Plantas de Citrus regeneradas de tecidos juvenis demandam um longo período para a análise do fenótipo resultante em flores ou frutos. Este trabalho apresenta o mutante espontâneo de florescimento precoce de laranja doce, denominado \'x11\', como modelo para estudos de genômica funcional de Citrus. As frutas cítricas são ricas em carotenoides, pigmentos que possuem grande valor nutricional, como pró-vitamina A (?- ou ?-caroteno). Estudos de genômica funcional envolvendo genes da via biossintética dos carotenoides terão resultados rapidamente obtidos utilizando-se variedades com florescimento e frutificação precoce, como a laranjeira doce mutante \'x11\'. A laranjeira doce \'Sanguínea-de-Mombuca\' (SM) é uma variedade de polpa vermelha que acumula licopeno na polpa dos frutos, podendo ser considerada uma importante ferramenta no estudo da acumulação de carotenoides nos frutos de laranjeiras. Desta maneira, o objetivo deste estudo foi estabelecer a laranjeira \'x11\' como planta-modelo em experimentos de transformação genética visando estudos de genômica funcional dos genes PSY (fitoeno sintase), PDS (fitoeno desaturase), CRTISO (carotenoide isomerase), LCY-b (licpeno ?-ciclase) e ?-caroteno hidroxilase (HYb), envolvidos na biossíntese de carotenoides e utilizar a laranjeira SM como ferramenta no entendimento da acumulação dos carotenoides nos frutos de Citrus. Plantas de laranjeira \'x11\' foram transformadas com promotor de expressão preferencial para órgãos reprodutivos de Citrus controlando a expressão de gene repórter (gusA) para a análise da tecido especificidade. Foi realizada a otimização do protocolo de transformação genética de segmentos de epicótilo de laranjeira \'x11\' via Agrobacterium. A eficiência de transformação média foi de 18,6%, mas atingiu 29,6% no protocolo otimizado. Das 270 brotações positivas, cinco foram microenxertadas e aclimatizadas. Quatro dessas plantas exibiram as primeiras flores em três meses após o estabelecimento ex vitro, e a outra, dois meses mais tarde, independentemente da época do ano. A coloração histoquímica da atividade do gene gusA foi realizada em segmentos de caule, flores e frutos de plantas transgênicas aclimatizadas, com 5-7 meses de idade, confirmando a expressão constitutiva do gene gusA nesses órgãos. As plantas transformadas com a construção para o teste do promotor de expressão preferencial para órgãos reprodutivos de Citrus continuam em fase de desenvolvimento. Em seguida foi realizada a transformação genética de laranjeira \'x11\' para a superexpressão dos genes PDS e LCY-b1 e para a superexpressão do gene LCY-b1 e HYb em laranjeira SM. Também foi realizada a transformação genética de \'x11\' para o silenciamento do gene CRTISO e HYb e também para o silenciamento do gene CRTISO, em SM. A quantificação da expressão do gene-alvo em tecido foliar foi correlacionada ao número de cópias do cisgene inseridos em cada planta, revelando que o nível de expressão gênica não foi diretamente ligado ao número de cópias inseridas no genoma. Uma planta de laranjeira \'x11\' superexpressando o gene LCY-b1 frutificou e foi realizada a análise do perfil de carotenoides totais em relação ao fruto de uma planta não transformada. Neste evento houve aumento no teor de carotenoides totais e xantofilas, indicando o potencial da manipulação dos genes codificadores de enzimas da via biossíntese de carotenoides para alterações quantitativas e qualitativas destes pigmentos / Considering the perennial developmental phase of Citrus, plants regenerated from juvenile tissues require long period for analysis of the resulting phenotype in flowers or fruits. Herein, we present the spontaneous sweet orange mutant named \'x11\', as a model plant for functional genomics studies in Citrus due to the remarkable feature of early-flowering. Citrus fleshy fruits are rich in carotenoids which are pigments that have great nutritional value, such as pro-vitamin A (?- or ?-carotene). Functional genomics studies involving genes related to carotenoids biosynthesis pathway can be rapid generated by using the early flowering mutant, \'x11\' in sweet orange background. The fleshy fruit from the sweet orange \'Sanguínea-de-Mombuca\' (SM) has a red pulp due to accumulation lycopene that may be considered an important tool to investigate the accumulation of carotenoids in sweet orange fruits. Therefore, the aim of this study was to establish the orange mutant \'x11\'as model plants for genetic transformation and functional genomics analysis of key regulatory genes PSY (phytoene synthase), PDS (phytoene desaturase), CRTISO (carotenoid isomerase), LCY-b1 (licopene ?-cyclase) and ?