Submitted by Gabriela Lopes (gmachadolopesufpel@gmail.com) on 2018-06-13T17:18:47Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Luis Henrique Konzen.pdf: 646860 bytes, checksum: 511c31d83c08144defe344c571319e35 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-06-14T20:50:11Z (GMT) No. of bitstreams: 2 Luis Henrique Konzen.pdf: 646860 bytes, checksum: 511c31d83c08144defe344c571319e35 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-06-14T20:50:44Z (GMT) No. of bitstreams: 2 Luis Henrique Konzen.pdf: 646860 bytes, checksum: 511c31d83c08144defe344c571319e35 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-06-14T20:50:50Z (GMT). No. of bitstreams: 2 Luis Henrique Konzen.pdf: 646860 bytes, checksum: 511c31d83c08144defe344c571319e35 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-03-02 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / O estabelecimento da lavoura de tabaco se dá através do transplante de mudas, e o passo inicial para promover um estande uniforme de plantas e garantir uma boa produtividade da lavoura de tabaco é a utilização de mudas de boa qualidade, para isso, é necessário a utilização de sementes de altíssima qualidade. Neste sentido, os testes de vigor são muito importantes para obtenção de informações adicionais ao teste de germinação e podem auxiliar na tomada de decisão. Assim, objetivou-se com o presente trabalho determinar metodologias do teste de envelhecimento acelerado para avaliação do vigor em sementes de tabaco. Para o estudo, foram utilizados dez lotes de sementes de tabaco. Inicialmente determinou-se a qualidade inicial das sementes de tabaco através dos testes de germinação, primeira contagem de germinação, índice de velocidade de germinação, emergência de plântulas avaliadas aos 7, 14 e 21 dias após semeadura, índice de velocidade de emergência e envelhecimento acelerado com água conduzido conforme proposto pela AOSA. Após a determinação da qualidade inicial das sementes, estudou se o teste de envelhecimento acelerado nas metodologias: envelhecimento acelerado com água, solução salina saturada (40g de NaCl.100mL-1 de água) e solução salina não saturada (11g de NaCl.100mL-1 de água), submetidas a temperatura de 45 e 41⁰C, por períodos de exposição de 24, 48 e 72 horas. O ensaio foi conduzido em delineamento de blocos ao acaso, as médias obtidas por lote, em cada avaliação, foram comparadas pelo teste de Scott-Knott em nível de probabilidade de 5% e foi realizada análise de correlação linear. De acordo com os resultados obtidos, conclui - se que o teste de envelhecimento acelerado com água conduzido sob temperatura de 45ºC combinado com período de exposição de 24 horas mostra-se adequado para avaliação do vigor de sementes de tabaco. / The establishment of tobacco is performed through seedling transplant, and the initial step to promote a uniform plant stand and ensure a good yield of the tobacco crop is the use of good quality seedlings, for this, the use of seeds of the highest quality is necessary. In this sense, vigor tests are very important to obtain additional information to the standard germination test and can assist in decision-making. Thus, the aim of this work was to determine accelerated aging test methodologies for the evaluation of vigor in tobacco seeds. For the study, ten lots of tobacco seeds were used. The initial quality of the tobacco seeds was determined through the germination test, first germination count, germination speed index, emergence of seedlings at 7, 14 and 21 days after sowing, emergence speed index and the accelerated aging with water conducted as proposed by the AOSA. After the determination of the initial quality of the seeds, the accelerated aging test was carry out in the following methods: accelerated aging with water, saturated saline solution (40g NaCl 100mL-1 water) and unsaturated saline solution (11g NaCl 100mL-1 water), submitted to temperature of 45 and 41°C, for periods of exposure of 24, 48 and 72 hours. The assay was conducted in a randomized block designed, the averages obtained per lot at each evaluation were compared by the Scott-Knott test at a 5% probability level and a linear correlation analysis was performed. According to the results, it is concluded that the accelerated aging test with water conducted under a temperature of 45°C combined with a 24 - hour exposure period is adequate for evaluating the vigor of tobacco seeds.

EEffffeecctt ooff ddrroouugghhtt oonn tthhee mmeettaabboolliissmm ooff ttaabbaaccccoo ppllaannttss (Nicotiana tabacum L.) Diploma thesis - abstract Lucia Miedzińska, Bc. Work Supervisor: Doc. RNDr. Helena Ryšlavá, CSc. Consultant: RNDr. Veronika Doubnerová, PhD. Drought stress is one of the most frequent form of plant stress, which occurs not only under condition of limited water availability, but also under reduced ability to accept water by roots, for example in salted or cold soils. In this thesis the changes in enzyme activities of: NADP-malic enzyme (EC 1.1.1.40; NADP-ME), phosphoenolpyruvate carboxylase (EC 4.1.1.31; PEPC) and pyruvate, phosphate dikinase (EC 2.7.9.1; PPDK) in tobacco plants (Nicotiana tabacum L., cv. W38) after drought were investigated. Enzyme activities in tobacco leaves were significantly increased during 11 days of stress, PEPC 2-fold, PPDK 3,3- fold and NADP-ME 4-fold compared to control plants. The regulation of NADP-ME and PEPC activities were studied on transcriptional level - by the real-time PCR method and on translational level - immunochemically. The expression of NADP-ME protein and transcription of mRNA for chloroplast NADP-ME isoform were increased, but mRNA for cytosolic isoform was not affected. The protein expression of PEPC was slightly increased, transcription of...

