La thèse a soulevé deux défis dans l’analyse des métaux dans le riz : (i) un dosage multiélémentaire dans le but de la classification du riz selon l’origine géographique, et (ii) la spéciation du sélénium, élément essentiel dans la nutrition, dans le but de l’identification des cibles moléculaires de son assimilation et stockage. Une méthode d’analyse multiélémentaire par l’ICP MS combinée avec des techniques chimiométriques a été proposée comme outil pour la discrimination des riz selon leurs origines géographiques. Les concentrations des 21 éléments ont été analysées par le diagramme en radar, l’analyse en composantes principales, et l’analyse discriminante linéaire. Le riz thaï (riz jasmin) a pu être discriminé des riz étrangers. De plus, l’analyse discriminante a permis de de différencier les riz produits dans les différentes régions de Thaïlande. L’approche de protéomique assistée par l’ICP MS a permis l’identification d’une douzaine de protéines accumulant préférentiellement le sélénium sous forme de sélénomethionine et de sélénocystéine (en proportion 2:1). / The PhD work is focused on two major analytical chemistry challenges concerning rice characterization: (i) the geographical origin classification of rice based on multi-element fingerprinting and (ii) speciation of selenium with the goal of the identification of molecular target of it assimilation and storage in rice. The methods based on the multi-element fingerprinting by inductively coupled plasma mass spectrometry (ICP-MS) in combination with multivariate statistical techniques were developed and validated as tools for authentication of rice. Twenty-one key variables were assessed by a radar plot technique and multivariate data analysis, including principal component analysis (PCA) and discriminant analysis (DA) enabling classification according to geographical origin. Thai jasmine rice could be clearly differentiated from foreign rice samples. In addition, the DA could be used to classify Thai jasmine rice obtained from different regions in Thailand. In the second part of the project, an analytical ICP-MS-assisted proteomic method was developed for the identification of Se-containing proteins in rice. Selenium was found to be present as both selenomethionine (SeMet) and selenocysteine (SeCys) in a dozen of rice proteins with the Se/S substitution ratio two times higher for SeMet than that for SeCys.

Submitted by Marlucy Farias Medeiros (marlucy.farias@unipampa.edu.br) on 2018-02-06T14:51:48Z No. of bitstreams: 1 EDUARDO CESAR PACHLA - 2017.pdf: 13664238 bytes, checksum: 1ee80147db8a398f429417a34a366ed3 (MD5) / Approved for entry into archive by Marlucy Farias Medeiros (marlucy.farias@unipampa.edu.br) on 2018-02-06T15:33:47Z (GMT) No. of bitstreams: 1 EDUARDO CESAR PACHLA - 2017.pdf: 13664238 bytes, checksum: 1ee80147db8a398f429417a34a366ed3 (MD5) / Made available in DSpace on 2018-02-06T15:33:47Z (GMT). No. of bitstreams: 1 EDUARDO CESAR PACHLA - 2017.pdf: 13664238 bytes, checksum: 1ee80147db8a398f429417a34a366ed3 (MD5) Previous issue date: 2017-12-21 / Visando desenvolver produtos com elevado desempenho térmico-acústico, foram produzidos compósitos introduzindo-se resíduos da indústria orizícola em uma matriz de concreto celular. O intuito do trabalho foi averiguar o efeito da adição desses subprodutos à matriz. No total, foram analisadas dez misturas, uma delas de concreto celular, contendo apenas casca do arroz, e as demais, sendo combinações da casca com a palha do arroz em três diferentes comprimentos e percentuais. Assim, o compósito com melhor desempenho foi estudado quanto à adição das fibras saturadas. Todos os compósitos foram avaliados quanto ao desempenho mecânico, acústico e térmico. A partir das análises de resistência mecânica, verificou-se que a adição de casca do arroz reduziu a resistência à compressão axial e aumentou a resistência à tração na flexão. Contanto, a substituição de parcelas de casca por palha do arroz manteve os resultados de resistência à compressão axial estatisticamente semelhantes até o volume de 10%, porém, a partir de 15% de adição de palha, os mesmos foram reduzidos. No que diz respeito à resistência à tração na flexão, a adição de palha do arroz foi positiva. Em relação ao desempenho acústico, a absorção acústica apresentou um acréscimo de aproximadamente 72% com a adição da casca do arroz, e de 83% com a adição de 15% de palha de 3 cm de comprimento. Os resultados de isolamento acústico não se mostraram estatisticamente diferentes, portanto, não foi possível determinar se as fibras vegetais trouxeram ganho na capacidade de isolação sonora. No que concerne o desempenho térmico, constatouse que há redução da condutividade em função do aumento do comprimento da palha do arroz. Por fim, as análises de durabilidade indicaram que os materiais possuem grandes quantidades de hidróxido de cálcio, que por sua vez, em contato com o dióxido de carbono, formam carbonato de cálcio, o que acarreta no surgimento de manchas brancas à superfície dos produtos e acelera o processo de despassivação de reforços metálicos. A concepção dos produtos se mostrou promissora ao obter desempenho térmico-acústico satisfatório, entretanto, é necessário realizar estudos mais minuciosos em relação à durabilidade, e, como alternativa para solucionar essa questão, sugere-se adicionar pozolanas para controlar a formação de hidróxido de cálcio. / Aiming to develop products with high thermal-acoustic performance, composites were produced by introducing waste from the rice industry into a cellular concrete matrix. The purpose of this work was to investigate the effect of adding these byproducts to the matrix. In the total, ten mixtures were analyzed, one of them of cellular concrete, containing only rice husk, and the others, being combinations of the bark and the rice straw in three different lengths and percentages. Thus, the composite with the best performance was studied in the light of addition of saturated fibers. All composites were evaluated for mechanical, acoustic and thermal performance. From the mechanical strength analysis, it was verified that the addition of rice husk reduced the axial compressive strength and increased the tensile strength in the bending. As a result, the substitution of rind plots per rice straw maintained the statistically similar axial compression resistance results up to the 10% volume, but, from 15% straw addition, those results were reduced. Regarding the flexural tensile strength, the addition of rice straw was positive. Considering the acoustic performance, the acoustic absorption presented an increase of approximately 72% with the addition of the rice husk, and 83% with the addition of 15% of straw of 3 cm in length. The results of acoustic insulation were not statistically different; therefore, it was not possible to determine if the vegetal fibers brought positive results to the sound insulation capacity. In terms of thermal performance, it was verified that there is reduction of the conductivity due to the increase of the length of the rice straw. Finally, the durability analysis indicated that the materials have a large amount of the calcium hydroxide, which in turn, in contact with carbon dioxide, forms calcium carbonate, which causes white spots to appear on the surface of the products and accelerates the process of depassivation of metallic reinforcements. The design of the products has shown to be promising due to its satisfactory thermal-acoustic performance, however, it is necessary to carry out more detailed studies on durability and, as an alternative to solve this question, it is suggested to add pozolanas to control the formation of calcium hydroxide.

