Made available in DSpace on 2016-04-28T14:56:19Z (GMT). No. of bitstreams: 1 2009 - Ludmila de Carvalho Sampaio.pdf: 1705521 bytes, checksum: a3e7341ce10bb882edcbb905becd1778 (MD5) Previous issue date: 2009-06-29 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The Restinga is one type of sand coastal ecosystems that are suffering with high anthropic impact and is important the consideration of any action related to the recovery and conservation of the last remaining areas. For that, is necessary to study the grown and the best conditions for seedling production of species that are native from these ecosystems. In this context, the present work had the objectives: 1- study the response of Senna appendiculata and Senna bicapsularis to different sources and amount of nitrogen. 2- study the response of S. appendiculata, Senna bicapsularis and Machaerium lanceolatum to lime and to a changes in the soil pH. The experiments were conduct under greenhouse conditions in pots filled with sand soil with low fertility. For the experiment with different sources and amount of nitrogen, seven treatments were applied: control (without nitrogen), NO3? (10 mg N/ plant), NO3? (20 mg N/plant), NH4+ (10 mg N/plant), NH4+ (20 mg N/plant), NO3NH4 (10mg N/plant) e NO3NH4 (20 mg N/plant). For the experiments with lime and soil pH, four treatments were applied: 0, 2. 4 and 6 t/ha of lime. For each experiment, the following measurement were made: shot and root dry weight, root length, plant height, soil pH, root/shoot ratio, collar diameter and relative growth rate (RGR was estimated only for S .pendula). The results suggest tendency for indicate that the preferential source of nitrogen for S. appendiculata was NH4+. However for S. pendula with the obtained results is not possible to describe the preferential source of nitrogen, although the grown was affected by the nitrogen applied. None of studied plants had the growth limited by acid soil conditions. Senna appendiculata showed preference for growing at low soil pH and the lime was not necessary for the promotion of plant grown. In this conditions if lime is applied can cause a decrease in plant grown. Lime applied showed no effect on the growth of M. lanceolatum. / Devido ao elevado estado de degrada??o dos ecossistemas de restinga ? de extrema import?ncia considerar a??es para a conserva??o e recupera??o dos poucos remanescentes ainda existentes. Para isso faz-se necess?ria a realiza??o de estudos relacionados ao crescimento das esp?cies encontradas nestas ?reas, visando melhores condi??es para a produ??o de mudas. Dentro desta proposta, o presente trabalho teve como objetivos: 1- avaliar o crescimento de Senna appendiculata e Senna bicapsularis, utilizando diferentes fontes e doses de nitrog?nio mineral; e 2- caracterizar a resposta de Senna appendiculata, Senna bicapsularis e Machaerium lanceolatum ? calagem e a mudan?as do pH do solo. Os experimentos foram conduzidos em condi??es de casa de vegeta??o, em vasos contendo como substrato, solo arenoso de baixa fertilidade. Para o experimento de efeito de diferentes doses e fontes de nitrog?nio foram utilizados sete tratamentos: testemunha (sem nitrog?nio), NO3? (10 mg N/planta), NO3? (20 mg N/planta), NH4+ (10 mg N/planta), NH4+ (20 mg N/planta), NO3NH4 (10mg N/planta) e NO3NH4 (20 mg N/planta). No experimento de resposta ? calagem e mudan?a do pH do solo foram utilizados quatro tratamentos: 0, 2, 4 e 6 t de calc?rio/ha. Os par?metros avaliados foram: altura da planta, comprimento da raiz, ac?mulo de mat?ria seca da raiz e da parte a?rea, di?metro do coleto, pH do solo, rela??o raiz/parte a?rea, incremento m?dio em altura (IMA) e taxa de crescimento relativo, este ?ltimo apenas para a esp?cie S. pendula. Os resultados obtidos apresentam uma tend?ncia a indicar que a fonte preferencial de nitrog?nio para a esp?cie S. appendiculata seria o am?nio. O crescimento da esp?cie Senna pendula foi influenciado pela aplica??o de nitrog?nio, no entanto n?o foi poss?vel determinar a fonte de nitrog?nio preferencial. Nenhuma das esp?cies estudadas teve seu crescimento limitado em condi??es de solo ?cido. A esp?cie S. appendiculata demonstrou prefer?ncia por solos ?cidos e a calagem n?o seria necess?ria para a produ??o de mudas desta esp?cie, podendo influenciar negativamente o crescimento da mesma. A calagem n?o influenciou o crescimento de M. lanceolatum.