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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
471

Análise numérica de risco climático de ocorrência de requeima na cultura da batata na região central do Rio Grande do Sul / Numerical analysis of climatic risk of late blight occurrence in potato crop in the central region of Rio Grande do Sul

Radons, Sidinei Zwick 10 August 2012 (has links)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Diseases and environment are closely related, especially with regard to air temperature and relative humidity. The weather data tend to follow a daily pattern of temporal variation, which derives mainly from the daily variation of the surface radiation balance. Based on this, this study aimed to make a qualitative and quantitative description of the database of the Automatic Meteorological Station of the Instituto Nacional de Meteorologia (INMET) in Santa Maria, RS, determine the hours of greatest frequency of occurrence of daily extreme air temperatures in Santa Maria, develop models to estimate hourly values of air temperature (T) and relative humidity (RH), based on values measured in "UTC standard times" and on the daily minimum air temperature and to determine the risk of fungal diseases occurrence during the developmental cycle of potato in different planting dates, using the Blitecast forecasting system. Models were fitted to estimate air T and partial vapor pressure of water at different times of the day. RH was indirectly obtained from partial vapor pressure of water and saturation vapor pressure, calculated from estimated T values. The database of Principal Climatological Station of Santa Maria was transformed to an hourly estimated T and RH database and the Blitecast's calculated severity values (CSV) were obtained for 14 planting dates in 43 years. The automatic weather station of Santa Maria, RS, belonging to INMET, shows 90.1% of the days of flawless data records in 10 years, from 2002 to 2011. The minimum daily air temperature in Santa Maria occurs more frequently at 09AM UTC, corresponding to 6AM on local official time, from September to February. From March through August, this time is delayed, and its occurrence to be more frequent at 10 h UTC. The precision of the estimates ranged from 0.7 ºC to 1.5 ºC, with average deviation of 0.65 °C for T and 0.7 % to 1.2 %, with average deviation of up to 4% for RH depending on the time. Potato crops during the Fall growing season require on average 54% more fungicide applications compared to the Spring season. In the Spring growing season the most probable number of sprayings ranged from two to four and from six to seven in the Fall growing season. Potato crops planted in August and March were demanded a lower economic, ecological and farmer health cost in crop protection against late blight than earlier crops (July and February). / Doenças e ambiente estão intimamente relacionados, especialmente no que se refere à temperatura e umidade relativa do ar. Os elementos meteorológicos tendem a seguir um padrão de variação temporal diária, que deriva principalmente da variação diária do balanço de radiação da superfície. Com base nisso, o objetivo desse trabalho foi descrever qualitativa e quantitativamente do banco de dados da Estação Meteorológica Automática do Instituto Nacional de Meteorologia de Santa Maria (INMET), determinar os horários de maior frequência de ocorrência das temperaturas extremas diárias do ar em Santa Maria, desenvolver modelos para estimativa dos valores horários de temperatura (T) e umidade relativa do ar (UR) e determinar o risco de ocorrência de requeima no ciclo de desenvolvimento da batata, em diferentes datas de plantio. Foram ajustados modelos analíticos para estimativa da T e da pressão parcial de vapor no ar nos diferentes horários do dia. A UR foi obtida indiretamente, com base nos valores de pressão parcial de vapor no ar estimados e nos valores de pressão de saturação de vapor no ar, calculados com base na T estimada. O banco de dados da Estação Climatológica Principal de Santa Maria foi transformado em um banco de dados horários estimados de T e UR e foram obtidos os valores de severidade calculada (VSC) pelo sistema Blitecast em 14 datas de plantio da batata em 43 anos agrícolas. A estação meteorológica automática de Santa Maria, RS, pertencente ao INMET, apresenta 90,1% dos dias sem falhas de registros de dados em 10 anos, de 2002 a 2011. A temperatura mínima diária do ar em Santa Maria ocorreu com maior frequência às 09 h UTC, correspondendo às 06 h no horário oficial local, nos meses de setembro a fevereiro. De março até agosto, esse horário é atrasado, sendo a sua ocorrência mais frequente às 10 h UTC. A precisão das estimativas variou de 0,7 oC a 1,5 oC, com desvio médio de até 0,65oC para T do ar e de 0,7 % a 1,2 %, com desvio médio de até 4 %, para UR, dependendo do horário. Cultivos de batata no período de safrinha necessitam, em média, de um número 54 % maior de aplicações de fungicida em relação ao período de safra. Na safra o número mais provável de pulverizações oscila entre duas e quatro e na safrinha entre seis e sete. Plantios de batata realizados em agosto e em março exigiram um menor custo econômico, ecológico e de saúde do produtor na proteção da cultura contra a requeima do que plantios mais precoces (julho e fevereiro).
472