-carotene hydroxylase (HYb) involved in the biosynthesis of carotenoids and to use the sweet orange SM as a tool in understanding the accumulation of carotenoids in sweet orange fruits. The mutant plants \'x11\' were transformed with gene reporter (gusA) under control of specific promoter for reproductive organs in Citrus for the tissue specificity analysis. We report a procedure for efficient regeneration and transformation using epicotyl segment explants of \'x11\' and Agrobacterium tumefaciens as a proof-of-concept. The average transformation efficiency was 18.6%, but reached 29.6% in the best protocol tested. Among 270 positive shoots, five were micrografted and acclimatized. Four of these plants exhibited the first flowers within three months after ex vitro establishment, and the other, two months later, regardless the period of the year. Transgenic plants harboring specific promoter for reproductive organs in Citrus are still under development. We transformed \'x11\' for overexpression of PDS or LCY-b1 genes, and to overexpress LCY-b1 or HYb in SM sweet orange. \'x11\' was transformed to silence CRTISO or HYb , and CRTISO gene in SM sweet orange. Then, quantification of target gene expression in leaf tissue was performed by correlating this information with the number of copies of cisgene inserted into each plant. According to the results, it is suggested that the level of gene expression is not directly linked to the number of copies inserted into the genome.One transgenic plant of \'x11\' overexpressing LCY-b1 produced flowers and fruits and the carotenoid profile analysis indicated an increased in content of total carotenoids and, especially, xanthophylls, clearly indicating the potential to manipulate expression of genes encoding enzymes of the carotenoid biosynthetic pathway to obtain qualitatively and quantitatively changes of these pigments

O manejo químico de plantas daninhas no sistema de plantio por mudas pré-brotadas de cana-de-açúcar, fundamenta a hipótese de que a cultura esteja mais suscetível à intoxicação pelos herbicidas nas aplicações em pré-plantio, devido as raízes das mudas serem transplantadas em contato direto com a solução do solo tratada com os herbicidas aplicados em pré-emergência. Portanto a pesquisa foi desenvolvido com dois objetivos: (1) avaliar a seletividade do herbicida sulfentrazone quando aplicado em área de plantio de cana-de-açúcar no sistema com mudas pré-brotadas com aplicação no pré-plantio utilizando cinco doses (200, 400, 800, 1600 e 3200 g i.a. ha-1); (2) avaliar a seletividade de doses crescentes de clomazone (270, 540, 1080, 2160, e 4320 g i.a. ha-1), quando aplicado em pré-plantio em mudas pré-brotadas (MPB). Os experimentos foram desenvolvidos na Usina São Martinho, próximo ao município de Motuca (SP), sob condições naturais de clima, no período de outubro de 2015 a setembro de 2016. O delineamento experimental foi em blocos ao acaso, com quatro repetições. Os herbicidas foram aplicados no dia anterior ao plantio das mudas pré-brotadas, onde foi utilizada a variedade CTC 11, que foram transplantadas nas parcelas, constituídas por 6 linhas de 10 m, espaçadas de 1,5 m. Foram feitas avaliações visuais com 15, 30, 60, 90 e 120 dias após a aplicação dos tratamentos (DAA), também utilizando-se uma escala visual com notas entre 0 e 100%. Nas avaliações quantitativas avaliou-se os parâmetros altura de plantas (cm), aos 30, 60, 90 e 120 DAA e perfilhamento das mudas aos 60, 90 e 120 DAA. Em todos os períodos avaliados foi utilizado o medidor de clorofila, marca SPAD. Aos 330 DAA, foi realizada a produtividade (t ha-1) integral das parcelas através do corte manual e posterior pesagem com auxílio de uma célula de carga, e foram coletados feixes com dez canas para realização dos parâmetros qualitativos (Brix, Pol, Pureza e ATR). Em ambos trabalhos, os dados de fitointoxicação e produtividade foram previamente transformados na regressão log logística, respeitando o princípio da normalidade dos dados, identificando a dose seletiva para a cultura. Concluiu-se que os herbicidas sulfentrazone (800 g i.a. ha-1), e clomazone (1080 g i.a. ha-1), que são doses comercialmente utilizadas no plantio convencional através de rebolos de cana-de-açúcar, apresentaram sintomas leves de fitointoxicação nas primeiras avaliações, mas com o crescimento e desenvolvimento das mudas, não apresentaram diferença de altura, perfilhamento e índice SPAD, como também na produtividade e características tecnológicas dos colmos não apresentando diferenças significativas. Portanto os herbicidas sulfentrazone e clomazone foram seletivos às mudas de cana-de-açúcar no sistema MPB quando aplicados em pré-plantio para a CTC 11. / The chemical management of plant pests in the planting system with pre-sprouted sugarcane seedlings substantiates the hypothesis that the crop is more susceptible to toxicity by herbicides in pre-planting applications, due to the roots of the seedlings being transferred in direct contact with the soil solution treated with the herbicides applied in the pre-emergence stage. Therefore, this work was prepared with two goals: (1) to assess the selectivity of the herbicide sulfentrazone when applied to a sugarcane planting area in the pre-sprouted seedlings