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The disease vectors Aedes aegypti and Aedes albopictus are serious and growing threats to global health. As vectors of the arboviruses dengue fever and chikungunya, these mosquitoes are responsible for hundreds of millions of cases and thousands of deaths each year. Absent specific treatments or vaccines, effective control of mosquito populations remains the only option for tackling a growing public health challenge. More effective control tools are urgently needed. Recently, a novel approach to pest population control has been developed based on the release of insects carrying a repressible, dominant lethal allele. This approach has achieved dramatic reductions in Ae. aegypti populations in regulated open field experiments. Despite this success, there remains scope to improve upon the current technology. It is proposed that an 'ideal' strain would combine the following features: (i) repressible lethality in late juvenile phases; (ii) a mechanism for removing females at an early developmental stage in the release generation; and (iii) orthogonal expression control mechanisms allowing both these systems to be combined in a single strain. This thesis describes research undertaken in pursuit of a 'next generation' strain. Two novel promoters from putative Osiris genes have been identified which confer a 10²-10³ – fold up-regulation in transgene expression specific to late pupal stages. One of these 'Osiris' promoters has been used to develop transgenic Aedes aegypti strains. 5 lines showed pupal-specific lethality of 98-100% penetrance, which was repressed in the presence of tetracycline. An Ae. albopictus orthologue of the sex-determining gene doublesex (dsx) has been isolated and characterised and a female-specific expression system developed. Transgenic lines show female-specific expression of a transgene; however, there remains some 'leaky' expression in male insects. Finally, a potential expression control tool based on an auxin-inducible expression system has been investigated. 11 different transgenic lines were developed based on three different construct designs. None showed auxin-inducible expression of a transgene.

Cassava brown streak disease (CBSD) threatens food and economic security for smallholder farmers throughout East and Central Africa, and poses a threat to cassava production in West Africa. CBSD is caused by two whitefly-transmitted virus species: Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV) (Genus: Ipomovirus, Family Potyviridae). Although varying levels of tolerance have been achieved through conventional breeding, to date, effective resistance to CBSD within East African cassava germplasm has not been identified. RNAi technology was utilized to integrate CBSD resistance into the Ugandan farmer-preferred cassava cultivar TME 204. Transgenic plant lines were generated expressing an inverted repeat construct (p5001) derived from coat-protein (CP) sequences of CBSV and UCBSV fused in tandem. Northern blots using probes specific for each CP sequence were performed to characterize 169 independent transgenic lines for accumulation of CP-derived siRNAs. Transgenic plant lines accumulating low, medium and high levels of siRNAs were bud graft challenged with the virulent CBSV Naliendele isolate alone or in combination with UCBSV. Resistance to CBSD in the greenhouse directly correlated to levels of CP-derived siRNAs as determined by visual assessment of leaf and storage root symptoms, and RT-PCR diagnosis for presence of the pathogens. Low expressing lines were found to be susceptible to CBSV and UCBSV, while medium to high accumulating plant lines were resistant to both virus species. Absence of detectable virus in the best performing p5001 transgenic lines was further confirmed by back-inoculation via sap or graft challenge to CBSD susceptible Nicotiana benthamiana and cassava cultivar 60444, respectively. Data presented shows robust resistance of transgenic p5001 TME 204 lines to both CBSV and UCBSV under greenhouse conditions. Levels of resistance correlated directly with levels of transgene derived siRNA expression such that the latter can be used as predictor of resistance to CBSD.