Shuk-man Chow. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2004. / Includes bibliographical references (leaves 171-186). / Abstracts in English and Chinese. / Thesis committee --- p.i / Statement --- p.ii / Abstract --- p.iii / Acknowledgement --- p.viii / General abbreviations --- p.x / Abbreviations of chemicals --- p.xi / List of figures --- p.xii / List of Tables --- p.xiii / Table of contents --- p.xv / Chapter 1. --- Literature review / Chapter 1.1. --- General introduction to rice disease --- p.1 / Chapter 1.1.1. --- Pathogenesis of Bacterial Leaf Blight (BLB) --- p.1 / Chapter 1.1.2. --- Pathogenesis of rice blast --- p.2 / Chapter 1.1.3. --- Control of rice diseases --- p.3 / Chapter 1.2. --- Plant defense mechanisms --- p.4 / Chapter 1.2.1. --- Basal resistance in plants --- p.4 / Chapter 1.2.2. --- Wound induced defense response --- p.5 / Chapter 1.2.3. --- Pathogen induced host defense response --- p.6 / Chapter 1.3. --- Structure of R gene products --- p.7 / Chapter 1.4. --- Recognition between R and Avr proteins in rice --- p.8 / Chapter 1.5 --- Current knowledge on Xa resistance and AvrXa avirulence protein --- p.9 / Chapter 1.6 --- Current knowledge on Pi resistance and AvrPi avirulence protein --- p.10 / Chapter 1.7 --- Pathogen induced signal transduction cascade --- p.12 / Chapter 1.7.1. --- R gene mediated signal transduction cascade --- p.12 / Chapter 1.7.2. --- Signal events of G-protein activation --- p.12 / Chapter 1.7.3. --- Signaling events for the accumulation of Ca2+ in cytosol --- p.13 / Chapter 1.7.4. --- Signaling events for oxidative burst --- p.14 / Chapter 1.7.5. --- MAPK cascade in defense signaling --- p.15 / Chapter 1.7.6. --- Transcriptional regulation of disease resistance related genes --- p.16 / Chapter 1.7.7. --- Translational regulation of disease resistance related genes --- p.17 / Chapter 1.8. --- Defense responses and defense related genes --- p.19 / Chapter 1.8.1. --- Pathogenesis related (PR) proteins --- p.20 / Chapter 1.8.2. --- Phytoalexins --- p.21 / Chapter 1.9. --- Disease resistance related genes common between rice blast and BLB resistance --- p.22 / Chapter 1.10. --- SA induced signal transduction pathway in rice --- p.23 / Chapter 1.11. --- Important tools facilitating the identification of disease resistance related genes from BLB resistant rice lines --- p.24 / Chapter 1.12. --- Hypothesis --- p.26 / Chapter 1.13. --- Project objective --- p.26 / Chapter 2. --- Materials and Methods --- p.27 / Chapter 2.1. --- Plant Materials --- p.27 / Chapter 2.2. --- Pathogen Inoculation --- p.27 / Chapter 2.3. --- RNA extraction --- p.29 / Chapter 2.4. --- Denaturing gel electrophoresis --- p.29 / Chapter 2.5. --- Subtraction libraries construction --- p.30 / Chapter 2.5.1. --- Cloning of disease resistance related genes --- p.32 / Chapter 2.5.1.1. --- pBluescript II KS (+) T-vector preparation --- p.32 / Chapter 2.5.1.2. --- Ligation --- p.32 / Chapter 2.5.1.3. --- Transformation --- p.32 / Chapter 2.5.1.4. --- Colony picking --- p.33 / Chapter 2.5.1.5. --- PCR amplification of DNA inserts --- p.33 / Chapter 2.5.1.6. --- Purification of PCR products --- p.34 / Chapter 2.6. --- Gene chips printing --- p.34 / Chapter 2.7. --- Probes synthesis and gene chips hybridization --- p.35 / Chapter 2.8. --- Standard-RNAs synthesis --- p.35 / Chapter 2.9. --- Data collection and analysis --- p.36 / Chapter 2.10. --- Sequencing --- p.36 / Chapter 2.11. --- cDNA synthesis --- p.37 / Chapter 2.12. --- RT-PCR --- p.38 / Chapter 2.13. --- DNA gel electrophoresis --- p.39 / Chapter 3. --- Results --- p.58 / Chapter 3.1. --- Construction of BLB gene chips --- p.58 / Chapter 3.1.1. --- Preparation of cDNA clones for gene chips construction --- p.58 / Chapter 3.1.2. --- Purification of PCR products on microtiter plate --- p.59 / Chapter 3.1.3. --- Gene chips construction --- p.59 / Chapter 3.1.4. --- DNA immobilization --- p.62 / Chapter 3.1.5. --- Probe synthesis --- p.62 / Chapter 3.1.6. --- Gene chip analysis --- p.65 / Chapter 3.1.6.1. --- Scanning --- p.65 / Chapter 3.1.6.2. --- Data analysis --- p.65 / Chapter 3.2. --- "Identification of disease resistance related genes commonly regulated by Xa2, Xal2 and Xal4 BLB resistance loci" --- p.70 / Chapter 3.2.1. --- "Signal perception, transduction and regulatory elements" --- p.71 / Chapter 3.2.1.1. --- Proteins involved in reversible phosphorylation cascade --- p.71 / Chapter 3.2.1.2. --- Proteins potentiate signal transduction through specific protein-protein interaction --- p.72 / Chapter 3.2.1.3. --- Other signal transduction components --- p.73 / Chapter 