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-05-26T20:35:35Z No. of bitstreams: 1 2012 - Renata Aparecida Costa Silva.pdf: 1687897 bytes, checksum: 4d1af8e117daa1842ae1766319ddf8d6 (MD5) / Made available in DSpace on 2017-05-26T20:35:35Z (GMT). No. of bitstreams: 1 2012 - Renata Aparecida Costa Silva.pdf: 1687897 bytes, checksum: 4d1af8e117daa1842ae1766319ddf8d6 (MD5) Previous issue date: 2012-02-15 / CAPES / Nitrogen is one of the nutrient elements most limiting for plant growth. Thus, increasing plant nitrogen usage efficiency (NUE) is an essential factor for sustainable agriculture, leading to an increased food production with less fertilizer input and less environment impact. The aim of this study was to evaluate the effect of OsDof25 overexpression on N-NO3- and N-NH4+ uptake. In transgenic rice plants, OsDof25 was expressed under control of maize ubiquitin promoter (UBIL:OsDof25:3xHA). Two experiments were conducted: one to evaluate the kinetic parameters Vm?x and KM, and another to analyze the expression level of nitrate (NRT2.1~2.2 and NAR) and ammonium transporters (AMT1.1~1.3), both under high and low NO3- and NH4+ supply. The untransformed plants showed higher growth that transformed lineages. The L1 and L2 showed a lower value of the KM in the resupply treatment of 0.2 mM N-NO3-. In the resupply with 0.2 mM N-NH4 + the L4 showed higher Vmax and L1 lower KM. There were no large variations in uptake kinetics between the transformed and untransformed plants. At the root the NRT2 showed low expression in lineages L1 and L4, when under constant supply of N-NO3-, in contrast, in the treatment under resupply with 0.2 mM N-NO3-was increased expression of OsNTR2.1 ~ 2.2, and NAR in both transformed lineages, but in the root the concentration of NO3- was opposed to the expression of NRT2 and NAR, in both conditions. In the leaves, the line L4 showed high expression of the transporter OsNRT2.1 with the resupply of 0.2 and 2.0 mM N-NO3-. In plants grown under constant supply of N-NH4+, L1 showed lower expression of AMT1 in the root compared to L4 and untransformed plants. When subjected to nitrogen deficiency, there was an increased expression of the OsAMT1.2. However, there was no correlation between N transporter expression levels and NO3- and NH4+ content in the transformed plants, indicating a possible change in enzyme activity and reduction or assimilation of N in these plants. The transformed plants when subjected to resupply with low levels of nitrate and ammonium showed better response parameters Vmax and KM compared to the untransformed. In the plants transformed the resupply with nitrate at low concentration resulted in increasing the gene expression of the transporters (OsNTR2.1 ~ 2.2 and protein OsNAR2.1), and the treatment with constant supply provided greatest nitrate accumulation in these plants. The results of both kinetic parameters and accumulation of fresh matter suggest that plants transformed for the expression of the OsDof25 presented highest tolerance to nutritional stress. / O nitrog?nio ? um dos elementos minerais que mais limita o desenvolvimento das plantas. Assim, aumentar a efici?ncia de uso de nitrog?nio (EUN) ? um fator ? essencial para uma agricultura sustent?vel, levando a um aumento da produ??o de alimentos com menor uso de insumos e menos impactos ao ambiente. Este trabalho teve por objetivo avaliar o efeito da superexpress?o do fator de transcri??o OsDof25 sobre a absor??o de nitrog?nio em duas linhagens transformadas de arroz (L1 e L4) da variedade Nipponbare comparando-as com plantas n?o transformadas (WT). Nas plantas transformadas, o OsDof25 foi expresso sob o controle do