Cardiac fibrosis contributes to heart failure by dramatically impairing cardiac function, increasing patient morbidity and mortality. The primary fibrillar collagen expressed in the heart is type I, and increased collagen synthesis is the hallmark of cardiac fibrosis. Our laboratory has shown that the transcription factor scleraxis is sufficient to regulate the gene encoding collagen Iα2. The present thesis identifies and focuses on three key functions of scleraxis in the heart. First, we show that scleraxis is required for production of the cardiac extracellular matrix. Using in vitro and in vivo models, we observed a significant upregulation/reduction of matrix genes in response to induction/loss of scleraxis gene function respectively. In fact, scleraxis overexpression was sufficient to rescue matrix synthesis in scleraxis-null cells. In a murine model of cardiac pressure overload, scleraxis gene deletion blunted the induction of fibrotic collagen gene expression. Second, we provide evidence that scleraxis governs fibroblast-myofibroblast phenotype transition and fibroblast number. Scleraxis gene induction promoted cardiac myofibroblast phenoconversion while knockdown reduced myofibroblast marker gene expression. Scleraxis exerts direct transcriptional control on the a-smooth muscle actin gene-an established marker of myofibroblasts. Scleraxis null mice exhibited a dramatic reduction in cardiac fibroblast numbers- this is attributed to impairment of the epithelial-to-mesenchymal transition program which was marked by a corresponding loss of mesenchymal markers and increased epithelial markers. Loss-of-function experiments using primary cardiac proto-myofibroblasts recapitulated this paradigm, whereas scleraxis gene induction showed a reciprocal effect on mesenchymal markers. Third, data from this study supports the required role of scleraxis in the TGFb/Smad signaling pathway. Scleraxis is strongly upregulated by the potent pro-fibrotic cytokine TGFb, and works synergistically with the canonical Smad signaling pathway to increase Col1a2 expression by cardiac fibroblasts and myofibroblasts. Smad3 induced expression of the fibrillar collagens – an effect that was significantly attenuated following scleraxis knockdown. Smad3 binding to the Col1a2 gene promoter was significantly reduced in scleraxis null hearts. This study involved a comprehensive series of in vitro and in vivo experiments, and is the first to identify scleraxis as a key regulator of multiple fibroblast functions and a potential future target for therapeutic intervention in cardiac fibrosis. / May 2016

La maladie de Charcot-Marie-Tooth liée à l'X (CMTX), deuxième cause, en fréquence, de neuropathies héréditaire, est due à des mutations dans le gène Gjb1 codant pour la connexine 32. Afin de les utiliser comme modèle pré clinique, nous avons créé 5 lignées de souris transgéniques, ayant intégré un BAC humain portant une mutation observée dans plusieurs familles indépendantes. L'exploration de ces modèles a montré que la connexine 32 (Cx32) est impliquée dans le contrôle de la stabilité mitotique. Nous avons ensuite montré que cette instabilité implique l'activité des CaMKII et, peut être, de la kinase Pim1. Cette instabilité est corrigée par des inhibiteurs des CaMKII (KN62 et KN93). Nous avons retrouvé le même phénomène dans des cellules de malades CMTX. Nous avons également pu montrer que les animaux transgéniques montrent des anomalies du comportement locomoteur, corrigées par un traitement par des inhibiteurs de CaMKII. Finalement, nous proposons des pistes pour améliorer ces molécules, en synthétisant des analogues de KN93 / X-linked Charcot -Marie -Tooth (CMTX) disease, the second cause, in frequency, of hereditary neuropathies, is caused by mutations in the gene GJB1 encoding connexin 32. As a preclinical model, we created five lines of transgenic mice, which have integrated a human BAC contain mutation observed in several independent families. The exploration of models showed that the connexin 32 (Cx32) is involved in the control of mitotic stability. We then showed that this instability involves the activity of CaMKII and, perhaps, kinase Pim1. This instability is corrected by inhibitors of CaMKII (KN62 and KN93). We found the same phenomenon occuring in cells of CMTX patients. We also showed that transgenic animals show abnormal locomotor behavior corrected by treatment with inhibitors of CaMKII. Finally, we propose strategies to improve efficiency of these molecules by synthesizing analogues of KN93