3.2.2. --- Transcriptional and translational regulatory elements --- p.74 / Chapter 3.2.2.1. --- Proteins involved in transcriptional regulation --- p.74 / Chapter 3.2.2.2. --- Proteins involved in post-transcriptional regulation --- p.75 / Chapter 3.2.2.3. --- Proteins involved in translational regulation --- p.76 / Chapter 3.2.3. --- "Oxidative burst, stress, apoptotic related genes" --- p.77 / Chapter 3.2.3.1. --- Stress related proteins --- p.77 / Chapter 3.2.3.2. --- Proteins involved in induction of oxidative burst --- p.78 / Chapter 3.2.3.3. --- PR proteins --- p.79 / Chapter 3.2.3.4. --- Proteolysis related proteins --- p.79 / Chapter 3.2.4. --- Cell maintenance and metabolic genes --- p.80 / Chapter 3.2.4.1. --- Antioxidant --- p.80 / Chapter 3.2.4.2. --- Metabolic genes --- p.81 / Chapter 3.2.4.3. --- Molecular chaperone --- p.82 / Chapter 3.2.4.4. --- Cell cycle regulators --- p.82 / Chapter 3.2.4.5. --- Cell wall maintenance --- p.83 / Chapter 3.2.4.6. --- Proteins involved in protein transport --- p.83 / Chapter 3.2.5. --- Unclassified/others --- p.84 / Chapter 3.3. --- Expression analysis of disease resistance related genes --- p.88 / Chapter 4. --- Discussion --- p.141 / Chapter 4.1. --- Differential expression of disease resistance candidates --- p.141 / Chapter 4.2. --- Disease resistance signal transduction components --- p.143 / Chapter 4.2.1. --- Reversible phosphorylation cascade --- p.143 / Chapter 4.2.2. --- Signal transduction potentiated by protein-protein interaction --- p.144 / Chapter 4.3. --- Other signaling molecules --- p.145 / Chapter 4.3.1. --- PRL1-interacting factor G --- p.145 / Chapter 4.3.2. --- Vacuolar-type H+-ATPasen subunit G --- p.146 / Chapter 4.4. --- Regulation of expression of disease resistance candidates --- p.146 / Chapter 4.4.1. --- Transcriptional regulation of disease resistance related genes --- p.146 / Chapter 4.4.1.1. --- G-box binding protein --- p.147 / Chapter 4.4.1.2. --- MYB TF --- p.147 / Chapter 4.4.2. --- Post-transcriptional modification of disease resistance candidates --- p.148 / Chapter 4.4.2.1. --- RNA splicing factor --- p.148 / Chapter 4.4.2.2. --- Glycine rich RNA binding proteins --- p.149 / Chapter 4.4.3. --- Translational regulation of disease resistance related genes --- p.149 / Chapter 4.5. --- Induction of oxidative burst --- p.150 / Chapter 4.6. --- PR proteins --- p.151 / Chapter 4.7. --- Cell maintenance --- p.152 / Chapter 4.7.1. --- Protein folding --- p.152 / Chapter 4.7.2. --- Protein degradation --- p.153 / Chapter 4.7.3. --- ROS scavenging --- p.154 / Chapter 4.7.4. --- Regulation of cell cycle --- p.154 / Chapter 4.8. --- "Confirmation and profiling of disease resistance related candidates commonly regulated in Xa2, Xal2 and Xal4 BLB resistance NILs at different time points" --- p.155 / Chapter 4.8.1. --- Basal resistance related genes --- p.156 / Chapter 4.8.2. --- General disease resistance related genes --- p.161 / Chapter 4.8.3. --- Pathogen responsive genes --- p.164 / Chapter 4.8.4. --- Prediction of novel genes functions --- p.168 / Chapter 4.9. --- Future prospect --- p.169 / Chapter 4.10. --- Conclusion --- p.169 / References --- p.171 / Appendix --- p.187

Made available in DSpace on 2014-06-11T19:22:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2004-01-30Bitstream added on 2014-06-13T20:48:35Z : No. of bitstreams: 1 alvarez_acc_me_botfca.pdf: 229570 bytes, checksum: 9d9bfcdf8a5f1417f76032419f876733 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O silício apresenta grande potencial na agricultura, revelando-se promissor e eficaz do ponto de vista técnico, pelo fato de aumentar a resistência da parede celular, e dessa forma, diminuir a perda de água por transpiração, aumentar a tolerância a pragas e doenças e resistência ao acamamento, produzindo plantas mais eretas, resultando na melhoria da eficiência fotossintética, o que pode refletir em aumento da produtividade. Nesse contexto, o objetivo do trabalho foi avaliar os efeitos promovidos pela aplicação de silício e diferentes doses de nitrogênio em cobertura (0, 30, 60, 90 kg de N ha-1) nos componentes da produção e produtividade da cultura do arroz, em condições de sequeiro e de irrigação por aspersão. Assim, experimento foi desenvolvido no ano agrícola de 2002/03, num LATOSSOLO VERMELHO Distrófico. O delineamento experimental utilizado foi o de blocos casualizados com parcelas subdivididas e quatro repetições. Foram avaliados os seguintes parâmetros: teor de nitrogênio e silício na folha bandeira no florescimento, teor de silício na planta no final do ciclo, quantidade de silício no solo, altura da