enzyme activation state but after the transition to the conditions of CO2 environment the initial activity and total recovered 74 % and 84 % respectively have the nitrate reductase showed a strong inhibition over that period compared to the ambient condition reaching a maximum reduction of 64 % after four days of exposure to elevated CO2 contrary this enzyme had recovered its initial capacity at 90 % after the transition period in conclusion we can say that photosynthesis and nitrate reductase activity are practically inhibited by high atmospheric CO2 mainly controlled by stomatal and metabolic limitations
Altas concentrações de CO2 atmosférico limitam a assimilação de CO2 e de nitrogênio em muitas espécies de plantas C3 Dessa forma o objetivo deste trabalho foi identificar como o elevado CO2 regula a fotossíntese e a assimilação do nitrogênio em plantas de soja Para tanto foi realizado um experimento em câmara de crescimento utilizando plantas de soja com 25 dias de idade Os níveis de CO2 aplicados foram ambiente (40 Pa) elevado (100 Pa) e transição para as condições ambiente (100 – 40 Pa) O elevado CO2 foi capaz de afetar fortemente as trocas gasosas com fechamento estomático levando a fortes reduções na transpiração foliar (88%) e assimilação de CO2 (48%) Recuperando 69% do valor inicial da fotossíntese líquida após o período de transição para a condição ambiente Entretanto a concentração interna de CO2 (Ci) foi elevada em 17 vezes retornando para o nível de controle após a transição A atividade inicial e a total da Rubisco decresceram 44% e 34% respectivamente Levando a uma redução de 9% no estado de ativação da enzima porém após a transição para as condições de CO2 ambiente a atividade inicial e a total recuperaram 74% e 84% respectivamente Já a redutase do nitrato apresentou uma forte inibição ao longo desse período quando comparada com a condição ambiente chegando a uma redução máxima de 64% após quatro dias de exposição ao elevado CO2 Contrariamente esta enzima teve sua capacidade inicial recuperada em 90% após o período de transição Em conclusão podemos afirmar que a fotossíntese e a atividade da redutase do nitrato são praticamente inibidas por elevado CO2 atmosférico controladas principalmente por limitações estomáticas e metabólicas
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Fourmois, Laura. "Marquage métabolique de glycanes : diagnostic et approches thérapeutiques." Thesis, Université Paris-Saclay (ComUE), 2016. http://www.theses.fr/2016SACLS262.
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Cette thèse porte sur la méthode de marquage métabolique de glycanes. Elle consiste à utiliser des monosaccharides modifiés pouvant être métaboliquement introduits sur la membrane externe des cellules. Deux cibles ont été choisies, les bactéries Legionella pneumophila, et des cellules eucaryotes (lignée cellulaire du cancer de la prostate, PC3).Différents analogues saccharidiques ont été synthétisés pour les deux cibles portant différents rapporteurs chimiques, notamment des fonctions azoture, alcyne terminal, alcène terminal, cyclopropène et cétone. Une voie de synthèse commune a été développée mettant en jeu des dérivés d’esters de N-hydrosuccinimide et des dérivés saccharidiques amino, tels que la D-mannosamine, la D-galactosamine et le L-fucose pour les cellules eucaryotes et des analogues d’un précurseur de l’acide légionaminique pour les bactéries Legionella pneumophila.Des essais de marquages métaboliques de glycanes ont été effectués sur Legionella pneumophila. Pour cela, différents dérivés saccharidiques ont été incorporés par les bactéries, puis la révélation des rapporteurs chimiques avec des groupements complémentaires a été évaluée (azoture-cyclooctyne, alcène-tétrazine, cétone-hydrazide/alkoxyamine). La détection a été réalisée soit directement (fluorophore sur le partenaire), soit indirectement (reconnaissance d’un groupement biotine par une streptavidine portant un fluorophore). Les bactéries ont été ensuite observées par microscopie photonique et les observations ont mis en évidence un marquage membranaire.Pour les cellules eucaryotes (PC3), des essais de marquage métabolique ont été effectués afin de vérifier l’incorporation des monosaccharides modifiés via une