promotor da ubiquitina 1 de milho (UBIL:OsDof25:3xHA). For feitos dois experimentos: um para avaliar os par?metros cin?ticos Vm?x e KM, sob condi??es de alto e baixo suprimento de N-NO3- e N-NH4+, e outro para analisar a express?o dos transportadores de NO3- (NRT2.1~2.2 e NAR) e NH4+ (AMT1.1~1.3) tamb?m sob alto e baixo suprimento desses ?ons. As plantas n?o transformadas apresentaram maior crescimento do que as linhagens transformadas. As L1 e a L2 mostraram menor valor de KM no tratamento com ressuprimento de 0,2 mM de N-NO3-. No ressuprimento com 0,2 mM de N-NH4+ a L4 apresentou maior Vm?x e L1 menor KM, mas, n?o houve grandes varia??es nos par?metros cin?ticos de absor??o entre as plantas transformadas e n?o transformadas. Na raiz os NRT2 mostraram baixa express?o nas linhagens L1 e L4 quando submetidas ao suprimento constante de N-NO3-, em contrapartida, no tratamento sob ressuprimento com 0,2 mM de N-NO3- ocorreu aumento de express?o dos OsNTR2.1~2.2 e NAR nas duas linhagens transformadas, por?m na raiz a concentra??o de N-NO3- foi oposta a express?o dos NRT2 e NAR, em ambas as situa??es. Nas folhas, a linhagem L4 apresentou alta express?o do transportador OsNRT2.1 com o ressuprimento de 0,2 e 2,0 mM de N-NO3-. Nas plantas submetidas ao suprimento constante de N-NH4+, a L1 apresentou menor express?o dos AMT1 na raiz quando comparada a L4 e a planta n?o transformada. Quando submetida a defici?ncia de N-NH4+, a express?o do OsAMT1.2 aumentou nas ra?zes de todas as plantas. Entretanto, n?o houve correla??o positiva entre a express?o dos transportadores de N e os teores de NO3- e NH4+ nas linhagens transformadas, indicando uma poss?vel altera??o na atividade das enzimas de redu??o e ou assimila??o de N. As plantas transformadas quando submetidas ao ressuprimento com baixos teores de nitrato e am?nio apresentaram melhor resposta dos par?metros Vm?x e KM em rela??o a n?o transformadas. Nas plantas transformadas o ressuprimento com nitrato em baixa concentra??o resultou em maior express?o dos genes dos transportadores OsNTR2.1~2.2 e da prote?na OsNAR2.1 e o tratamento com suprimento constante proporcionou maior ac?mulo de nitrato nestas plantas. Os resultados tanto dos par?metros cin?ticos quanto do ac?mulo de mat?ria fresca sugerem que as plantas transformadas para express?o do OsDof25 apresentaram maior toler?ncia ao estresse nutricional.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2018-11-09T17:37:20Z No. of bitstreams: 1 2013 - Ana Paula Pessim de Oliveira.pdf: 984469 bytes, checksum: 8daf942927a119ee9cb7fe02201b5631 (MD5) / Made available in DSpace on 2018-11-09T17:37:20Z (GMT). No. of bitstreams: 1 2013 - Ana Paula Pessim de Oliveira.pdf: 984469 bytes, checksum: 8daf942927a119ee9cb7fe02201b5631 (MD5) Previous issue date: 2013-02-27 / CAPES / To evaluate the impacts of trash management on soil quality and sugarcane production in the Brazilian region of coastal tableland three studies were conducted and are presented in distinct chapters. In the first chapter, a literature review, the results of research with sugarcane and changes in biological, chemical and physical soil properties, in response to trash management and their relationship with soil quality, are presented. In general, the data suggest that harvesting without burning affects positively the soil properties and improves soil quality. However, the studies are not sufficient to inform about the effect of mechanical harvesting without burning of trash in the tableland soils. In the second chapter they were evaluated the effect of N fertilization of green sugarcane on the stalk productivity, the trash accumulation, the relationship between the number and weight of stalks, the extraction and accumulation of nutrients in the fresh stalks and trash, and the sugarcane technological quality. This study was conducted in the LASA distillery, in Linhares municipality, Esp?rito Santo State, with ratoon cane, varieties RB86 7515 and RB91 8639, in a Xanthic Udult soil. The experiment was set in a randomized blocks