Orientador: Ivan de Godoy Maia / Banca: Paulo Eduardo Martins Ribolla / Banca: Marcelo Menossi Teixeira / Resumo: As proteínas desacopladoras pertencem à família de carreadores aniônicos mitocondriais. De maneira geral, as proteínas desacopladoras dissipam o gradiente eletroquímico de prótons gerados na respiração na forma de calor, sendo dependentes de ácidos graxos e sensíveis aos nucleotídeos purínicos. O presente estudo visou investigar o comportamento de plantas transgênicas de tabaco que expressam de forma constitutiva o gene AtUCP1, frente aos estresses osmótico e salino, bem como analisar a atividade das regiões promotoras dos genes AtUCP1 e AtUCP2 de Arabidopsis thaliana, em resposta aos estresses osmótico e de baixa temperatura, e ao ácido abscísico. Numa primeira abordagem foram utilizadas sementes selvagens e de duas linhagens transgênicas, germinadas em meio MS adicionados ou não de NaCl e Manitol. O teste de germinação revelou que as linhagens transgênicas apresentam uma maior tolerância aos referidos estresses. Quando o crescimento radicular foi analisado, uma maior inibição foi constatada no controle não transgênico em relação às duas linhagens transgênicas testadas. Adicionalmente, quando submetidas aos estresses, uma maior acumulação de ânion superóxido foi verificada nas folhas de plântulas não transgênicas em relação às plântulas das linhagens transgênicas. Quanto à quantificação de GUS nas plantas transformadas com os promotores dos genes AtUCP1 e AtUCP2, nenhuma alteração significativa foi observada em nenhum dos tratamentos testados / Abstract: The uncoupling proteins belong to the mitochondrial anion carrier family. In general, the uncoupling proteins dissipate the proton electrochemical gradient generated in respiration as heat, being dependent on fatty acids and sensitive to purine nucleotides. In the present study, we investigated the behavior of transgenic tobacco plants that overexpress the AtUCP1 gene when subjected to osmotic and saline stress, as well as the activity of the promoters of the AtUCP1 and AtUCP2 genes of Arabidopsis thaliana, in response to osmotic and cold stress, and abscisic acid. In the first approach, seeds from wild type and two transgenic lines were germinated in MS medium containing (or not) NaCl and mannitol. The germination test showed that the transgenic lines have a higher stress tolerance. When root growth was analyzed, a greater inhibition was observed in non-transgenic control seedlings as compared to seedlings of the two transgenic lines tested. Additionally, when subjected to stress, a greater superoxide anion accumulation was detected in leaves of non-transgenic seedlings as compared to seedlings of transgenic lines. Quantification of GUS activity in the plants transformed with the tested promoters, revealed no treatmentspecific differences / Mestre

No presente trabalho foi estudado o efeito do consumo de soja geneticamente modificada e de sua parental em parâmetros nutricionais e hematológicos em ratos em crescimento. Farinha de soja, desengordurada e autoclavada, foi incorporada em rações experimentais: 12% de proteína para o 1° ensaio e 10% para 2° ensaio. As rações com soja foram suplementadas com aminoácidos essenciais (Ieucina, lisina, metionina e vali na) no 2° ensaio, devido a menor concentração desses aminoácidos nas rações do 1° ensaio quando comparadas ao grupo controle, perfil confirmado pelo aminograma. Ratos Wistar (n=64) foram alimentados, ad libitum, com rações controle (AIN-93G) e rações com a farinha de soja por 28 (1° ensaio) e 32 dias (2° ensaio). Para avaliação do aproveitamento biológico da proteína da dieta e da qualidade protéica, foram realizadas análises de nitrogênio na carcaça dos animais, nas fezes e na urina que foram coletadas