détection par fluorescence. Une série d’outils portant les fonctions complémentaires aux rapporteurs chimiques, notamment un dérivé de cyclooctyne (TMDIBO) et un dérivé de tétrazine ont été synthétisés. Ils ont été couplés à des dérivés de biotine afin d’obtenir des outils pour la microscopie photonique et à des ARMs (Antibody Recruiting Molecules), tel que le 2,4-dinitrophényle et le rhamnose, en vue d’une potentielle approche thérapeutique. Celle-ci consiste à combiner l’incorporation de monosaccharides modifiés et leur réaction avec un partenaire portant des ARMs, afin de recouvrir la surface des cellules par ces motifs. Les ARMs en présence de sérum humain vont ensuite activer le complément conduisant à la lyse des cellules marquées. Des tests de marquage métabolique ont été réalisés avec les outils couplés aux ARMs et une détection par fluorescence a permis de vérifier la présence des ARMs à la surface des cellules. Des premiers essais ont été effectués avec du sérum humain et des optimisations sont à réaliser.Le marquage métabolique de glycanes est une méthode efficace afin de détecter par fluorescence les bactéries Legionella pneumophila à l’aide d’analogues d’un précurseur de l’acide légionaminique et les cellules eucaryotes PC3 via des dérivés d’autres monosaccharidiques. Le marquage obtenu dans les deux cas est membranaire. Cette méthode permet éventuellement de combiner des aspects d’imagerie ou de diagnostic, avec différentes approches thérapeutiquesThis PhD work focuses on glycans metabolic labeling. This method uses a modified monosaccharides bearing a chemical reporter. The unnatural monosaccharide is metabolically incorporated into glycans. Two targets were selected, Legionella pneumophila bacteria and prostate cancer cells (PC3).Various saccharidic analogs were synthesized carrying several chemical reporters, like azide, terminal alkyne, terminal alkene, cyclopropene and ketone functions. A common synthetic strategy was developed using N-hydrosuccinimide ester derivatives and amino monosaccharides, such as D-mannosamine, D-galactosamine and L-fucose for eukaryotic cells and analogs of a legionaminic acid precursor for Legionella pneumophila bacteria.Differents metabolic labeling of glycans were carried out on Legionella pneumophila. Various sugar derivatives were incorporated by bacteria, then the reporter group was reacted selectively and covalently with a complementary function (azide-cyclooctyne, alkene-tetrazine, ketone-hydrazide/alkoxyamine). Bacteria were visualized with an imaging probes by light microscopy (directly: partner bearing a fluorophore, indirectly: recognition of a biotin group by a fluorescent streptavidin). The results highlighted outer membrane labeling.For eukaryotic cells (PC3), metabolic oligosaccharides engineering has been accomplished to check chemical reporter analogs incorporation via detection with fluorescent probes. A series of tools carrying functions complementary to the chemical reporters was synthesized, such as cyclooctyne derivatives (TMDIBO) and tetrazine derivatives. These derivatives were combined with biotin groups for detecting tools or with Antibody Recruiting Molecules (ARMs), such as 2,4-dinitrophenyl and L-rhamnose, for potential therapeutic approaches. The concept focuses on the combination of metabolic glycan labeling and the activation of human serum complement by ARMs to kill selectively labeled cells. First, ARMs labeled cells was checked by recognition of fluorescent anti-ARMs antibodies. Then, metabolic glycan labeling and human serum complement activation has been evaluated but the complement activation protocols still need to be optimized.Metabolic labeling of glycans is an effective method to detect, by fluorescence, Legionella pneumophila bacteria using analogs of a legionaminic acid precursor and eukaryotic cells (PC3) with monosaccharide derivatives. The labeling was observed on the membrane of the two targets. This method can potentially combine imaging or diagnostic aspects with various therapeutic approaches
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Carlier, Mathieu. "Synthèse d’outils pour le marquage métabolique des glycanes." Thesis, Université Paris-Saclay (ComUE), 2019. http://www.theses.fr/2019SACLS453.