system with application in the pre-planting stage using five rates (200, 400, 800, 1600 and 3200 g a.i. ha-1); (2) to assess the selectivity of growing rates of clomazone (270, 540, 1080, 2160 and 4320 g a.i. ha-1), when applied in the pre-planting stage for pre-sprouted seedlings. The experiments were developed in São Martinho sugar mills, next to the town of Motuca (SP), under natural weather conditions, in the period from October 2015 to September 2016. The experimental design was done in blocks at random, with four repetitions. The herbicides were applied on the day prior to the planting of the pre-sprouted seedlings, where the CTC 11 strain was used, which seedlings were transferred into parcels comprised of 6 lines of 10 m, spaced by 1.5 m. Visual assessments were made 15, 30, 60, 90 and 120 days after the treatment application (DAA), a visual grading scale between 0 and 100% was also used. In the quantitative assessments, the parameters for height of the plants (cm), at 30, 60, 90 and 120 DAA, and tillering at 60, 90 and 120 DAA, were evaluated. In all assessed periods, a direct reading was performed using the chlorophyll meter, SPAD brand. At 330 DAA, the entire productivity (t ha-1) of the parcels was assessed through the manual cut and later weighing with the aid of a load cell, and strips with ten sugarcanes were collected for the performance of qualitative parameters (Brix, Pol, Purity and ATR). In both works, the phytotoxicity and productivity data were previously converted by the log-logistic regression, observing the principle of data normalization, identifying the selective rates for the crop. It was found that herbicides sulfentrazone (800 g a.i ha-1), and clomazone (1080 g a.i. ha-1), which are rates commercially used in the conventional planting through sugarcane billets, presented slight symptoms of phytotoxicity in the first assessments, but with the growth and development of the seedlings, they did not show a difference in height, tillering, SPAD index, and in the yield; and the technological characteristics of the stalks did not present significant differences. Therefore, the herbicides sulfentrazone and clomazone were selective to the sugarcane seedlings in the pre-sprouted seedlings (MPB) system when applied in the pre-planting stage for CTC 11.

Strong light can cause damage and be lethal for photosynthetic organisms. An increase of thermal dissipation of excess absorbed energy at the level of photosynthetic antenna is one of the processes protecting against deleterious effects of light. In cyanobacteria, a soluble photoactive carotenoid binding protein, Orange Carotenoid Protein (OCP) mediates this process. The photoactivated OCP by interacting with the core of phycobilisome (PB; the major photosynthetic antenna of cyanobacteria) triggers the photoprotective mechanism, which decreases the energy arriving at the reaction centres and PSII fluorescence. The excess energy is dissipated as harmless heat. To regain full PB capacity in low light intensities, theFluorescence Recovery Protein (FRP) is required. FRP accelerates the deactivation of OCP.In this work, I present my input in the understanding of the mechanism underlying the OCPrelated photoprotection. I further characterized the FRP of Synechocystis PCC6803, the model organism in our studies. I established that the Synechocystis FRP is shorter than what it was proposed in Cyanobase and it begins at Met26. Our results also revealed the great importance of a high OCP to FRP ratio for existence of photoprotection. The most remarkable achievement of this thesis is the in vitro reconstitution of the OCPrelated mechanism using isolated OCP, PB and FRP. I demonstrated that light is only needed for OCP photoactivation but OCP binding to PB is light independent. Only the photoactivated OCP is able to bind the PB and quench all its fluorescence. Based on our in vitro experiments we proposed a molecular model of OCP-related photoprotection. The in vitro reconstituted system was applied to examine the importance of a conserved salt bridge (Arg155-Glu244) between the two domains of OCP and showed that this salt bridge stabilises the inactive form of OCP. During photoactivation this salt bridge is broken and Arg155 is involved in the interaction between the OCP and the PB. The site of OCP binding in the core of a PB wasalso investigated with the in vitro reconstituted system. Our results demonstrated that the terminal energy emitters of the PB are not needed and that the first site of fluorescence quenching is an APC trimer emitting at 660 nm. Finally, we characterised the properties of excited states of the carotenoid in the photoactivated OCP showing that one of these states presents a very pronounced charge transfer character that likely has a principal role in energy dissipation. Our results strongly suggested that the OCP not only induces thermal energy dissipation but also acts as the energy dissipator.