Nowadays, the environmental pollution by heavy metals is very serious problem all around the world. Radionuclides, including uranium, are heavy metals that cause both chemical and radioactive pollution. Naturally occurring uranium is not so dangerous for living organisms. Human activities, especially uranium ore mining and use of phosphate fertilizers, have increased its concentration in the environment with consequent contamination of soil, water and air. Compared to other countries, the Czech Republic is relatively rich in deposits of uranium ore. Extensive mining results in large contaminated areas, containing not only uranium but also other heavy metals and xenobiotics that need to be removed from the environment. One way how to decontaminate soils and waters is phytoremediation. This eco-friendly and cost-effective technique exploits the ability of plants to take up, translocate, transform and sequester xenobiotics. In order to provide functional phytoremediation, it is necessary to understand the mechanisms of plant responses to stress caused by xenobiotics. Therefore in my master thesis, I focused on the impact of uranium on physiological processes of uranium-stressed plants, with the emphasis on carbohydrate metabolism and antioxidative defense mechanism. Powered by TCPDF (www.tcpdf.org)

Made available in DSpace on 2014-08-20T13:32:57Z (GMT). No. of bitstreams: 1 dissertacao_gabriel_klafke.pdf: 1619582 bytes, checksum: dcdfb7127ef88b680dc43b41887f2c86 (MD5) Previous issue date: 2010-03-18 / During the past decades, the development of genetically modified plants became a consolidated reality. Taking advantage of the genetic engineering process, it is possible to obtain modified plants to use as bioreactors in the production of tissue or organs expressing antigens which can be easily used as vaccines. The plant-based expression systems as tomato and lettuce, which attend as models for that process, present innumerous advantages such as conservation of eukaryotic machinery, which promote pos-translational modifications, possibility of large-scale production and development of safer and economically more attractive vaccines. Taking use of that strategy, the Swine mycoplasm hyopneumoniae (SMP) disease can be one important and possible target to either be eradicated or controlled. The SMP, caused by fastidious bacterium Mycoplasma hyopneumoniae, is one of the most important respiratory disease in swine breeding, due to its very high prevalence coupled with associated losses all over the whorld, and has in the recombinant DNA technology a viable alternative in the development of more effective and safe vaccines The objective of my work was to genetically manipulate tobacco plants in order to use them as bioreactor in the production of an antigen against PMS. Tobacco leaves and internodes were cultured in different concentrations of BAP and AIA hormones. The best regeneration results for both explants were seen with 1,5mg.L-1 BAP and 0,1mg.L-1 AIA. The selection test with the kanamycin antibiotic appeared to be highly effective, showing a total inhibition of regeneration with 30 mg.L-1 and 100 mg.L-1 for leaves and internodes respectively. The recombinant colonies of A. tumefaciens, containing ltb-r, were co-cultivated with the internodes and leaves from plants germinated in vitro. The next step, the explants were transferred to the selection medium in order to induce the selection of the putatively transformed cells. The genomic DNA from regenerated and putatively transformed plants were extracted and amplified by PCR, where it was detected the presence of a band referent to ltb r1. The analyses of the integration and the transcription of ltb-r1 were carried out by Southern blot and RT-PCR, respectively. In both techniques, it was possible to confirm the presence of one band which corresponds to the expected size of ltb-r1, supporting the integration and expression of the gene. However, with the tests used here, it was not possible to detect with accuracy the recombinant protein / Nas últimas décadas, o desenvolvimento de plantas geneticamente modificadas tornou-se uma realidade consolidada. Nesse sentido, utilizando-se da engenharia genética, é possível obter plantas servindo como biorreatores na produção de tecidos ou orgãos expressando antígenos que podem ser facilmente utilizados como vacina. Os sistemas de expressão em plantas como tomate, alface, tabaco que servem como modelos desse processo, apresentam várias vantagens, entre elas, a conservação da maquinaria eucariótica que promove as modificações póstraducionais das proteínas e ainda a possibilidade de produção em larga escala. Dentro desta estratégia de produção de proteínas, pode-se citar a pneumonia micoplásmica suína (PMS), causada pelo agente Mycoplasma hyopneumoniae, uma das principais doenças de suínos que provoca elevadas perdas econômicas em todo mundo, e tem, na tecnologia do DNA recombinante, uma alternativa de desenvolvimento de vacinas mais efetivas. O objetivo do trabalho foi transformar plantas de tabaco para sua utilização como biorreator na produção de um antígeno vacinal contra a PMS. Folhas e entrenós foram cultivados em diferentes concentrações de BAP e AIA. As melhores taxas de regeneração foram encontradas utilizando 1,5 mg.L-1 de BAP e 0,1 mg.L-1 de AIA para segmentos de folhas e entrenós. O teste de seleção utilizando canamicina mostrou-se altamente eficiente, obtendo-se a supressão da regeneração com 30 mg.L-1 e 100 mg.L-1 para segmentos de folhas e entrenós, respectivamente. Colônias recombinantes de A. tumefaciens contendo ltb-r1 foram co-cultivadas com entrenós e segmentos foliares de plantas germinadas in vitro. Após esta etapa, os explantes foram transferidos para meios de seleção, visando selecionar células possivelmente transformadas. O DNA genômico das plantas regeneradas e putativamente transformadas foi extraído e amplificado por PCR, na qual foi possível visualizar uma banda referente ao ltb-r1. A detecção da integração e transcrição do gene foi realizada por Southern blot e RTPCR, respectivamente. Em ambas as técnicas, foi possível verificar a presença de uma banda do tamanho esperado para ltb-r1, demonstrando assim, a integração e expressão do gene. Entretanto, não foi possível detectar com precisão, através dos testes utilizados, a proteína recombinante.

No description available.