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Les glycanes sont des biomolécules constituées d’un enchainement de monosaccharides liés entre eux par des liaisons glycosidiques. La nature et l’abondance des monosaccharides constituant la chaine glycanique ainsi que l’agencement des motifs de glycosylation diffèrent fortement en fonction de l’organisme d’origine. La biosynthèse et la dégradation de ces architectures polysaccharidiques sont finement régulées par des systèmes enzymatiques spécifiques et sont organisées au sein des divers compartiments cellulaires. Les glycanes interviennent dans divers processus biologiques : réserves énergétiques, repliement et stabilité protéique, reconnaissance cellulaire et adhésion à la matrice extracellulaire.Cette thèse porte sur la synthèse et l’utilisation de composés permettant le marquage métabolique des glycanes de cellules eucaryotes (lignée cellulaire du cancer de la prostate, PC-3) ou de bactéries didermes à mycomembranes (Corynebacterium glutamicum).Les composés synthétisés ou utilisés dans cette thèse sont des saccharides fonctionnalisés par un groupement bio-orthogonal (groupements azido, alcyne ou méthylcyclopropène) ou des outils de marquage porteurs simultanément d’un groupement bio-orthogonal complémentaire (groupements cyclooctyne ou tétrazine) et d’une étiquette pouvant être détectée par une macromolécule fonctionnalisée par des fluorophores (D-biotine/stréptavidine ou 2,4-DiNitroPhénol/ anticorps anti-DNP). La solubilité, en milieu aqueux, des outils de marquage est un facteur limitant leur utilisation. Une partie des travaux de cette thèse a consisté à développer des outils de marquage plus solubles en milieu aqueux, par l’ajout d’un motif -sulfo--alanine. Ce motif porte une fonction acide sulfonique qui est déprotonée, à pH physiologique, favorisant ainsi la solubilité en milieu aqueux.L’incorporation métabolique des saccharides fonctionnalisés ainsi que la capacité des outils à marquer les cellules (après incorporation métabolique) est évaluée, sur l’un ou l’autre des modèles biologiques, par analyse en cytométrie en flux ou par microscopie confocaleGlycans are biomolecules made up of a chain of monosaccharides bound together by glycosidic bonds. The nature and abundance ofmonosaccharides forming the glycanic chain and the arrangement of glycosylation patterns differ greatly depending on the organism of origin. The biosynthesis and degradation of these polysaccharide architectures are finely regulated by specific enzymatic systems and are organized within the various cell compartments. Glycans are used in various biological processes: energy reserves, protein folding and stability, cell recognition and adhesion to the extracellular matrix.This thesis focuses on the synthesis and use of compounds allowing the metabolic labeling of glycans of eukaryotic cells (cell line of prostate cancer, PC-3) or of diderm bacteria with mycomembranes (Corynebacterium glutamicum).The compounds synthesized or used in this thesis are saccharides functionalized by a bio-orthogonal groups (azido, alkyne or methylcyclopropene groups) or labeling tools simultaneously carrying a bio-orthogonal complementary function (cyclooctyne or tetrazine groups) and a label that can be detected by a macromolecule functionalized by fluorophores (D-biotin/streptavidine or 2,4-DiNitroPhenol/ anti-DNP antibodies). The solubility of labeling tools in aqueous media is a limiting factor. Part of the work of this thesis has been to develop more water-soluble labeling tools by adding a -sulfo--alanine pattern. This pattern has a sulfonic acid function, which is deprotonated at physiological pH, thus promoting aqueous solubility.The metabolic uptake of functionalized saccharides and the ability of tools to label cells (after metabolic uptake) are evaluated in either biological model, by flow cytometry analysis or confocal microscopy
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Adua, Eric. "N-Glycosylation profiles as a risk stratification biomarker for Type II Diabetes Mellitus and its associated factors." Thesis, Edith Cowan University, Research Online, Perth, Western Australia, 2018. https://ro.ecu.edu.au/theses/2162.