This Ph.D. thesis is an exploration of carotenoids by ultrafast, time-resolved absorption spectroscopy to investigate their complicated relaxation processes, means of energy transfer, and dependence on structure. The introduction begins with an overview of carotenoids, intended for the reader to appreciate their importance and their complexity as revealed by decades of research in carotenoid photophysics. To understand the primary concerns of this research field, the reader is guided through basic theory of energetic processes, the experimental method, and methods of analysis. The main body of the text is the Research Chapter, containing four sections, each describing research using varied ultrafast transient absorption spectroscopies on carotenoids in solution and when bound to a host protein. Section 2.1 concerns an equilibration phenomenon in the lowest excited state of the carotenoid fucoxanthin in various solutions and temperatures by a multi-pulse transient absorption method. The same method is applied to fucoxanthin in a host antennae protein of the pennate diatom Phaeodactylum tricornutum to investigate the function of the equilibration in energy transfer to Chlorophyll a in Section 2.2. The next two sections regard the effect of carotenoid structure on its relaxation dynamics. Section 2.3 investigates the effect of the non-conjugated acyloxy group of two fucoxanthin derivatives in various solvents. Here, one of the energetic states involved in the equilibrium mentioned above is seen drastically affected. Lastly, Section 2.4 investigates alloxanthin, a carotenoid with an unusual pair of carbon-carbon triple bonds. Their effect on the conjugation is evaluated based upon the molecules' decay dynamics. A general summary and conclusion is provided at the end.

Native to Europe and Asia, Lythrum salicaria is an invasive element on some other continents. Plants in the invaded areas are more robust, higher, and generally more successful compared to individuals in their original area. Based on my results these characteristics cannot be explained by neither higher photosynthesis, chlorophyll and carotenoid mass, nor specific leaf area.

Native to Europe and Asia, Lythrum salicaria is an invasive element on some other continents. Plants in the invaded areas are more robust, higher, and generally more successful compared to individuals in their original area. Based on my results these characteristics cannot be explained by neither higher photosynthesis, chlorophyll and carotenoid mass, nor specific leaf area.

Plantas de Citrus regeneradas de tecidos juvenis demandam um longo período para a análise do fenótipo resultante em flores ou frutos. Este trabalho apresenta o mutante espontâneo de florescimento precoce de laranja doce, denominado \'x11\', como modelo para estudos de genômica funcional de Citrus. As frutas cítricas são ricas em carotenoides, pigmentos que possuem grande valor nutricional, como pró-vitamina A (?- ou ?-caroteno). Estudos de genômica funcional envolvendo genes da via biossintética dos carotenoides terão resultados rapidamente obtidos utilizando-se variedades com florescimento e frutificação precoce, como a laranjeira doce mutante \'x11\'. A laranjeira doce \'Sanguínea-de-Mombuca\' (SM) é uma variedade de polpa vermelha que acumula licopeno na polpa dos frutos, podendo ser considerada uma importante ferramenta no estudo da acumulação de carotenoides nos frutos de laranjeiras. Desta maneira, o objetivo deste estudo foi estabelecer a laranjeira \'x11\' como planta-modelo em experimentos de transformação genética visando estudos de genômica funcional dos genes PSY (fitoeno sintase), PDS (fitoeno desaturase), CRTISO (carotenoide isomerase), LCY-b (licpeno ?-ciclase) e ?-caroteno hidroxilase (HYb), envolvidos na biossíntese de carotenoides e utilizar a laranjeira SM como ferramenta no entendimento da acumulação dos carotenoides nos frutos de Citrus. Plantas de laranjeira \'x11\' foram transformadas com promotor de expressão preferencial para órgãos reprodutivos de Citrus controlando a expressão de gene repórter (gusA) para a análise da tecido especificidade. Foi realizada a otimização do protocolo de transformação genética de segmentos de epicótilo de laranjeira \'x11\' via Agrobacterium. A eficiência de transformação média foi de 18,6%, mas atingiu 29,6% no protocolo otimizado. Das 270 brotações positivas, cinco foram microenxertadas e aclimatizadas. Quatro dessas plantas exibiram as primeiras flores em três meses após o estabelecimento ex vitro, e a outra, dois meses mais tarde, independentemente da época do ano. A coloração histoquímica da atividade do gene gusA foi realizada em segmentos de caule, flores e frutos de plantas transgênicas aclimatizadas, com 5-7 meses de idade, confirmando a expressão constitutiva do gene gusA nesses órgãos. As plantas transformadas com a construção para o teste do