ao longo de 2 períodos de balanço de 6 dias cada ensaio. Os parâmetros nutricionais avaliados foram : Coeficiente de eficácia alimentar (CEA), Coeficiente de eficácia protéica (PER), Coeficiente de eficiência líquida da proteína (NPR), Valor biológico da proteína (BV), Coeficiente de utilização líquida da proteína (NPU), digestibilidade protéica e o \"Escore químico corrigido pela digestibilidade real da proteína\". Para avaliação do estado nutricional foram avaliadas as concentrações de albumina, proteínas totais e IGF-1 no plasma. Como parâmetros hematológicos, foram avaliados o hemograma completo, mielograma e esplenograma. Os resultados obtidos demonstraram que animais alimentados com soja GM e sua parental apresentaram desenvolvimento semelhante ao grupo controle (caseína). Os valores de CEP e NPR encontrados nos 2 ensaios mostram que a proteína de soja é nutricionalmente adequada. A oferta protéica de 10% com suplementação com aminoácidos essenciais favoreceu no desenvolvimento dos animais, visto que foi observado aumento de peso em média de 50g, PER e NPR melhores quando comparados com a oferta protéica de 12% sem suplementação. Nos dois ensaios, os valores de digestibilidade dos grupos alimentados com soja foram inferiores aos do grupo controle, resultado evidenciado pela maior dificuldade das enzimas digestivas em digerirem a proteína vegetal, maior perda de aminoácidos endógenos e quantidade de fibra insolúvel presente na soja. O valor de POCAA de 85% demonstra bom aproveitamento dos grupos com soja. Não houve diferença estatística nos parâmetros plasmáticos estudados, bem como nos resultados hematológicos sugerindo que o consumo de soja, convencional ou GM, não altera o estado nutricional dos animais. / In the present work, the effects of genetically modified (GM) soybean and its parental on nutritional and hematological parameters in growing rats were studied. Autoclaved, fat-free soybean flour was added to experimental diets: 12% protein in the first assay and 10% in the second assay. The soybean diets were supplemented with essential amino acids (Ieucine, Iysine, methionine and valine) in the second assay, since such amino acids presented a lower concentration in the diets of the first experiment, as confirmed by the aminogram. Wistar rats (n= 64) were fed a control diet (AIN-93G) and diets supplemented with soybean flour (parental and GM) ad libitum over 28 days (1st experiment) and 32 days (2nd experiment). For evaluating the biological utilization of dietary protein and protein quality, nitrogen analyses were carried out on the animal carcasses, feces and urine. Feces and urine were collected along two 6-day periods within each assay. The following nutritional parameters were evaluated: feed efficiency ratio (FER), protein efficiency ratio (PER), net protein ratio (NPR), biological value (BV), net protein utilization (NPU), protein digestibility and protein digestibility corrected by amino acid (POCM) score. For evaluating the nutritional status, plasma albumin, total proteins and IGF-1 were analysed. Complete hemogram, myelogram and splenogram were used as hematological parameters. The results indicate that animals fed the GM soybean and its parental showed a similar growth rate to the control group (casein). PER and NPR values found in both assays show that soybean protein is nutritionally adequate. A diet containing 10% protein supplemented with essential amino acids favored the growth of the animals in comparison to a diet containing 12% protein without amino acid supplementation. In both assays, the protein digestibility in soybean-fed groups was lower than in the control group, a result evidenced by a greater difficulty of enzymes in digesting vegetal proteins, a greater loss of endogenous amino acids and the presence of insoluble dietary fiber in the soybean. A POCM value of 85% showed a good protein utilization in the soybean-fed groups. No statistically significant differences were observed in the plasmatic and hematological parameters among the groups, suggesting that consumption of soybean (either GM or parental) does not alter the nutritional status of the animals.