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Worldwide, the prevalence of cardiometabolic diseases, particularly type II diabetes mellitus (T2DM), and to a lesser extent, metabolic syndrome (MetS), has increased dramatically. Despite this increase, there is still a lack of robust biomarkers for cardiometabolic diseases to secure better clinical outcomes. The enzymatic attachment of oligosaccharides (glycans) to proteins-glycosylation is of metabolic and physiological significance, as exploring aberrations of glycosylation profiles can reveal novel biomarkers. In parallel, this process could also explain the biological mechanisms that underpin a suboptimal health status (SHS), a reversible subclinical stage of a cardiometabolic disease. However, studies on the correlation between glycosylation and MetS/T2DM are scarce and none has thus far been performed on a West African population. Thus, the overall aim of this thesis was to explore complementary biomarker panels of healthy and diseased patients considered relevant to Ghanaian residents.
The thesis is structured in the form of five related studies, each addressing a specific aim. From January 2016 to October 2016, a longitudinal case-control study comprising 253 T2DM patients and 260 controls, aged 18-80 years was conducted in Ghana. Fasting plasma samples were collected for clinical assessment, after which plasma N-glycans were analysed by Ultra-Performance Liquid Chromatography (UPLC) and statistical analyses performed. Central adiposity, underweight, high systolic blood pressure (SBP), high diastolic blood pressure (DBP) and high triglycerides (TG) were found to be independent risk factors associated with high SHS after adjusting for age and gender (Study I). SHS score was associated with age, physical inactivity, fasting plasma glucose (FPG), TG and MetS. MetS was associated with increased high branching (HB), trigalactosylated (G3), antennary fucosylated (FUC_A), triantennary (TRIA) and decreased low branching (LB) glycan structures (Study II). The levels of HB, G3, FUC_A, and TRIA N-glycans were increased in T2DM whereas levels of LB, non-sialylated (S0), monogalactosylation (G1), core fucosylation (FUC_C), biantennary galactosylation (A2G) and biantennary (BA) Nglycans were decreased compared to controls (Study III). Biguanides alone, or in combination with sulfonylurea and thiazolidinedione, did not improve glycaemic status at follow-up. Many participants using angiotensin converting enzyme inhibitors achieved desired targets for blood pressure control while statins were effective for control of plasma lipids (Study IV). At a population level, the variability of N-glycan structures ranged from 11% to 56% at both baseline and follow-up, with an average coefficient of variation of 28% and 29%, respectively. The intra-individual N-glycan peak (GP) variations were minor except for GP1 and GP29. However, there were no statistically significant differences in N-glycosylation profiles from baseline to follow-up (Study V).
This thesis shows an association between SHS and MetS/T2DM while MetS and T2DM are characterised by increased levels of complex N-glycan structures, and these structures are stable in T2DM over six months. Many of the findings in this thesis agree with earlier studies from Chinese and Croatian populations with major differences attributed to genetic and environmental factors. Future longitudinal studies are required to provide a better understanding of the transition from SHS to T2DM, as well as to validate N-glycans as generic risk stratification biomarkers for a general population.
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Peroni, Odile. "Mesure de la néoglucogénèse chez le rat à l'aide de [2-13C] glycérol et de l'analyse de la distribution des isotopomères de masse du glucose : étude in vitro (foies isolés perfusés) chez l'animal sain et in vivo chez l'animal sain ou présentant un diabète insulinopénique induit par la streptozotocine." Lyon 1, 1997. http://www.theses.fr/1997LYO1T321.
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Goßner, Sophia Karolina [Verfasser], Karl-Heinz [Akademischer Betreuer] Engel, Karl-Heinz [Gutachter] Engel, and Michael [Gutachter] Rychlik. "Impact of Cross-Breeding on the Contents of Inositol Phosphate Isomers and the Metabolite Profiles of Low Phytic Acid Soybean (Glycine max (L.) Merr.) Mutants / Sophia Karolina Goßner ; Gutachter: Karl-Heinz Engel, Michael Rychlik ; Betreuer: Karl-Heinz Engel." München : Universitätsbibliothek der TU München, 2019. http://d-nb.info/1192911601/34.
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Wargnies, Marion. "Adaptations métaboliques de Trypanosoma brucei en réponse à des variations des conditions intra- et extracellulaires." Thesis, Bordeaux, 2016. http://www.theses.fr/2016BORD0200/document.