design with the control (without N) and four treatments with N applied as ammonium sulphate (80, 100, 120, and 160 kg N ha-1) in four replicates. The N had a significant effect on stalks productivity. The relationship between the number and weight of stalks was significant for the dosages of 80 kg N ha-1 (both varieties) and 100 kg N ha-1 (only the RB91 8639 variety). There was significant effect of N rates on the cane technological quality only in the second cycle studied. The extractions of N, P and K were significant for the N levels. The third chapter is a simulation study conducted with the APSIM-Sugar cropping systems model, based on data from the log-term experiment (duration of 23 years) in the municipaliy of Linhares-ES. This study had as goal to investigate the impact of trash management after harvesting on sugarcane production and the long-term fate of N contained in trash. The objectives were: to investigate if the model was capable of reproducing the trends of the major N pools (cane yields and soil organic carbon) in the long term experiment; and using the parameterized model to evaluate how the relationship between N fertilizer and yield (and losses of N to the environment) varies with the trash management. The trash management systems conditions simulated were: retention of trash at harvest (100 %, GCTB); half retention of trash at harvest (50%); and burnt cane trash. The N fertilizer management system was simulated by varying N fertilizer application from 0 to 240 kg ha-1 (in 40 kg ha-1 increments) rates on the ratoon crops. The cane yield and soil organic C responded positively to the trash management systems (GCTB and half GCTB), but the magnitude of the response is dependent on N-fertilizer applied on crops. Half removal of trash might reduce the extent of yield increasing potential. When the trash deposition is a recent practice, additional N fertilizer is required by the crop, to avoid yield reduction caused by the N immobilization. The simulations also indicated that the average environmental losses of N were greater from trash retained systems for all rates on N fertilizer applied. / Para avaliar o impacto do manejo da palhada sobre a qualidade do solo e produ??o de cana-de-a??car na regi?o dos Tabuleiros Costeiros, tr?s estudos foram conduzidos e s?o apresentados em cap?tulos. No primeiro cap?tulo, em forma de revis?o de literatura s?o apresentados os resultados de pesquisas com cana-de-a??car e altera??es nas propriedades biol?gicas, qu?micas e f?sicas do solo em resposta ao manejo da palha, e sua rela??o com a qualidade do solo. Do conjunto de dados, se infere que a colheita sem queima com manuten??o da palha altera positivamente as propriedades ed?ficas e melhora a qualidade do solo. Por?m, os estudos s?o insuficientes para suprir informa??es sobre o efeito da colheita mec?nica e sem queima da palhada da cana sobre os solos de tabuleiro. No segundo cap?tulo foram avaliados, os efeitos da aduba??o nitrogenada, em cana crua, sobre a produtividade de colmos, ac?mulo de palhada, rela??o entre n?mero e peso colmo, extra??o e ac?mulo de nutrientes nos colmos frecos e na palhada, e sobre a qualidade tecnol?gica da cana de a??car. O estudo foi realizado na destilaria LASA no munic?pio de Linhares-ES, com cana soca, variedades RB 867515 e RB 918639, em ARGISSOLO AMARELO. Os tratamentos consistiram de cinco doses de N-Sulfato de am?nio (sem N, 80, 100, 120 e 160 kg de N ha-1), arranjados em desenho experimental de blocos casualizados com quatro repeti??es. Houve efeito significativo de doses de N sobre produtividade de colmos. A associa??o entre n?mero e peso de colmos foi significativa para 80 kg N ha-1 (ambas as variedades) e 100 kg ha-1 (para a variedade RB91 8639). Houve efeito significativo das doses de N sobre a qualidade tecnol?gica da cana apenas no segundo ciclo de cultivo estudado. As extra??es dos nutrientes N, P e K mostraram signific?ncia para as doses de N. O terceiro cap?tulo, relata um estudo de simula??o conduzido com o APSM-Sugar cropping