Muitos fatores, como variações sazonais, tipos de tecido vegetal, cultivares e espécies de hospedeiro, tipo de solo, interação com microrganismos benéficos ou patógenos entre outros, afetam a estrutura e a composição da comunidade bacteriana das plantas. A introdução de plantas geneticamente modificadas (PGM) foi somada ao conjunto desses fatores, podendo acarretar efeitos diretos e indiretos sobre a comunidade bacteriana. Cana-de-açúcar é uma cultura de grande importância no Brasil; a área de cultivo está em expansão devido aos incentivos para a produção de álcool. Uma bactéria potencialmente importante para a cultura de cana-de-açúcar é a Methylobacterium, um importante endófito de diversas culturas de interesse econômico e que, em cana-de-açúcar, pode melhorar a germinação de sementes, promover um aumento do peso da planta e da área foliar, além do número de internódios. Dessa forma, o trabalho teve como objetivos: i) avaliar os efeitos da canade- açúcar transgênica resistente a insetos e a herbicida sobre a comunidade bacteriana associada; ii) avaliar se o suposto efeito causado pela cana-de-açúcar transgênica resistente a herbicida deriva diretamente do transgene ou dos tratos culturais diferenciados aos quais a planta transgênica é submetida; iii) avaliar a interação entre Methylobacterium spp. e cana-de-açúcar. Analisando a densidade bacteriana das plantas de cana-de-açúcar convencionais e transgênicas não foi possível constatar diferenças relacionadas à introdução dessas plantas. Analisando somente quanto ao manejo de ambos os experimentos, foi possível observar que diferenças em trato cultural ou manejo de plantas, decorrentes ou não da transgenia, podem influenciar a comunidade bacteriana. Por meio de ARDRA, foi possível observar distribuição diferenciada dos ribotipos com a introdução das PGMs. No experimento realizado somente com plantas transgênicas para resistência a Imazapyr, foi possível notar que, em 17 meses, a presença da planta transgênica e a aplicação do herbicida Imazapyr podem ter resultado na redução da densidade bacteriana associada à rizosfera de cana-de-açúcar. Por meio de DGGE, foi visto que o estado fisiológico da planta foi a maior fonte de variação. Novamente por meio de ARDRA, foram observados ribotipos cuja presença foi afetada pelo cultivo da planta transgênica. Esses ribotipos diferentemente distribuídos poderiam resultar em alterações na atividade bacteriana dessas plantas, uma vez que esses ribotipos podem representar grupos funcionais importantes. A colonização de Methylobacterium spp. em cana-de-açúcar foi analisada por meio de microscopia eletrônica de varredura, reisolamento e microscopia óptica de fluorescência. Foi visto que as linhagens utilizadas colonizam cana-de-açúcar, sendo que os pontos de maior colonização são os flanges cuticulares e as regiões pilosas da raiz. Outros estudos são necessários para o melhor aproveitamento dessa bactéria na cultura de cana-de-açúcar. / Many factors, such as seasonal variations, kinds of vegetal tissue, cultivares and host species, kind of soil, interaction with beneficial or pathogen microorganisms, among others, affect the structure and composition of plants bacterial community. The introduction of genetically modified plants (GMP) was added to the set of these factors, making it possible to cause direct and indirect effects on the bacterial community. Sugarcane is a very important crop in Brazil; the cultivation area is expanding due to incentives to alcohol production. A potentially important bacterium for sugar cane cultivation is the Methylobacterium, which is an important endophyte for several cultures of economic interest and which can improve seed germination in sugar cane, promote an increase of plant weight and foliar area, and also the internodes. Thus, the work had as objectives: i) to assess the effects of transgenic sugarcane resistant to insects and herbicide on the associated bacterial community; ii) to assess if the presumed effect caused by transgenic sugarcane resistant to herbicides derives directly from the transgene or differentiated cultural handlings to which the genetically modified plant is undertaken; iii) to assess the interaction between Methylobacterium ssp. and sugarcane. Analyzing the bacterial density of conventional and transgenic sugarcane plants it was not possible to see differences related to the introduction of these plants. Analyzing only in relation to the handling of both experiments, it was possible to see that differences in cultural wielding or handling of plants derived or not from transgenia can influence the bacterial community. By means of ARDRA, it was possible to see a differentiated distribution of ribotypes with the introduction of GMPs. In the experiment made only with transgenic plants for Imazapyr resistance, it was possible to see that in 17 months, the presence of the transgenic plant and the application of Imazapyr herbicide can bring result regarding the bacterial density reduction associated to sugarcane rizosphere. By means of DGGE, it was seen that the physiological status of the plant was the greatest variation source. Again, by means of ARDRA, ribotypes whose presence was affected by transgenic plant cultivation were observed. If distributed differently, these ribotypes can represent important functional groups. The Methylobacterium ssp. Colonization in sugarcane was analyzed and by means of scanning electronic microscopy, re-isolation, and fluorescence optical microscopy. It was observed that the utilized lineages colonize sugarcane being cuticle flange and perilous regions of root the highest colonization points. Other studies are needed to a better good use of this bacterium in sugarcane culture.