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Trypanosoma brucei est un parasite protozoaire responsable de la trypanosomiase humaine africaine. Il présente un cycle de vie complexe alternant entre des hôtes mammifères et un vecteur insecte, la mouche tsé-tsé. Au cours de ce cycle, il rencontre des environnements radicalement distincts auxquels il s’adapte en régulant son métabolisme. Nous avons étudié le métabolisme intermédiaire et énergétique de la forme procyclique évoluant dans le tractus digestif de l’insecte vecteur. Dans cet environnement dépourvu de glucose, la néoglucogenèse est cruciale pour la croissance et la survie des parasites car elle permet la synthèse d’hexoses phosphates et en particulier du glucose 6-phosphate qui alimente plusieurs voies de biosynthèse essentielles. Nos travaux confirment ce flux néoglucogénique alimenté par la proline mais aussi par le glycérol. Nous montrons que le glycérol est une source de carbone efficacement métabolisée et préférentiellement utilisée par la forme procyclique à défaut de la proline et même du glucose pour alimenter son métabolisme intermédiaire. Cette situation qu in’a jamais été décrite auparavant met en évidence la répression du glycérol sur le métabolisme du glucose. Nous montrons également que l’enzyme fructose 1,6-biphosphatase(FBPase), spécifique de la néoglucogenèse, n’est pas essentielle à la survie du parasite en conditions dépourvues de glucose indiquant qu’il existe une alternative à cette enzyme.Toutefois, FBPase joue un rôle important dans la virulence de T. brucei dans l’insecte.De plus, nous avons mis en évidence une autre stratégie d’adaptation de T. brucei basée sur des réarrangements génomiques qui peuvent mener à la synthèse de gènes chimèresTrypanosoma brucei is a protozoan parasite responsible for human African trypanosomiasis. His complex life cycle alternates between mammalian hosts and the insect vector, the tsetsefly. During this cycle, the parasite encounters dissimilar environments and adapts to the sechanging conditions by regulating his metabolism. We have studied intermediate and energetic metabolism of the procyclic form living in the midgut of the insect vector. In this glucose-depleted environment, gluconeogenesis is crucial for growth and viability of the parasites. Indeed, it allows the synthesis of hexoses phosphates and in particular glucose 6-phosphate which feeds several essential biosynthetic pathways. Our work has confirmed the existence of a gluconeogenic flux fed by proline and glycerol. We have shown that glycerol is an efficiently metabolized carbon source and is preferentially used by the procyclic form rather than proline or even glucose. This situation never described before highlights glycerol repression on glucose metabolism. We have also showed that the enzyme fructose 1,6-biphosphatase (FBPase), specific of the gluconeogenesis, is not essential for the viability ofthe parasite in glucose-depleted conditions, suggesting that there is an alternative to this enzyme. However, FBPase plays an important role for virulence of T. brucei in the insect. Moreover, we have showed another adaptation strategy developed by T. brucei which is basedo n genomic rearrangements leading to the synthesis of chimeric genes
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Claret, Carole. "Métabolismes oxydatif et fermentaire du glycérol chez les bactéries : étude physiologique et cinétique de sa conversion en dihydroxyacétone et en 1,3-propanediol." Toulouse, INSA, 1992. http://www.theses.fr/1992ISAT0034.