planta (cm), acamamento, número de panículas e colmos por metro quadrado, número total de espiguetas por panícula, número de espiguetas granadas e chochas por panícula, peso de 100 grãos, fertilidade dos colmos e das espiguetas, produtividade e rendimento de engenho. Os componentes da produção e a produtividade de grãos não foram influenciados pela aplicação de silício no sulco de semeadura. A irrigação por aspersão aumentou a altura da planta, o número de espiguetas granadas por panícula e a produtividade de grãos. A adubação nitrogenada em cobertura aumentou o número de panículas m-2 e o número de espiguetas por panícula, no entanto, reduziu a fertilidade das espiguetas e a massa de 100 grãos. Em razão da maior disponibilidade hídrica proporcionada pelo sistema de cultivo irrigado por aspe / Silicon presents great agricultural potential, revealing promise and efficacy from a technical standpoint by virtue of increased cell-wall resistance, thus diminishing transpiration water loss, enhancing pests and diseases tolerance and also resistance to lodging, consequently producing more erect plants that attain improved photosynthetic efficiency that can be reflected in productivity growth. In this context, the objective of the present work was to evaluate the effects of silicon application and different rates of nitrogen via top-dressing (0, 30, 60, 90 kg of N ha-1) on production components and productivity in upland system and irrigated by using sprinkler irrigation. This experiment was realized in Dystrophic RED LATOSSOLO during the 2002/2003 agricultural season. The experimental delineation utilized was of causal blocks subdivided into four repetitions. The following parameters were evaluated: concentrations of nitrogen and silicon in the leaf blades at the flowering time, silicon concentration in the plant at the end of the cycle, silicon quantity in the soil, plant height (cm), lodging, number of panicles and stems per square meter, total number of spikelets per panicle, number of spikelets grenades and no grenades per panicle, 100-grain weight, fertility of stems and spikelets, productivity and industry surrending. Neither production components nor productivity was influenced by silicon application into seed furrows. Irrigation increased plant height, the number of spikelets grenades per panicle and grain yield. Nitrogen fertilizer in top-dressing increased the number of panicles per m-2 and the number of spikelets per panicle, but reduced the fertility of spikelets and the 100-grain weight. Due to the greater hydric availability realized in the system irrigated by sprinkler, grain productivity increased linearly with nitrogen fertilizer, while highland cultivation upland presented a response up to 37 kg of N ha-1.

Endocytosis is essential for eukaryotic cells. However, relatively little is known about the endocytic pathway and its molecular machinery in plant cells. In this research, a highly conserved membrane protein called secretory carrier membrane protein 1 (SCAMP) from rice (Oryza sativa) (OsSCAMP1) was employed as a tool to study the plant endocytosis. Toward this goal, I have generated polyclonal antibodies specific to SCAMP and transgenic tobacco BY-2 cell lines expressing yellow fluorescence protein (YFP)-SCAMP fusion. Confocal microscopy study showed that SCAMP localized to both plasma membrane (PM) and motile organelles. Further drug treatment and uptake studies demonstrated that these organelles are early endosomes distinct from Golgi and prevacuolar compartment (PVC), because they colocalized with the endosomal marker FM4-64. Immunogold electron microscopy study with SCAMP antibodies has identified the early endosome (EE) as a vesicular tubular membrane organelle, which resembles the structure of trans-Golgi network (TGN). These results indicate that the secretory and endocytic pathways are merged at the TGN which may serve as the sorting station for both pathways. / Since brefeldin A (BFA) induced both TGN and Golgi to form similar aggregates or BFA compartments in tobacco BY-2 cells, studies were also performed to sort out these BFA-induced compartments. Here I have demonstrated that the BFA-induced compartments derived from Golgi and TGN are physically distinct where the TGN aggregates were usually found to be surrounded by the Golgi aggregates in the same cells in both confocal immunofluorescent and immunogold EM studies. Furthermore, the internalized endosomal marker FM4-64 was found to colocalize with the TGN-derived BFA compartments but separated from the Golgi aggregates, whereas the endocytosis inhibitor tyrphostin A23 prevented TGN