promotor de expressão preferencial para órgãos reprodutivos de Citrus continuam em fase de desenvolvimento. Em seguida foi realizada a transformação genética de laranjeira \'x11\' para a superexpressão dos genes PDS e LCY-b1 e para a superexpressão do gene LCY-b1 e HYb em laranjeira SM. Também foi realizada a transformação genética de \'x11\' para o silenciamento do gene CRTISO e HYb e também para o silenciamento do gene CRTISO, em SM. A quantificação da expressão do gene-alvo em tecido foliar foi correlacionada ao número de cópias do cisgene inseridos em cada planta, revelando que o nível de expressão gênica não foi diretamente ligado ao número de cópias inseridas no genoma. Uma planta de laranjeira \'x11\' superexpressando o gene LCY-b1 frutificou e foi realizada a análise do perfil de carotenoides totais em relação ao fruto de uma planta não transformada. Neste evento houve aumento no teor de carotenoides totais e xantofilas, indicando o potencial da manipulação dos genes codificadores de enzimas da via biossíntese de carotenoides para alterações quantitativas e qualitativas destes pigmentos / Considering the perennial developmental phase of Citrus, plants regenerated from juvenile tissues require long period for analysis of the resulting phenotype in flowers or fruits. Herein, we present the spontaneous sweet orange mutant named \'x11\', as a model plant for functional genomics studies in Citrus due to the remarkable feature of early-flowering. Citrus fleshy fruits are rich in carotenoids which are pigments that have great nutritional value, such as pro-vitamin A (?- or ?-carotene). Functional genomics studies involving genes related to carotenoids biosynthesis pathway can be rapid generated by using the early flowering mutant, \'x11\' in sweet orange background. The fleshy fruit from the sweet orange \'Sanguínea-de-Mombuca\' (SM) has a red pulp due to accumulation lycopene that may be considered an important tool to investigate the accumulation of carotenoids in sweet orange fruits. Therefore, the aim of this study was to establish the orange mutant \'x11\'as model plants for genetic transformation and functional genomics analysis of key regulatory genes PSY (phytoene synthase), PDS (phytoene desaturase), CRTISO (carotenoid isomerase), LCY-b1 (licopene ?-cyclase) and ?-carotene hydroxylase (HYb) involved in the biosynthesis of carotenoids and to use the sweet orange SM as a tool in understanding the accumulation of carotenoids in sweet orange fruits. The mutant plants \'x11\' were transformed with gene reporter (gusA) under control of specific promoter for reproductive organs in Citrus for the tissue specificity analysis. We report a procedure for efficient regeneration and transformation using epicotyl segment explants of \'x11\' and Agrobacterium tumefaciens as a proof-of-concept. The average transformation efficiency was 18.6%, but reached 29.6% in the best protocol tested. Among 270 positive shoots, five were micrografted and acclimatized. Four of these plants exhibited the first flowers within three months after ex vitro establishment, and the other, two months later, regardless the period of the year. Transgenic plants harboring specific promoter for reproductive organs in Citrus are still under development. We transformed \'x11\' for overexpression of PDS or LCY-b1 genes, and to overexpress LCY-b1 or HYb in SM sweet orange. \'x11\' was transformed to silence CRTISO or HYb , and CRTISO gene in SM sweet orange. Then, quantification of target gene expression in leaf tissue was performed by correlating this information with the number of copies of cisgene inserted into each plant. According to the results, it is suggested that the level of gene expression is not directly linked to the number of copies inserted into the genome.One transgenic plant of \'x11\' overexpressing LCY-b1 produced flowers and fruits and the carotenoid profile analysis indicated an increased in content of total carotenoids and, especially, xanthophylls, clearly indicating the potential to manipulate expression of genes encoding enzymes of the carotenoid biosynthetic pathway to obtain qualitatively and quantitatively changes of these pigments