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L'etude cinetique et physiologique de la conversion du glycerol en dihydroxyacetone par gluconobacter oxydans montre que cette fermentation est soumise a une double inhibition. L'effet inhibiteur du substrat se manifeste par une diminution des cinetiques de croissance et de production avec l'augmentation de la teneur en glycerol, modelisee par des lois de type exponentiel. Cet effet est essentiellement lie a l'abaissement de l'activite de l'eau par les concentrations elevees en substrat. L'inhibition par la dha se manifeste tout d'abord par une diminution des cinetiques de croissance et d'oxydation du substrat (faibles concentrations) puis par un blocage du developpement cellulaire et de la synthese de dha (concentrations elevees). Ces phenomenes, modelises par des lois de levenspiel, ont ete attribues a la forte reactivite de la dha avec les enzymes cytoplasmiques et impliques dans le transport du glycerol et avec la glycerol deshydrogenase membranaire, responsable de la formation de dha. Un procede bi-phasique mono-etage, integrant ces contraintes biologiques, a alors ete mis au point, permettant d'ameliorer les performances fermentaires de cette transformation. Le metabolisme fermentaire du glycerol a ete aborde sous un angle microbiologique. Deux micro-organismes responsables de la degradation du glycerol au sein d'une flore microbienne anaerobie ont ete isoles et caracterises: clostridium butyricum cncm 1. 1211, qui transforme le glycerol en 1,3-propanediol et butyrate avec des rendements de conversion respectifs de 0,66 et 0,13; enterobacter agglomerans cncm 1. 1210, espece nouvellement decrite pour ses capacites d'assimilation du glycerol et de conversion de ce substrat en acetate et 1,3-propanediol (rendements respectifs de 0,25 et 0,7). La preponderance du role joue par le systeme accepteur final d'equivalents reducteurs sur la regulation du metabolisme fermentaire quelle que soit l'espece microbienne etudiee a egalement ete definie
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Bataille, Alain R. "Genome wide search for glycine metabolism genes." Thesis, 2003. http://spectrum.library.concordia.ca/2260/1/MQ83846.pdf.
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The yeast Saccharomyces cerevisiae uses glycine for the synthesis of activated one-carbon units and nitrogen metabolism. The homozygous diploid set of deletion strains includes strains deleted from all the S. cerevisiae non-essential genes (about 4700). This deletion set was screened for strains exhibiting a growth phenotype when glycine was used as the sole nitrogen source (Gmin medium). This screen identified a total of 321 strains. Within the 321 strains, 250 showed a growth defect on Gmin (G- phenotype) whereas 71 exhibited better growth on Gmin (G+ phenotype). Further high-density colony-array analyses established the growth profile for each of the 321 strains on ten different media. These growth profiling experiments identified genes important for various aspects of glycine metabolism. For example vesicular transport from the plasma membrane to the multivesicular body compartment was important for growth in media containing high concentrations of glycine. This suggests that the general amino acid permease (Gap1p) must be targeted to the vacuole for degradation to prevent glycine toxicity. Growth profiling also confirmed the importance of the mitochondrial compartment for glycine metabolism and defined the functional role of CEM1 and YJL046W for lipoate metabolism. This study therefore validated the use of growth profiling data generated using high density colony assays for the study of eukaryotic gene function.
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Riché, Raphaëlle. "Zebrafish model of Glycine Encephalopathy." Thèse, 2018. http://hdl.handle.net/1866/21390.
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Vairinhos, Franklin. "Nitrogen metabolism in cultured `Rhizobium` and in modules of `Glycine max` / by Franklin Vairinhos." 1986. http://hdl.handle.net/2440/21444.
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Bibliography: leaves 189-210xxxiv, 210 leaves : ill ; 30 cm.
Title page, contents and abstract only. The complete thesis in print form is available from the University Library.
Thesis (Ph.D.)--University of Adelaide, Dept. of Agricultural Biochemistry, 1987
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Vairinhos, Franklin. "Nitrogen metabolism in cultured `Rhizobium` and in modules of `Glycine max` / by Franklin Vairinhos." Thesis, 1986. http://hdl.handle.net/2440/21444.
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Liou, Jin-Shuenn, and 劉金順. "Influence of Carbon and Nitrogen Sources on Carbon-Nitrogen Metabolism for Soybean [Glycine max (L.) Merr.] Callus." Thesis, 1995. http://ndltd.ncl.edu.tw/handle/19127680361302003480.