systems model, com dados de um experimento de campo de longa dura??o (cerca de 23 anos) em Linhares-ES. Este estudo se prop?s a investigar o impacto do manejo da palhada ap?s a colheita sobre a produtividade da cana e o destino em longo prazo do N contido na palha. Os objetivos foram: investigar se o modelo foi capaz de reproduzir as tend?ncias de maiores ?pools? de N (rendimento de cana e C org?nico no solo) no experimento de longa dura??o; e usar o modelo parametrizado para investigar como a rela??o entre o N-fertilizante e rendimento (e perdas de N para o ambiente) varia com o manejo da palhada. As condi??es de manejo da palha simuladas foram: 100% e 50% da palhada retida ap?s a colheita; e palhada queimada. O manejo da aduba??o foi simulado para variar com as doses de N de 0 a 240 kg ha-1 (com incrementos de 40 kg ha-1) em cana soca. O rendimento de cana e o C org?nico do solo foram bem representados pelo modelo. O rendimento de cana respondeu positivamente aos sistemas de manejo da palhada (100% e 50% de reten??o), mas a magnitude da resposta depende da aduba??o nitrogenada. A remo??o de 50% da palha pode reduzir o efeito de rendimento potencial. Quando a deposi??o da palhada ? pr?tica recente, N adicional ? requerido pela cultura para evitar redu??o do rendimento causada pela imobiliza??o do N. As simula??es tamb?m indicam que as perdas ambientais de N foram maiores nos sistemas com cana crua para todas as doses de N.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-07-26T18:47:21Z No. of bitstreams: 1 2014 - Marcela Jacques de Lemos Neves.pdf: 434472 bytes, checksum: c9817e97c5d136299acb936ad4b87b6f (MD5) / Made available in DSpace on 2017-07-26T18:47:21Z (GMT). No. of bitstreams: 1 2014 - Marcela Jacques de Lemos Neves.pdf: 434472 bytes, checksum: c9817e97c5d136299acb936ad4b87b6f (MD5) Previous issue date: 2014-05-30 / FAPERJ / The main goal of this study was evaluate the effect of downregulation of the ammonium transporter OsAMT1.3 in the ammonium uptake through the high affinity system, as well as the effects on nitrogen metabolism. To perform the OsAMT1.3 gene silencing, it was used the artificial micro RNA (amiRNA) technology. In this system, the OsAMT1.3 coding region sequence (cds) is inserted in W marcelaMD3 website and putatives amiRNAs to silencing OsAMT1.3 were made. The amiRNA was inserted by PCR using the pNW55 vector as template. The amiRNA was inserted in the IRS154 vector using the T4 DNA ligase. The IRS154 plus amiRNA was cloned in the E. coli by electroporation. After rice transformation of Nipponbare variety through Agrobacterium and Hygromycin selection, 14 lineages were obtained. Six lineages showed high seed production and only one lineage with abnormal growth. After seed production in a greenhouse, the lineages L1, L2 and L6 were selected to further experiments evaluating the effects of OsAMT1.3 downregulation. First, the lineages selected were evaluated about the levels of OsAMT1.3 downregulation. The rice lineages transformed with amiRNA showed lower level of OsAMT1.3 expression compared to control plants, however, different levels of OsAMT1.3 downregulation was observed. The lineage L1 showed lower levels of OsAMT1.3 downregulation, L2 and L6 showed higher levels of OsAMT1.3 downregulation. The lineages L1, L2 and L6 as well as IRS control plant (transformed with empty vector) were grown in growth chamber at 30 days after germination, and the treatments used were: N starvation for three days, resupply with 0.15 mM of NH4+-N (low level) and 2.0 mM of NH4+-N (high level). The lineages L1, L2 and L6 showed lower NH4+ uptake with 0.15 mM of NH4+-N compared to control plants (IRS), on the other hand, the plants grown with 2.0 mM of of NH4+-N did not show NH4+ uptake differences, except L1. The expression of OsAMT1.1, OsAMT1.2 and OsAMT1.3 ammonium transporters were upregulated with 0.15 mM of NH4+-N in the control plants (IRS); in the lineages L1, L2 and L6 showed downregulation of the OsAMT1.1, OsAMT1.2 and OsAMT1.3 genes. The lower NH4+ uptake with 0.15 mM of NH4+-N resulted in lower levels of NH4+-N and Amino-N in the roots in the lineages, while the NH4+-N and Amino-N in the plants grown with 2.0 mM of NH4+-N was minimally changed. The results indicate that OsAMT1.3 downregulation leads to OsAMT1.1 and OsAMT1.2 downregulation as well, decreasing the NH4+ uptake. Despite the OsAMT1.3 lower expression compared to OsAMT1.1 and OsAMT1.2, the OsAMT1.3 transporter may be involved in the nitrogen uptake efficiency in low levels of NH4+. / O objetivo deste trabalho foi estudar o efeito do silenciamento do gene do transportador OsAMT1.3 em arroz sobre a habilidade das plantas em absorver o N-NH4+ atrav?s do sistema de alta afinidade, bem como os reflexos sobre o metabolismo de nitrog?nio. Para o silenciamento do gene OsAMT1.3 foi usada a tecnologia do miRNA artificial (amiRNA). Para tanto, a sequ?ncia codante do gene (cds) OsAMT1.3 ? inserida no sistema WMD3 que indica sequ?ncias de amiRNA?s para silenciar o pr?prio OsAMT1.3. A inser??o do amiRNA foi feita por PCR usando o vetor pNW55 como molde. O miRNA foi inserido no vetor IRS154 por corte e liga??o usando a enzima T4 DNA ligase. O produto da liga??o foi introduzido em E. coli por eletropora??o. Ap?s a transforma??o de arroz da variedade Nipponbare mediada por Agrobacterium. A sele??o das plantas transformadas foi feita com o antibi?tico Higromicina. No total foram obtidas 14 linhagens transformadas. Para confirmar que as plantas mutantes possuiam a constru??o, foi realizado o teste com a folha bandeira em solu??o de higromicina. Seis linhagens apresentaram boa produ??o de sementes e houve uma linhagem com crescimento anormal. Ap?s a multiplica??o das linhagens em casa de vegeta??o, foram selecionadas as linhagens L1, L2 e L6 para os experimentos de an?lise dos efeitos do silenciamento do gene OsAMT1.3. Primeiramente, as linhagens selecionadas foram avaliadas quanto ao n?vel de silenciamento do gene OsAMT1.3. As linhagens de arroz transformadas apresentaram maior n?vel de silenciamento do gene OsAMT1.3 quando comparadas ?s plantas controle, no entanto, diferentes n?veis de silenciamento foram observados. A linhagem L1 apresentou menor n?vel de silenciamento do gene OsAMT1.3, enquanto L2 e L6 apresentaram maior silenciamento. As linhagens L1, L2 e L6 e a planta controle IRS (transformada com o vetor vazio) foram cultivadas em c?mara de crescimento at? os 30 dias ap?s a germina??o, com a aplica??o dos seguintes tratamentos: sem N por tr?s dias, ressuprimento com 0,15 mM de N-NH4+ ap?s tr?s dias de priva??o de N (baixa dose) e 2,0 mM de N-NH4+ constante (alta dose). As linhagens L1, L2 e L6 apresentaram menor absor??o de NH4+ com 0,15 mM de N-NH4+ quando comparadas com as plantas controle (IRS), enquanto as plantas cultivadas com 2,0 mM de N-NH4+ n?o apresentaram diferen?as na absor??o de NH4+. A express?o dos genes dos transportadores de NH4+ OsAMT1.1, OsAMT1.2 e OsAMT1.3 foi induzido pelo tratamento com 0,15 mM de N-NH4+ nas plantas controle (IRS), enquanto as linhagens transformadas apresentaram repress?o dos genes OsAMT1.1, OsAMT1.2 e OsAMT1.3. A menor absor??o de NH4+ com 0,15 mM de N-NH4+ causou menor n?vel de N-NH4+ e N-amino nas ra?zes das linhagens transformadas, enquanto nas plantas com 2,0 mM de N-NH4+ houve pouca altera??o nos conte?dos de N-NH4+ e N-amino. Os resultados indicam que o silenciamento do gene OsAMT1.3 provoca regula??o negativa dos transportadores OsAMT1.1 e OsAMT1.2, alterando a absor??o de NH4+. Apesar do gene OsAMT1.3 ser menos expresso que os genes OsAMT1.1 e OsAMT1.2, o transportador OsAMT1.3 pode estar envolvido na efici?ncia de absor??o em baixas doses de NH4+.