Submitted by Franciele Moreira (francielemoreyra@gmail.com) on 2017-10-27T16:23:31Z No. of bitstreams: 2 Dissertação - Joniel Mendes de Araújo - 2017.pdf: 3123838 bytes, checksum: 85b81eeeeeddff88dbab91146cde5709 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-10-30T10:54:37Z (GMT) No. of bitstreams: 2 Dissertação - Joniel Mendes de Araújo - 2017.pdf: 3123838 bytes, checksum: 85b81eeeeeddff88dbab91146cde5709 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-10-30T10:54:37Z (GMT). No. of bitstreams: 2 Dissertação - Joniel Mendes de Araújo - 2017.pdf: 3123838 bytes, checksum: 85b81eeeeeddff88dbab91146cde5709 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-09-28 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Crohn's disease (CD) and ulcerative colitis (RCU) are the major and most prevalent inflammatory bowel diseases (IBD). The lack of effective treatment makes the search for new drugs to treat IBD extremely important. Bixin, a carotenoid present in the species Bixa orellana L., appears as a possible candidate for presenting several biological activities among them antibacterial, antioxidant and anti-inflammatory. The objective of this study was to evaluate the intestinal anti-inflammatory effect of Bixin in experimental TNBS-induced colitis in rats. For the experimental protocol Wistar rats (n = 10) were divided into 7 groups: Saline group (without colitis), TNBS group (with colitis) and groups treated with Bixina gavage at doses of 1, 3, 6, 9 and 12mg / kg. Colitis was induced according to the procedure described by Morris et al. (1989). Euthanasia was performed by CO2 overdose followed by removal of the colon for macroscopic and histological analysis. The results demonstrated that the presence of adhesions and diarrhea was decreased in the groups treated with Bixina. The macroscopic analysis of the colon evidenced the presence of severe damage to the colonic mucosa, with edema, deep ulcerations and hemorrhage in the TNBS group. The Bixina-treated group (12 mg/kg) significantly attenuated the areas of ulcerative lesion and macroscopic score when compared to the TNBS group. Animals receiving Bixin (12 mg/kg) also had a decreased mass / length ratio of the colon compared to the TNBS group. The histological characteristics of the colon of the TNBS group showed severe signs of inflammation, necrosis and edema. However, the Bixin group (12 mg/kg) showed better cell organization and less intestinal epithelial damage when compared to the TNBS group. The results suggest a protective effect of the carotenoid Bixin against experimental colitis induced by TNBS. However, new studies must be performed to verify the mechanisms involved in this activity. / A Doença de Crohn (DC) e a Retocolite Ulcerativa (RCU) são as principais e mais predominantes doenças inflamatórias intestinais (DII). A inexistência de um tratamento eficaz, faz extremamente importante a busca por novos fármacos para tratar as DII. A Bixina, um carotenoide presente na espécie Bixa orellana L., aparece, como um possível candidato por apresentar diversas atividades biológicas dentre elas antibacteriana, antioxidante e anti-inflamatória. O objetivo deste trabalho foi avaliar o efeito anti-inflamatório intestinal da Bixina na colite experimental induzida por TNBS em ratos. Para a realização do protocolo experimental utilizou-se fêmeas de ratos Wistar (n = 10) divididas em 7 grupos: grupo Salina (sem colite), grupo TNBS (com colite) e grupos tratados por gavagem com Bixina nas doses de 1, 3, 6, 9 e 12mg/Kg. A colite foi induzida de acordo com o procedimento descrito por Morris et al. (1989). E a eutanásia foi realizada por overdose de CO2, seguida da remoção do cólon para realização das análises macroscópicas e histológicas. Os resultados demonstraram que a presença de adesões e diarreia, foi diminuída nos grupos tratados com Bixina. A análise macroscópica do cólon evidenciou a presença de danos graves na mucosa cólica, com edema, ulcerações profundas e hemorragia no grupo TNBS. O grupo tratado com Bixina (12 mg/kg) atenuou significativamente as áreas de lesão ulcerativa e o escore macroscópico quando comparado ao grupo TNBS. Os Animais que receberam Bixina (12 mg/kg) apresentaram também uma relação massa/comprimento do cólon diminuída em comparação com o grupo TNBS. As características histológicas do cólon de ratos do grupo TNBS apresentaram sinais severos de inflamação, necrose e edema. Contudo, o grupo Bixina (12 mg/Kg), apresentou uma melhor organização celular e menos danos do epitélio intestinal quando comparado ao grupo TNBS. Os resultados sugerem um efeito protetor do carotenoide Bixina contra a colite experimental induzida pelo TNBS. Contudo, novos estudos devem ser realizados para verificar os mecanismos envolvidos nessa atividade.

O manejo químico de plantas daninhas no sistema de plantio por mudas pré-brotadas de cana-de-açúcar, fundamenta a hipótese de que a cultura esteja mais suscetível à intoxicação pelos herbicidas nas aplicações em pré-plantio, devido as raízes das mudas serem transplantadas em contato direto com a solução do solo tratada com os herbicidas aplicados em pré-emergência. Portanto a pesquisa foi desenvolvido com dois objetivos: (1) avaliar a seletividade do herbicida sulfentrazone quando aplicado em área de plantio de cana-de-açúcar no sistema com mudas pré-brotadas com aplicação no pré-plantio utilizando cinco doses (200, 400, 800, 1600 e 3200 g i.a. ha-1); (2) avaliar a seletividade de doses crescentes de clomazone (270, 540, 1080, 2160, e 4320 g i.a. ha-1), quando aplicado em pré-plantio em