but not Golgi from forming BFA compartments. Therefore, the secretory Golgi organelle is functinally distinct from the endocytic TGN/EE in their responses to BFA treatment in plant cells. / The possible roles of SCAMPs in cytokinesis were also investigated. In transgenic tobacco BY-2 cells expressing the TGN/EE marker SCAMP-YFP, SCAMPs were found to be concentrated in the developing cell plate together with the internalized endosomal marker FM4-64 under confocal microscopy and this was further confirmed by immunogold electron microscopy studies with SCAMP antibodies. These results have demonstrated that SCAMPs, TGN and endocytosis are all involved in the cell plate formation during cytokinesis in plant cells. / Lam, Sheung Kwan. / Adviser: Jiang Liwen. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3266. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 176-191). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

No presente estudo, desenvolveu-se um método de análise de especiação de As inorgânico em alimentos derivados de arroz, mediante a técnica de geração de hidretos (HG) associada à espectrometria de absorção atômica (AAS). As condições para a geração seletiva dos hidretos de As(III) e As inorgânico, e As total foram HCl 10 mol L-1 e NaBH4 0,1% (m/v), HCl 6 mol L-1 e NaBH4 0,5% (m/v), respectivamente. Vinte amostras de alimentos à base de arroz comercializadas no Rio Grande do Sul foram analisadas, incluindo bolacha, massa, leite em pó, farinha, mistura para bolo e arroz para sushi; também foram analisadas amostras líquidas tais como vinagre, cerveja, vinho e bebidas lácteas. Nessas amostras, o As total foi quantificado após extração alcalina com K2S2O8 2% (m/v) e HNO3 10% (v/v) sob aquecimento em banho-maria a 90 °C, seguido da determinação do As por HG-AAS. O limite de detecção do As total foi 6,8 ng g-1. As espécies de As nas amostras sólidas foram extraídas utilizando-se HNO3 0,28 mol L-1 e aquecimento em banho-maria a 95 °C, sendo que, nessas condições, não ocorre a interconversão das espécies. Para a quantificação do As inorgânico, foi necessária a pré redução do As, com o emprego de solução de KI/ácido ascórbico, em meio ácido. Os limites de detecção do As(III) e As inorgânico foram 5 ng g-1 e 11 ng g-1, respectivamente. As concentrações de As(III) e As inorgânico nas amostras analisadas variaram, respectivamente, de 9,5 ± 0,6 a 71,8 ± 6,1 ng g-1 e 23,7 ± 6,9 a 82,9 ± 4,7 ng g-1. A concentração de As(V) foi obtida por diferença. A concentração de As total variou de 20,3 a 124 ng g-1. As amostras líquidas, para determinação do As total e do As (III) presente, foram simplesmente diluídas com HNO3 0,28 mol L-1, uma vez que testes preliminares mostraram melhor exatidão mediante diluição das amostras, ao invés da extração com K2S2O8 2% (m/v) e HNO3 10% (v/v). Os limites de detecção do As total variaram de 0,34 a 1,70 μg L-1, enquanto que os do As(III) variaram de 0,50 a 2,50 μg L-1. Com o intuito de se avaliar a exatidão do método foram realizados testes de recuperação do analito. Foi também analisada farinha de arroz certificada, sendo as concentrações de As total e As inorgânico concordantes com os valores de referência. / The present study is about chemical speciation analysis of inorganic As (iAs) in rice products using hydride generation-atomic absorption spectrometry (HG-AAS). Selective generation of arsine of As(III) is achieved with 0.1% (m/v) NaBH4 and 10 mol L-1 HCl. Inorganic arsenic (iAs) is pre reduced and determined in the same conditions used for As(III). Samples of rice products such as flour, cookies, cracker, pasta, milk powder, cakes, rice for sushi, dairy beverage, vinegar, wine and beer were analyzed. Total arsenic (tAs) in solid samples was extracted with 2% (m/v) K2S2O8 and 10% (m/v) HNO3 and heating in water bath at 90 °C for 3 h, while arsenic species were extracted with 0.28 mol L-1 HNO3 and heating in water bath at 95 °C for 1.5 h. The limits of detection (LODs) of the method for As(III), iAs and tAs were 5 ng g-1, 11 ng g-1, and 6.8 ng g-1 respectively. Concentrations of As(III), iAs, and tAs in the solid samples ranged from 16.5 ± 7.2 to 71.8 ± 6.1 ng g-1, 26.7 ± 0.3 to 82.9 ± 4.7 ng g-1, and 35.8 ± 0.4 to 136 ± 2 ng g-1, respectively. Liquid samples were simply diluted in 0.28 mol L-1 HNO3. Arsenic (III) and tAs were determined in these samples. The LODs of As(III) and t(As) were in the range of 0.28 to 0.85 g L-1 and 0.5 to 2.5 g L-1, depending on the sample dilution. The concentration of As(III) in liquid samples was up to 11.4 ± 0.4 5 g L-1 while that of tAs ranged from 1.35 ± 0.12 to 24.6 ± 0.5 g L-1.