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2017-05-26T20:53:49Z No. of bitstreams: 1 2010 - Joice de Jesus Lemos.pdf: 529861 bytes, checksum: 869aef193d0f1b7b1b385f7ee3a2de73 (MD5) / Made available in DSpace on 2017-05-26T20:53:49Z (GMT). No. of bitstreams: 1 2010 - Joice de Jesus Lemos.pdf: 529861 bytes, checksum: 869aef193d0f1b7b1b385f7ee3a2de73 (MD5) Previous issue date: 2010-08-31 / CAPES / With the aims to study the efflux of H+ or OH- due nitrogen sources on the process of infection of Fusarium oxysporum f. sp. phaseoli in the rhizosphere of two beans cultivars Diamante Negro (susceptible) and Ouro Negro (more resistant), a series of experiments were done in greenhouse and growth chamber. Were applied to three different sources of nitrogen (N-N2 N-NO3- and N-NH4+) in three nitrogen concentrations (0, 30 and 120 kg ha-1), in plants grown in clay and sand soil. In addition, an experiment was conducted with different concentrations of inoculum of Fusarium (0, 10 ? and 106 conidia mL-1) in greenhouse in order to know the concentration that would affect the infection and when applied the fungi. Overall, the results suggested that nitrate decreased the infection process of Fusarium and ammonium increase. The association between nitrate with the cultivar more tolerant Ouro Negro, decreased the perceptual infection of fungi. The source of nitrogen influence of pH of rhizosphere occurred interaction with the type of soil. In all the experiments, found lower numbers of nodules. The concentration of inoculum or the times of inoculation not produce effect in the perceptual of infection. / Foram realizados estudos com duas cultivares de feijoeiro comum (Phaseolus vulgaris L.), a Diamante Negro, considerada suscet?vel ao Fusarium oxysporum f. sp. phaseoli e a Ouro Negro, mais resistente a esse fungo causador da murcha-de-fus?rio. Os experimentos foram instalados em c?mara crescimento ou em casa de vegeta??o. O objetivo deste trabalho foi o de analisar a influ?ncia da libera??o OH- ou H+ devido ao uso de fontes nitrogenadas na infec??o do fungo. Foram utilizadas tr?s fontes de nitrog?nio (N-N2, N-NO3 ? e N-NH4+) e diferentes doses de nitrog?nio (0, 30 e 120 kg ha-1) com plantas inoculadas com o referido fungo, crescidas no substrato areia ou em solos com diferentes teores de argila. Tamb?m foi realizado um experimento utilizando diferentes concentra??es de in?culo do Fusarium (0, 10? e 106 con?dios mL-1) com o objetivo de analisar qual concentra??o afetaria mais a infec??o na presen?a de fontes nitrogenadas. Foram analisados, o percentual de infec??o do Fusarium, o pH da rizosfera e n?o rizosf?rico, massas da parte a?rea e ra?zes secas, e o n?mero de n?dulos em diferentes ?pocas de amostragem. De modo geral foi observado que a fonte nitrato diminuiu o processo de infec??o do Fusarium e a fonte am?nio aumentou. Foi confirmado que a cultivar Ouro Negro ? mais tolerante ao fungo e que quando associada ? fonte de nitrog?nio nitrato aumentou ainda mais a resist?ncia. O pH da rizosfera e n?o rizosf?rico foram influenciados pela fonte de nitrog?nio: nitrato aumenta, e am?nio diminui. Os dados do trabalho sugeriram haver intera??o entre a fonte de nitrog?nio x dose x cultivar x solo. O n?mero de n?dulos encontrado nas condi??es experimentais foi baixo, especialmente nas amostragens na fase inicial do ciclo.

Made available in DSpace on 2016-04-26T19:39:32Z (GMT). No. of bitstreams: 1 2009 - Leandro Azevedo Santos.pdf: 3069700 bytes, checksum: dbd1726a5b683e7f290e1829143eacac (MD5) Previous issue date: 2009-06-03 / Funda??o Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro / To improve nitrogen usage efficiency in plants the rice transcriptional factor OsDof25 was identified and cloned, whose probably orthologe is the maize ZmDof1, already identified and partially characterized. The ZmDof1 was also cloned for comparative analysis with OsDof25, in order to confirm this last one as ZmDof1 orthologe in rice. The constructions for Arabidopsis superexpression of these transcriptional factors were made using the cloning system of gateway technology (Invitrogen), to obtain the expression vectors 35S:ZmDof1:HA and 35S:OsDof25:HA. Lineages with different expression levels of these genes were obtained, but with only one inserted copy. These transgenic lineages when grown in a half strength of MS medium (10mM of NH4 + and 20mM of NO3 -) showed phenotypes