mudas pré-brotadas (MPB). Os experimentos foram desenvolvidos na Usina São Martinho, próximo ao município de Motuca (SP), sob condições naturais de clima, no período de outubro de 2015 a setembro de 2016. O delineamento experimental foi em blocos ao acaso, com quatro repetições. Os herbicidas foram aplicados no dia anterior ao plantio das mudas pré-brotadas, onde foi utilizada a variedade CTC 11, que foram transplantadas nas parcelas, constituídas por 6 linhas de 10 m, espaçadas de 1,5 m. Foram feitas avaliações visuais com 15, 30, 60, 90 e 120 dias após a aplicação dos tratamentos (DAA), também utilizando-se uma escala visual com notas entre 0 e 100%. Nas avaliações quantitativas avaliou-se os parâmetros altura de plantas (cm), aos 30, 60, 90 e 120 DAA e perfilhamento das mudas aos 60, 90 e 120 DAA. Em todos os períodos avaliados foi utilizado o medidor de clorofila, marca SPAD. Aos 330 DAA, foi realizada a produtividade (t ha-1) integral das parcelas através do corte manual e posterior pesagem com auxílio de uma célula de carga, e foram coletados feixes com dez canas para realização dos parâmetros qualitativos (Brix, Pol, Pureza e ATR). Em ambos trabalhos, os dados de fitointoxicação e produtividade foram previamente transformados na regressão log logística, respeitando o princípio da normalidade dos dados, identificando a dose seletiva para a cultura. Concluiu-se que os herbicidas sulfentrazone (800 g i.a. ha-1), e clomazone (1080 g i.a. ha-1), que são doses comercialmente utilizadas no plantio convencional através de rebolos de cana-de-açúcar, apresentaram sintomas leves de fitointoxicação nas primeiras avaliações, mas com o crescimento e desenvolvimento das mudas, não apresentaram diferença de altura, perfilhamento e índice SPAD, como também na produtividade e características tecnológicas dos colmos não apresentando diferenças significativas. Portanto os herbicidas sulfentrazone e clomazone foram seletivos às mudas de cana-de-açúcar no sistema MPB quando aplicados em pré-plantio para a CTC 11. / The chemical management of plant pests in the planting system with pre-sprouted sugarcane seedlings substantiates the hypothesis that the crop is more susceptible to toxicity by herbicides in pre-planting applications, due to the roots of the seedlings being transferred in direct contact with the soil solution treated with the herbicides applied in the pre-emergence stage. Therefore, this work was prepared with two goals: (1) to assess the selectivity of the herbicide sulfentrazone when applied to a sugarcane planting area in the pre-sprouted seedlings system with application in the pre-planting stage using five rates (200, 400, 800, 1600 and 3200 g a.i. ha-1); (2) to assess the selectivity of growing rates of clomazone (270, 540, 1080, 2160 and 4320 g a.i. ha-1), when applied in the pre-planting stage for pre-sprouted seedlings. The experiments were developed in São Martinho sugar mills, next to the town of Motuca (SP), under natural weather conditions, in the period from October 2015 to September 2016. The experimental design was done in blocks at random, with four repetitions. The herbicides were applied on the day prior to the planting of the pre-sprouted seedlings, where the CTC 11 strain was used, which seedlings were transferred into parcels comprised of 6 lines of 10 m, spaced by 1.5 m. Visual assessments were made 15, 30, 60, 90 and 120 days after the treatment application (DAA), a visual grading scale between 0 and 100% was also used. In the quantitative assessments, the parameters for height of the plants (cm), at 30, 60, 90 and 120 DAA, and tillering at 60, 90 and 120 DAA, were evaluated. In all assessed periods, a direct reading was performed using the chlorophyll meter, SPAD brand. At 330 DAA, the entire productivity (t ha-1) of the parcels was assessed through the manual cut and later weighing with the aid of a load cell, and strips with ten sugarcanes were collected for the performance of qualitative parameters (Brix, Pol, Purity and ATR). In both works, the phytotoxicity and productivity data were previously converted by the log-logistic regression, observing the principle of data normalization, identifying the selective rates for the crop. It was found that herbicides sulfentrazone (800 g a.i ha-1), and clomazone (1080 g a.i. ha-1), which are rates commercially used in the conventional planting through sugarcane billets, presented slight symptoms of phytotoxicity in the first assessments, but with the growth and development of the seedlings, they did not show a difference in height, tillering, SPAD index, and in the yield; and the technological characteristics of the stalks did not present significant differences. Therefore, the herbicides sulfentrazone and clomazone were selective to the sugarcane seedlings in the pre-sprouted seedlings (MPB) system when applied in the pre-planting stage for CTC 11.