Xanthomonas oryzae pv. oryzae (Xoo) est l’agent causal de la bactériose vasculaire du riz (BLB), une maladie dévastatrice dans de nombreux pays rizicoles. Chez cette bactérie, les effecteurs de type TAL (Transcription Activator-Like) jouent un rôle majeur dans le pouvoir pathogène. En effet ces effecteurs qui sont secrétés à l’intérieur des cellules eucaryotes hôtes par le système de sécrétion de type III de Xanthomonas agissent comme de véritables facteurs de transcription capables de manipuler le transcriptome de l’hôte via l’induction de gènes spécifiques. L’interaction entre les TALs et le promoteur hôte ciblé est régi selon un code, tel que la région centrale des répétitions des TALs s’associe spécifiquement à sa séquence cible à raison d’une répétition pour un nucléotide. Un TAL peut agir comme une protéine de virulence via l’induction de gènes dits de sensibilité (S) dont l’activation est nécessaire au développement de la maladie, et comme protéine d’avirulence via l’induction d’un gène dit exécuteur (E), dont l’activation promeut les réponses de défense de la plante. L’objectif de cette thèse était d’identifier et de caractériser de nouvelles sources de résistance dépendants des effecteurs de type TAL pour contrôler la BLB. Chez le riz, les gènes de sensibilité à Xoo les mieux caractérisés sont ceux du clade III de la famille SWEET des transporteurs de sucres. L’un d’entre eux, OsSWEET14, est particulièrement important puisqu’il est ciblé par 4 effecteurs TALs différents et issus de souches de Xoo appartenant à des lignées génétiques distinctes et d’origines géographiques différentes. Partant du constat de la convergence pour l’induction de ce gène S majeur, un crible moléculaire du promoteur de OsSWEET14 a été réalisé dans le but d’identifier du polymorphisme pouvant affecter la liaison TAL-ADN et en conséquence entrainer une perte de sensibilité. Ces travaux ont permis l’identification du gène xa41(t) qui confère une forme de résistance récessive et à large spectre contre Xoo. Dans une seconde partie, le criblage phénotypique d’une centaine de variétés de riz résistantes à la souche africaine de Xoo MAI1 a permis d’identifier cinq TALs agissant comme des protéines d’avirulence. C’est le cas des effecteurs Tal2 et Tal9 qui provoquent spécifiquement une réaction de résistance sur la variété de riz IR64 dont le génome a été séquencé. Des analyses de type RNAseq ont été réalisées et ont permis d’identifier une dizaine de gènes exécuteurs E candidats expliquant potentiellement la résistance de IR64 à la souche de Xoo MAI1. Finalement, une troisième stratégie a été menée dans le cadre d’un projet collaboratif, visant à démontrer que PiCO39 conférant la résistance du riz au champignon pathogène Magnaporthe oryzae pouvait aussi contrôler les bactérioses vasculaire et à stries foliaires. Pour ce faire, des TAL artificiels (dTALE) ont été dessinés pour induire spécifiquement l’expression de la construction de M. oryzae AVR1-CO39 dans des riz transgéniques résistant (PiCO39) et sensible (pico39). Nos données montrent que l’induction de AVR1-CO39 par Xoo ou Xoc conduit de manière PiCO39-dépendante à une réduction spectaculaire des symptômes. Ceci démontre que les réactions de défense induites par un gène de résistance à M. oryzae sont également fonctionnelles contre X. oryzae, ouvrant de nouvelles perspectives originales quant aux stratégies de contrôle des bactérioses dues à Xanthomonas. / Bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo) is the most destructive bacterial disease of rice. Xoo pathogenicity critically depends on the TAL (Transcription Activator-Like) effectors. TALs are type three effectors secreted through a type three secretion system into the eukaryotic cell where they act as transcription factors able to manipulate the host transcriptome via the induction of specific genes. DNA-binding specificity involves a unique central repeated region in the TAL effector whereby each repeat directly binds to one single nucleotide. TALs can act as major virulence effector thtough the induction of so-called susceptibility (S) genes that are essential for disease development, or act as avirulence effector by inducing so-called executor (E) resistance genes that promote host defense responses. The goal of this PhD project was to identify and characterize novel TAL-dependent resistance sources to control BLB. In rice, the best characterized S genes are those of the clade III of the sugar transporters SWEET family. The most important is OsSWEET14 as this gene is targeted at unrelated DNA boxes by four TAL effectors, which belong to strains of different lineages and geographic origins. The evolutionary convergence for the induction of SWEET14 reflects its crucial role as major determinant of rice susceptibility to Xoo. A molecular screening of the OsSWEET14 promoter was performed using the natural diversity of wild African rice in order to identify polymorphism that could affect the TAL/DNA binding and thus lead to loss of susceptibility. This work allowed the identification of xa41(t) that confers broad spectrum recessive resistance to Xoo. In a second part of my PhD project, a phenotypic screen for resistance against the Xoo African strain MAI1 of a hundred of rice accessions enabled to identify five TALs. Among them, Tal2 and Tal9 were shown to trigger resistance on the rice variety IR64 the genome of which is fully sequenced. RNAseq analysis identified a small set of resistance E gene candidates underlying potentially IR64 resistance against Xoo strain MAI1. Finally, as a third strategy we aimed within a collaborative project to investigate if PiCO39 that confers resistance of rice towards Magnaporthe oryzae could also control BLB and BLS (Bacterial leaf streak). To that end, Artificial TAL effectors (dTALe) were designed to induce specifically the M. oryzae AVR1-CO39 construct in resistant (PiCO39) and susceptible (piCO39) transgenic backgrounds. We show that the induction of AVR1-CO39 by Xoo or Xoc drastically impairs bacterial colonization in a PiCO39-dependent manner, highlighting the potential of exploiting rice blast or other resistance genes as novel strategies to control rice pathogenic Xanthomonas bacteria.