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碩士國立中興大學
植物學系
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Three kinds of callus induced from soybean leaf, hypocotyl and root, were subjected to different carbon and nitrogen sources to investigate the effect on carbon-nitrogen metabolism. The results are summarized as follows: The net photosynthetic rate of the three kinds of callus were negative under all the different carbon and nitrogen sources. The net photosynthetic rate was decreased under exogenous carbon supply. The dark respiratory rate was increased under exogenous carbon supply, while it was not so obvious under exogenous nitrogen supply. It indicated that carbon source supplied the substrates for carbon matebolism and the increase of respiration contributed to the uptake and metabolism of supplied carbon. The accumulation of soluble sugar and total non-structural carbohydrate in three kinds of callus. Carbon source was more obvious under than nitrogen sources and carbohydrate content increase in accord with the amount of carbon source fertilized. Additionally, the accumulation of carbohydarte in leaf callus caused the feedback inhibition in sucrose phosphate synthase activity, but both of hypocotyl callus and root callus were not so similarly inhibited as leaf callus. The soluble protein content increased in accord with the amount of carbon source fertilized, because the carbon source supplied the carbon-skeleton for protein biosynthesizing. But the enzyme activities of nitrogen metabolism in three kinds of callus had different effects under carbon and nitrogen sources treatment. It showed that the ability of nitrate and ammonium utilization was not similar in three kinds of callus.
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Silva, Liliana Adelina Gonçalves da. "Conjugation of fatty acids with coenzyme A, carnitine or glycine : role in mitochondrial energy metabolism and detoxification." Master's thesis, 2013. http://hdl.handle.net/10451/11552.
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Tese de mestrado, Ciências Biofarmacêuticas, Universidade de Lisboa, Faculdade de Farmácia, 2013Carnitine and glycine conjugation are two distinct phase II reactions with relevance for detoxification, and share common substrates, the acyl-Coenzyme A (CoA) esters. The metabolism of valproic acid (VPA), an “epigenetic” drug in clinical practice for decades, has been used within our group as a model system to elucidate drug-induced mitochondrial dysfunction and liver disease, probably common to other carboxylic acid xenobiotics which may be activated to CoA esters. The pharmacotoxicological properties of VPA and their unequivocal interaction with mitochondrial energy metabolism are at the basis of the current project, whose prime objectives were: 1.) the characterization in biological samples of metabolites biomarkers of mitochondrial energy metabolism dysfunction; 2.) application of high resolution chromatographic methods coupled to mass spectrometry to the study of metabolites conjugates, namely the glycine and carnitine esters (acylglycines and acylcarnitines, respectively). Both studies may provide information on the homeostasis of the precursors, acyl-CoAs, and ultimately, the fatty acids and organic acids. A thorough approach on the metabolic significance of acylglycines in biological fluids was undertaken, using stable isotope dilution gas chromatography coupled to mass spectrometry to measure propionyl-, butyryl-, isobutyryl-, 2-methylbutyryl-, isovaleryl-, 3-methylcrotonyl-, hexanoyl- and suberylglicine. The in vivo profiles of these intermediary metabolites were evaluated in biological samples from Wistar rats subjected to a VPA regimen; pediatric patients undergoing chronic treatment with VPA and patients identified with inborn errors of metabolism. The accumulation of acyl-CoA esters intermediates gave rise to a VPA-associated increase on the corresponding products, acylglycines. In addition, novel insights on VPA biotransformation were achieved. As predicted, propionyl-CoA, the end product of VPA thiolytic cleavage was converted to propionylglycine adding a novel phase II reaction to VPA metabolism. Moreover, we initiated the set-up of a new method in our laboratory for the qualitative characterization of carnitine and acylcarnitines using positive electrospray tandem mass spectrometry in plasma and urine. The characterization of short- to long-chain acylcarnitines profiles was performed and additional studies are in course to validate the procedure and quantify the compounds. The differential determination of free carnitine and acylcarnitines levels including acetylcarnitine or palmitoylcarnitine, will support the study of fatty acid oxidation flux, the functional analysis of carnitine acyltransferases activity or the biomonitoring of carnitine or acetylcarnitine as potential therapeutic agents. The overall obtained results allowed to study not only the intermediary metabolism in vivo, which reflects the function and/or dysfunction in cell, but also the biochemical pathways of VPA detoxification, potentially common to other acidic drugs, their metabolites or endogenous molecules.