with chloroses and growth difficulty; although when they were cultured in soil they showed great vegetative development and delay in the inflorescence emission. When analyzed the gene expression changes induced by the superexpression of these transcriptional factors, it was observed that both genes produced an increase in the expression levels of high and low affinity ammonium transporters (AMT1.1 and AMT2.1, respectively), indicating that these phenotypes may be due to the toxic effect of an excess of ammonium uptake. We also verified an increase of expression for pyruvate kinase (PK1 and PK2), and phosphoenolpyruvate carboxylase (PEPC1 and PEPC2). Pyruvate kinase converts phophoenolpyruvate (PEP) to pyruvate, and phosphoenolpyruvate carboxylase converts PEP to oxalacetate, which is substrate for malate dehydrogenase to form malate. Both pyruvate and malate may feed the Krebs cycle. In addition, there was an increase in the expression of isocitrate dehydrogenase, which is present in the citosol and mitochondria, needed for converting isocitrate to 2- oxoglutarate. Thus, it was hypothesized that the increase of expression levels of these carbon metabolism enzymes was necessary to increase the production of 2-oxoglutarate and, consequently, to reduce the toxic effect of ammonium uptaked. Besides, it was observed an increase of expression levels and activity of glutamate dehydrogenase (GDH). This enzime may work as much in the direction of glutamate amination as in deamination, when the plants were submitted to ammonium excess or carbon limitation conditions, respectively. / Com o objetivo de aumentar a efici?ncia de uso de nitrog?nio (EUN) em plantas, foi identificado e clonado o fator de transcri??o OsDof25 de arroz, cujo prov?vel ort?logo ? o ZmDof1 de milho, j? identificado e parcialmente caracterizado. Tamb?m foi clonado o ZmDof1 para an?lises comparativas com o OsDof25, a fim de comprovar que este ?ltimo ? realmente ort?logo do ZmDof1. As constru??es para superexpress?o destes fatores de transcri??o em Arabidopis foram feitas utilizando o sistema gateway de clonagem para obten??o dos vetores de express?o 35S:ZmDof1:HA e 35S:OsDof25:HA. Foram obtidas linhagens com diferentes n?veis de express?o destes genes, mas com apenas uma inser??o. As linhagens transg?nicas obtidas quando crescidas em meio MS ? for?a i?nica (10mM de NH4 + e 20mM de NO3 -) apresentaram fen?tipos como clorose e dificuldade de desenvolvimento, ao passo que quando cultivadas em solo mostraram desenvolvimento vegetativo mais intenso e atraso para emiss?o da infloresc?ncia. Quando analisadas as modifica??es de express?o g?nica causadas pela superexpress?o destes fatores de transcri??o, observou-se que ambos os fatores de transcri??o provocaram aumento de express?o dos transportadores de am?nio de alta e baixa afinidades (AMT1.1 e AMT2.1 respectivamente), indicando que o fen?tipo observado pode ser devido ao efeito t?xico do excesso de am?nio absorvido. Verificou-se tamb?m aumento de express?o das enzimas piruvato quinase (PK1 e PK2) e fosfoenolpiruvato carboxilase (PEPC1 e PEPC2). A piruvato quinase converte o fosfoenolpurato (PEP) a piruvato, enquanto a fosfoenolpiruvato carboxilase converte o PEP a oxalacetato (OAA) que pode sofrer a??o da malato desidrogenase originando o malato. Ambos os metab?litos, piruvato e malato, alimentam o ciclo de Krebs. Houve tamb?m aumento de express?o da isocitrato desidrogenase, enzima presente na mitoc?ndria (ciclo de Krebs) e no citosol que converte isocitrato a 2-oxoglutarato (2-OG). Assim, ? prov?vel que o aumento da express?o destas enzimas do metabolismo de carbono foi necess?rio para aumentar a produ??o de 2-OG e, por conseguinte, diminuir o efeito t?xico do excesso de am?nio absorvido. Al?m disso, observou-se aumento de express?o e atividade da glutamato desidrogenase (GDH). Essa enzima pode atuar tanto na dire??o da amina??o, quanto na dire??o da desamina??o, em condi??es de excesso de am?nio e/ou sob condi??es de limita??o de carbono nas plantas, respectivamente.