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2017-03-30T13:51:58Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_marcio_rogerio_pereira_leite.pdf: 1122444 bytes, checksum: bed87ca85ed26067e56cccdaf3f0f3f3 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-04-05T18:56:55Z (GMT) No. of bitstreams: 2 dissertacao_marcio_rogerio_pereira_leite.pdf: 1122444 bytes, checksum: bed87ca85ed26067e56cccdaf3f0f3f3 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-04-05T18:58:32Z (GMT) No. of bitstreams: 2 dissertacao_marcio_rogerio_pereira_leite.pdf: 1122444 bytes, checksum: bed87ca85ed26067e56cccdaf3f0f3f3 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-04-05T18:58:45Z (GMT). No. of bitstreams: 2 dissertacao_marcio_rogerio_pereira_leite.pdf: 1122444 bytes, checksum: bed87ca85ed26067e56cccdaf3f0f3f3 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-06-02 / Sem bolsa / Este trabalho teve como objetivo comparar a qualidade fisiológica, ao longo do armazenamento, de dois tipos de sementes de arroz de sequeiro separadas durante a debulha, pelos agricultores familiares de Araguatins - TO. O experimento foi implantado no Instituto Federal do Tocantins Campus Araguatins, como também foram enviados amostras para serem analisadas no laboratório de análise de sementes da Universidade Federal de Pelotas, no início e no final do período de armazenamento. Foram utilizadas sementes de arroz de sequeiro, cultivar Agulhinha. Os tratamentos constituíram-se da combinação das formas de obtenção das sementes, ou seja, obtidas sob a bancada de debulha, denominada Semente Tipo 1 e sementes que durante o processo de debulha saltaram para longe da bancada (mais de um metro), chamadas de Semente Tipo 2, combinando-se com 6 épocas de avaliação (40, 70, 100, 130, 160 e 190 dias após a colheita), totalizando 12 tratamentos experimentais, com quatro repetições. Avaliou-se germinação; emergência em campo e laboratório; primeira contagem; índice de velocidade de emergência e massa de 1000 sementes. Conclui-se que não houve diferença significativa entres os tratamentos TIPO 1e TIPO 2 em todas as variáveis resposta, exceção daquelas dos 40, 100 e 130 dias após a colheita, na emergência em laboratório. O sistema de seleção de sementes de arroz de sequeiro utilizado pelos agricultores familiares de Araguatins – TO não é eficiente na separação por qualidade fisiológica. / This study Aimed to compare the physiological quality during storage of two upland rice seeds types, separated during threshing, by the family farmers of Araguatins - TO. The experiment was established at the Federal Institute of Tocantins Campus Araguatins. Samples of seeds are sent to be analyzed in Seed Analysis Laboratory of Federal University of Pelotas, at the beginning and end of the storage period. Rice seeds were used, to the cultivar Agulhinha, the treatments consisted of the combination of the ways of obtaining seeds, or obtained under the threshing bench called Type 1 seed and seed during threshing process jumped away from the bench (more than a meter), Seed called Type 2. That was combined with six evaluation periods (40, 70, 100, 130, 160 and 190 days after harvesting), totaling 12 experimental treatments with four replications. To evaluate germination, emergence to field and laboratory, first count, emergence speed index, weight of 1000 seeds. The results showed no significant difference between seeds TYPE 1 and 2 in all response variables, except those of 40, 100 and 130 days after crop emergence in the laboratory. The selection system of upland rice seeds used by farmers to Araguarina - TO is not effective in separation of seeds by physiological quality.

Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2017-06-23T11:46:07Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_alessandra_vollmann.pdf: 719074 bytes, checksum: fc4a031e8a673f1bbfe20e1289928b22 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-06-23T22:08:26Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_alessandra_vollmann.pdf: 719074 bytes, checksum: fc4a031e8a673f1bbfe20e1289928b22 (MD5) / Made available in DSpace on 2017-06-23T22:08:26Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) dissertacao_alessandra_vollmann.pdf: 719074 bytes, checksum: fc4a031e8a673f1bbfe20e1289928b22 (MD5) Previous issue date: 2016-02-03 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / O armazenamento de sementes é uma atividade essencial, que tem por função a manutenção da qualidade da semente para a próxima semeadura. As sementes de arroz (Oryza sativa L.) podem ser armazenadas tanto em sacaria quanto a granel, buscando a preservação da sua qualidade fisiológica, por meio de condições onde os danos sejam mínimos. Assim sendo, este trabalho teve por finalidade avaliar a influência da temperatura durante o armazenamento na qualidade fisiológica de sementes de arroz cv BR-IRGA 417, bem como verificar possíveis alterações na composição química e na atividade de enzimas hidrolíticas. Para isto, sementes de arroz foram submetidas a quatro condições de armazenamento, sendo, condições sem controle de umidade e temperatura (laboratório), temperatura constante de 15 ºC, temperatura de 15ºC, com elevação da temperatura para 25 ºC, durante 24h, a cada sete dias de armazenamento e sementes armazenadas em temperatura de 15ºC, com elevação da temperatura para 25 ºC, durante 24h, a cada 14 dias de armazenamento, mantidas armazenadas por 168 dias e avaliadas em períodos regulares de 56 dias. Os seguintes testes foram realizados: germinação, primeira contagem da germinação, índice de velocidade de germinação, comprimento de parte aérea e raiz, massa seca de parte aérea e raiz, condutividade elétrica, em todos os períodos avaliados, Já os seguintes testes: composição química e determinação da atividade das enzimas α-amilase e fosfatase ácida, bem como parâmetros obtidos no teste de emergência de plântulas, foram realizados antes de iniciar o armazenamento e após 168 dias. Os resultados mostraram que a alternância de temperatura durante o armazenamento ocasiona redução na qualidade fisiológica da semente. Portanto, pode-se concluir que, apesar de manter o potencial germinativo alto, ocorre redução no vigor das sementes de arroz, cv BR-IRGA 417, quando armazenadas em condições inadequadas de temperatura. / Seed storage has become is an essential activity, whose function is to maintain the quality of seed for the next sowing. Rice seeds (Oryza sativa L.) can be stored either in sacks as bulk, seeking to preserve your physiological quality, through conditions where damage seeds are minimal. Therefore, this experiment aimed at evaluate the influence of temperature during storage on the physiological quality of rice sseds, cv BR- IRGA 417, and verify changes in chemical compositions and activity of hydrolytic enzymes. For that, rice seeds were submitted to four storage conditions, without humidity and temperature control (laboratory), contant temperature of 15 ºC, temperature of 15 ºC with rise temperature to 25 ºC during 24 hours, every seven day storage, and temperature of 15 ºC with rise temperature to 25 ºC during 24 hours, every 14 day storage, remain stored for 168 days and evaluated at regular intervals of 56 days. The following testes were performed: germination test, first count of germination, germination speed rate, shoot and root length, dry mass of shoot and root, electrical conductivity, in all periods, already the following tests: chemical composition, and determining the activity of α-amylase and phosphatase acid, as well as parameters obtained in the emergency seedlings test, were carried out before starting storage and after 168 days. The results showed that during storage temperature alternation causes reduction in the seed physiological quality. Therefore, it can be concluded that though maintaining high germination potential occurs reduction in the effect of rice seeds cv BR-IRGA 417, when stored under unsuitable temperature conditions.