Com o aumento progressivo do número de casos de infecção por diferentes arbovírus, por exemplo, dengue, zica e chikungunya, faz-se necessário o desenvolvimento de novas técnicas para o combate a esses arbovírus. A manipulação genética possibilitou a obtenção de mosquitos geneticamente modificados que sejam capazes de suprimir a população selvagem ou impedir a transmissão de agentes etiológicos gerando doenças. O estudo teve como objetivo o estabelecimento de linhagens para supressão populacional de Aedes aegypti. Esta construção apresenta esterilidade condicionada à presença ou ausência de antibiótico no meio em que esses mosquitos se desenvolvem durante a fase larval. Dessa forma, sem a necessidade de se utilizar radiação para obter insetos estéreis, é possível melhorar a qualidade dos machos adultos liberados e aumentar a competitividade dos mesmos em competir por fêmeas selvagens e adicionalmente gerar o desejado quadro de supressão populacional. / The increasing number of infection cases by arboviruses, for instance Dengue, Zika and Chikungunya, it is necessary to develop new techniques to fight against these arboviruses. Genetic manipulation allowed the production of genetically modified mosquitoes that are capable of suppressing the wild population or prevent pathogens transmission and avoid disease development. The project proposes to establish lines for population suppression of Aedes aegypti, one of the main vectors of those diseases. This construction presents the conditional sterility due to the presence or absence of antibiotics in the environment where these mosquitoes are developed during the larval stage. Thus, without the need to use radiation for sterile insects, it is possible to improve the quality of the released adult males and enhance the competitiveness thereof in competing with wild females and additionally generate population suppression.

O objetivo deste trabalho foi avaliar resíduos de glyphosate e AMPA, produtividade, teor de óleo, conteúdo de proteína bruta e aminoácidos na soja (Glycine max (L.) Merrill) resistente ao glyphosate (SRG) após a aplicação de formulações deste herbicida. Foram desenvolvidos experimentos em replicata (E1 e E2) em lísimetro sob condições de campo e a campo. O objetivo dos experimentos em lísimetro foi quantificar resíduos de três formulações de 14C-glyphosate na SRG. O cultivar de soja utilizada foi o MSOY 7575RR em delineamento experimental inteiramente casualizado, com quatro repetições para ambos os experimentos. Os tratamentos foram glyphosate (sal de isopropilamina), Roundup Original e Roundup Ready na dose de 1,2 kg e.a ha-1 para as três formulações. A aplicação foi realizada aos 25 DAE. Cada planta recebeu 0,96 mg de equivalente ácido (e. a.) de glyphosate com radioatividade total aplicada de 18,33 kBq. A colheita dos grãos, caules, cascas das vagens, folhas e raízes analisadas ocorreram no estágio de maturação plena (R8). A coleta das folhas tratadas ocorreu entre 40 e 55 DAE. Todos os materiais coletados foram moídos. Alíquotas foram oxidadas e quantificadas por espectrometria de cintilação líquida. Não houve diferença estatística entre as formulações para ambos os experimentos, porém formulações com adjuvantes aumentaram os resíduos de glyphosate. Roundup Ready mostrou tendência ao aumento de resíduos em relação aos demais. O valor máximo de resíduos encontrado nos grãos foi de 1,95 mg kg-1. Os experimentos de campo utilizaram a cultivar de soja MSOY 8000RR em delineamento experimental de parcelas subdivididas em blocos ao acaso, com quatro repetições dos seguintes tratamentos: Rodeo, Roundup Original, Roundup Ready, Roundup WG e Controle para as seguintes parcelas (programas): Única - U20 (aplicação única aos 20 dias após a emergencia DAE na dose de 1,2 kg e.a glyphosate ha-1), Única - U30 (aplicação única aos 30 DAE na dose de 1,2 kg e.a glyphosate ha-1), Seqüencial - S20 (aplicação de 0,96 kg e.a glyphosate ha-1 aos 20 DAE e aplicação seqüencial de 0,72 kg e.a. glyphosate ha-1 aos 35 DAE) e Seqüencial - S30 (aplicação de 0,96 kg e.a glyphosate ha-1 aos 30 DAE e aplicação seqüencial de 0,72 kg e.a glyphosate ha-1 aos 45 DAE). A colheita dos grãos analisados em E1 e E2 ocorreu no estágio R8. As variáveis analisadas em grãos da SRG foram: produtividade, teor de óleo, proteína bruta, aminoácidos, resíduos de glyphosate e AMPA. Não houve diferença significativa entre os tratamentos quanto à produtividade, massa de 100 grãos, teor de óleo, conteúdo de proteína bruta e aminoácidos. Houve metabolismo de glyphosate em AMPA e translocação de ambos para os grãos da SRG, exceto para os tratamentos Roundup Original e Roundup Ready, nos programas U20 e S20 do experimento E2. Os maiores níveis de resíduos de glyphosate nos tratamentos Roundup Original e Roundup Ready em U20 e S20 do E2 são devido à presença de adjuvantes destas formulações e provável produtividade de grãos mais baixa do que a média nacional. O maior nível de resíduos de glyphosate foi 0,28 mg kg-1 / The objective of this work was to determine glyphosate and AMPA residues, yield, oil, crude protein content and amino acids in glyphosate-resistant soybean (Glycine max (L.) Merrill) after glyphosate formulations treatment. Experiments (E1 and E2) were performed in lysimeter under field conditions and under field conditions. Lysimeter experiments under field conditions aimed to quantify residues of three formulations of 14C-glyphosate in glyphosate-resistant soybean to this herbicide (GRS). The GRS cultivar used was MSOY 7575RR in a completely randomized design with four replications each treatment. E2 is the E1 replicated 15 days after E1 planting. The treatments were glyphosate isopropylamine, Roundup Ready and Roundup Original at 1.2 kg ae ha-1 for the three formulations. The application was applied on 25 DAE. Each plant received 0.96 mg of acid equivalent (a.e.) glyphosate applied with 18.33 kBq of total radioactivity. Harvest of grains, stems, pod husks, leaves and roots was done at the stage of full maturity (R8). Treated leaves were collected between 40 and 55 DAE. All analysed material was grounded. Aliquotes were oxidized and quantified by liquid scintillation spectrometry. There were no statistical differences among treatments. However, formulations with adjuvants have increased glyphosate residues in GRS. Roundup Ready treatment had a trend to increase residues. The highest glyphosate residual level found in grains was 1.95 mg kg-1. Field experiments were conducted using MSOY 8000 RR variety. The experimental design was a split plot with programmes as main randomized plots and five treatments as subplots having four replications. The treatments were Rodeo, Roundup Original, Roundup Ready, Roundup WG and Control. Programmes were U20 (unique application of 1.2 kg ha-1 a.e. glyphosate at 20 DAE), U30 (unique application of 1.2 kg ha-1 a.e. glyphosate ha-1 at 30 DAE), S20 (apllication of 0.96 kg ha-1 a.e. glyphosate at 20 dae followed by a sequential application of 0.72 kg ha-1 a.e. glyphosate at 35 DAE) and S30 (apllication of 0.96 kg ha-1 a.e. glyphosate at 30 dae followed by a sequential application of 0.72 kg ha-1 a.e. glyphosate at 45 DAE). The analysed variables were: yield, oil content, crude protein, amino acids and glyphosate and AMPA residues. There was glyphosate metabolism in AMPA and translocation of both to GRS grains. The exceptions were Roundup Original and Roundup Ready treatments of E2 on U20 and S20 programs, most likely due the adjuvants of these formulations and the E2 yield lower than the Brazilian average. The highest residue level found in GRS grains was 0.28 mg kg-1