Les interactions entre l’auxine et l’éthylène sont complexes et contrôlent divers processus de développement des plantes tels que l’élongation racinaire ou la différentiation des racines secondaires. Mais, il existe peu d’études montrant le rôle des interactions entre ces deux hormones au cours du développement et de la maturation des fruits. Le changement de couleur des fruits chez la tomate est une caractéristique de la maturation qui est associée à la fois à la dégradation des chlorophylles et à l’accumulation des caroténoïdes. Dans ce travail, l’application exogène d’auxine et d’éthylène a montré l’impact de ces deux hormones sur la maturation de la tomate et en particulier sur le changement de couleur des fruits. Nous avons montré que l’acide indol-acétique (IAA) retarde la transition du vert à l’orange/rouge, alors que l’éthylène, apporté sous la forme d’acide 1-aminocyclopropane-1-carboxylique (ACC), son précurseur, accélère la coloration des fruits. Par contre, l’inhibition de l’auxine par le PCIB, un antagoniste de l’auxine, provoque les mêmes effets que l’éthylène. L’analyse des caroténoïdes montre que l’ACC comme le PCIB augmente la teneur en lycopène et diminue la teneur en carotène alors que l’IAA provoque l’effet inverse. L’étude de l’accumulation des ARNs messagers de plusieurs gènes clés de la voie de biosynthèse des caroténoïdes a montré que le gène β-lcy codant pour la lycopène cyclase joue un rôle clé dans le contrôle de la biosynthèse et de l’accumulation des pigments et que son expression est fortement dépendante de l’équilibre auxine-éthylène. D’autre part, nos résultats ont montré que le gène rin joue un rôle important dans le contrôle de l’expression des gènes clés de la voie de biosynthèse des caroténoïdes. Pour avoir une meilleure vision des gènes différentiellement exprimés par l’auxine et l’éthylène au cours de la maturation, l’analyse du transcriptome des fruits traités par de l’ACC et de l’IAA a été réalisée par RNA-Seq au laboratoire. Parmi les facteurs de transcriptions étudiés, le gène Sl-IAA17, un membre de la famille des AUX/IAA, est fortement affecté par l’auxine et l’éthylène. La caractérisation fonctionnelle du gène Sl-IAA17 pendant le développement du fruit a été réalisée en créant des lignées transgéniques sous exprimant ce gène en mettant en œuvre la stratégie des ARNs interférents. Ces lignées présentent un phénotype caractéristique produisant des fruits de plus gros calibre que celui des fruits sauvages. Les analyses histologiques des tissus des fruits ont montré que ce phénotype est associé à un péricarpe plus épais. En microscopie, nous avons constaté que l’augmentation de l’épaisseur du péricarpe dans les lignées transgéniques n’était pas due à un plus grand nombre de cellules mais à l’augmentation de la taille des cellules. Enfin, nous avons observé que l’expansion des cellules dans les fruits transgéniques est étroitement couplée avec des niveaux de ploïdie plus élevés que dans les fruits sauvages, ce qui suggère une stimulation du processus endoréduplication. Ces résultats démontrent très clairement l’existence d’une étroite relation entre la signalisation de l’auxine, le contrôle de la taille du volume cellulaire et le processus d’endoréduplication. En conclusion, les résultats présentés fournissent des connaissances nouvelles sur les interactions entre l’auxine et l’éthylène au cours du développement du fruit et en particulier au cours de la transition fruit immature - fruit mature. De plus, ils apportent des éléments nouveaux sur la connaissance du rôle de la voie de signalisation de l’auxine dans le contrôle du développement des fruits charnus et en particulier sur la fonction de certains membres des AUX/IAA sur la détermination du volume et du poids des fruits. / The interaction between auxin and ethylene are complex and control various processes of plant development, such as root elongation or differentiation of secondary roots. But there are few studies showing the role of interactions between these two hormones during development and maturation of the fruit. The color change in the tomato fruit is a feature of the maturation that is associated with the degradation of the chlorophyll and carotenoid accumulation. In this work, the application of exogenous auxin and ethylene showed the impact of these two hormones in the tomato ripening and in particular the change of fruit color. We have shown that indole-acetic acid (IAA) delays the transition from green to orange / red, while ethylene, supplied as 1-aminocyclopropane-1-carboxylic acid form (ACC), its precursor, accelerated this transition. However the auxin inhibition by p-chlorophenoxy isobutyic acid (PCIB), an auxin antagonist, caused the same effects similar to ethylene. The carotenoid analysis showed that the ACC and PCIB increase the lycopene content and reduced the carotene content while IAA causes the opposite effect. The study of the accumulation of mRNAs for several key genes of the carotenoid biosynthetic pathway has shown that the gene β-lcy encoding lycopene cyclase plays a key role in the control of biosynthesis and accumulation of pigments and that its expression is highly dependent on the auxin-ethylene balance. In addition, our results showed that the rin gene plays an important role in controlling the expression of the key carotenoid biosynthetic pathway genes. To get a better view of differentially expressed genes by auxin and ethylene during ripening, transcriptome analysis of fruits treated with ACC and IAA was performed by a preliminary RNA-Seq approach. Among the transcription factors studied in the laboratory, the gene Sl-IAA17, a member of the family of Aux/IAA was affected by auxin and ethylene. Functional characterization of Sl-IAA17 gene during fruit development was performed by creating transgenic lines under-expressing this gene by RNAi. These lines display a phenotype producing bigger fruit than wild type. Histological analysis of the tissues showed that fruit phenotype is associated with a thicker pericarp. By microscopy, we observed that increasing the thickness of the pericarp in the transgenic lines was not due to a greater number of cells but to the increase in cell size. Finally, we observed that cell expansion in transgenic fruit is tightly coupled with higher ploidy levels than wild fruits, suggesting a stimulation of the endoreduplication process. These results clearly demonstrate the existence of a close relationship between the auxin signal, the control cell size, fruit volume and the endoreduplication process. In conclusion, the results provide new insights into the interactions between auxin and ethylene during fruit development and in particular during the transition immature fruit, mature fruit. In addition, they provide new information on the understanding of the role of the signaling pathway of auxin in controlling the development of fleshy fruits and in particular on the basis of certain members of the AUX/IAA on regulating volume and fruit weight.