Submitted by Gabriela Lopes (gmachadolopesufpel@gmail.com) on 2017-07-11T17:25:56Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇÃO Renan Souza Silva.pdf: 1484537 bytes, checksum: 7a9b959ee069cd859ca0a1bfaf44da09 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2017-07-17T20:31:44Z (GMT) No. of bitstreams: 2 DISSERTAÇÃO Renan Souza Silva.pdf: 1484537 bytes, checksum: 7a9b959ee069cd859ca0a1bfaf44da09 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-07-17T20:31:44Z (GMT). No. of bitstreams: 2 DISSERTAÇÃO Renan Souza Silva.pdf: 1484537 bytes, checksum: 7a9b959ee069cd859ca0a1bfaf44da09 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-04-13 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / O arroz irrigado (Oryza sativa) é uma cultura que alimenta bilhões de pessoas no mundo inteiro, com produtividades que batem recordes a cada safra. Porém, as plantas daninhas ainda representam problemas para o arroz expressar seu potencial produtivo, com destaque para o arroz-daninho, da mesma espécie que a cultivada. Nos últimos anos, com o advento da tecnologia Clearfield® (CL), muitos produtores viram nessa tecnologia a solução dos problemas de controle do arroz-daninho. Contudo, a possibilidade de fluxo gênico do arroz CL para a espécie daninha trouxeram problemas para a tecnologia. Uma nova linhagem de arroz, que se utiliza de outro modo de ação de herbicidas (inibidores da ACCase), desta vez, está prestes a ser lançada no mercado e se oferece como uma nova ferramenta no processo de combate ao arroz-daninho. Aliada a essa ferramenta, existe a possibilidade de aumento no espectro de controle de plantas daninhas do arroz com associação a outros herbicidas. Para isso é necessário conhecer os efeitos dos produtos associados e a seletividade do arroz frente às associações, pois há possibilidade do surgimento de interações sinérgicas ou antagônicas na associação de latifolicidas com herbicidas desse mecanismo de ação. Embora não seja uma prática prevista em lei, a associação de herbicidas é mais uma alternativa para os orizicultores no manejo de plantas daninhas resistentes. Os objetivos deste estudo foram: efetuar uma revisão de literatura acerca do modo de ação dos herbicidas registrados para o arroz e os efeitos da interação ocasionados pela interação de herbicidas (Capítulo I); elucidar as interações decorrentes da associação entre quizalofop e haloxyfop com herbicidas registrados para o arroz (Capítulo II); e estudar os efeitos de duas linhagens de arroz (suscetível e resistente a inibidores da ACCase) sujeito à aplicação de herbicidas previamente antagônicos (Capítulo III). Os resultados obtidos indicam que houve redução da eficiência de controle do arroz com todas as doses avaliadas na maioria das associações de quizalofop e haloxyfop. Houve suposto antagonismo ou efeito aditivo nas associações de quizalofop com todos os produtos em todas as doses em Aeschynomene denticulata. Outros estudos devem ser realizados com o uso de curvas de dose resposta e montagem de isobologramas para a comprovação da suposição da interação. Os resultados evidenciam suposta interação antagônica entre o 2,4-D e o quizalofop e entre o quizalofop e o propanil nos três biótipos estudados. / Irrigated rice (Oryza sativa) is a crop that feeds billions of people worldwide, with record-breaking yields. However, weeds still represent problems for rice to express its productive potential, especially weedy-rice, of the same species as the cultivated one. Last years, with Clearfield® (CL) technology, many producers have taken a technology as a solution of the weedy-rice problems. However, the possibility of gene flow of CL rice to a weed brought problems with a technology. A new strain of rice, which uses another mode of action of herbicides (inhibitors of ACCase), this time, is about to be launched in the market and aims to be a new tool in the process of combating weedyrice. Allied to this tool, there is a point of increase without control of weed control of the rice with the advent and a legalization of the association of herbicides in the country. For this it is necessary to know the effects of the associated products and a selectivity of the rice in front of the associations, since there is a possibility of synergistic or antagonic interactions in the association of latifolicides with herbicides of this mechanism of action. Although not a legal practice, the association of herbicides is one more alternative for the rice producers in the fight against the problems with resistance of weeds. The objectives of this study were: to carry out a literature review about the mode of action of the herbicides registered for rice and the interaction effects caused by the interaction of herbicides (Chapter I); To elucidate the interactions resulting from the association between quizalofop and haloxyfop with herbicides registered for rice (Chapter II); And to study the effects on rice of two strains (susceptible and resistant to ACCase inhibitors) subject to the application of supposedly antagonistic herbicides (Chapter III). The results indicate that rice control efficiency was reduced with all doses evaluated in most associations of quizalofop and haloxyfop. There was supposed antagonism or additive effect on the associations of quizalofop with all products at all doses in Aeschynomene denticulata. Other studies should be performed using dose response curves and isobolograms assembly to prove the assumption of the interaction verified. The results evidenced the supposed antagonistic interaction between 2,4-D and quizalofop and between quizalofop and propanil in the three studied biotypes.