Caracterização do período de instabilidade de órgãos vegetais submetidos à injúria mecânica

Albino, Ana Lúcia Seghessi 10 June 2011 (has links)
Made available in DSpace on 2016-08-17T18:39:38Z (GMT). No. of bitstreams: 1 3709.pdf: 10453405 bytes, checksum: e58384df9cb0940e97fe5a0f8206ade2 (MD5) Previous issue date: 2011-06-10 / Universidade Federal de Minas Gerais / Mechanical injuries are a major cause of post-harvest losses, since they may cause metabolic and physiological changes in fruit and vegetables. However there are few studies that characterize the state of the tissue after injury and during the reaction of the tissue, which was called "Period of Instability (PI). In this work the objective was to characterize the "Period of Instability (PI) of tomatoes (Solanum lycopersicum L.) and kale leaves (Brassica oleracea v. acephala L.) after harvest, injured by puncture with different diameters. So, it was realized a tissues histological study of and plants water status analysis subjected to injury by puncture with diameters of 1.5 and 3.5 mm. Analysis of the structure of kale leaf and tomato injured tissues was performed using techniques of light microscopy during 9 and 24 days after harvest, respectively. The water status was evaluated by firmness of tomatoes, turgor pressure and firmness of the kale leaves. They were examined for 19 days in tomatoes and for seven days in kale leaves. Healthy tissues of tomatoes 'Carmen' showed uniform arrangement until the 16th day after harvest. In the tomato injured tissues PI there was adherence to the dead cell walls on the healthy cells. Puncture injuries with diameters of 1.5 and 3.5 mm did not change the firmness of tomatoes in 'Carmen' stored at 25°C, but the tomatoes firmness decreased after two days of post-harvest. The healthy leaves of kale showed degradation of cellular structures after nine days of harvest. The injured leaves by puncturing the 1.5 and 3.5 mm exhibited PI with physiological response, characterized by the accumulation of mucilage in the damaged region. Firmness and turgor pressure were not altered in the kale leaves injured stored at 5°C. However, healthy and injured leaves had a water recovery from the first to the second day after harvest, when stored in refrigerator at 5°C. After the water recovery, the treatments have a decreased of firmness and turgor pressure, and the injured groups showed variation in the values of firmness between the fourth and seventh day of post-harvest. / As injúrias mecânicas são a maior causa de perdas pós-colheita, uma vez que podem causar alterações metabólicas e fisiológicas em frutos e hortaliças. No entanto são escassos estudos que caracterizam o estado do tecido logo após a lesão e durante a reação do tecido lesionado, ao qual denominamos Período de Instabilidade (PI). Neste trabalho o objetivo foi caracterizar o Período de Instabilidade (PI) de tomates (Solanum lycopersicum L.) e folhas de couve-manteiga (Brassica oleracea v. acephala L.), após a colheita, submetidos à lesão mecânica por punção com diferentes diâmetros. Para isso foi realizada a análise histológica dos tecidos e um estudo do estado hídrico desses vegetais submetidos a lesão por punção com diâmetros de 1,5 e 3,5mm. A análise da estrutura dos tecidos da folha de couve-manteiga e de tomate lesionados foi realizada por meio de técnicas usuais de microscopia de luz durante 9 e 24 dias após a colheita, respectivamente. A avaliação do estado hídrico foi feita pelo estudo da firmeza de tomates, e da pressão de turgescência celular e firmeza das folhas de couve-manteiga. As medidas foram realizadas durante 19 dias para tomates e sete dias para as folhas de couve-manteiga. Os tecidos sadios de tomates Carmen apresentaram disposição uniforme até o 16° dia após a colheita. Durante o PI nos tecidos lesionados de tomates verificou-se a adesão das paredes celulares mortas às células sadias quatro dias após a colheita. As lesões por punção com diâmetros de 1,5 e 3,5 mm não alteram a firmeza em tomates Carmem armazenados a 25°C, no entanto os tomates apresentam decaimento da firmeza dois dias após a colheita. As folhas sadias de couve-manteiga apresentaram degradação tardia das estruturas celulares, após o 9° dia de colheita. As folhas lesionadas por punção de 1,5 e 3,5mm apresentam PI com resposta fisiológica, caracterizada pelo acúmulo de mucilagem na região danificada. A firmeza e a pressão de turgescência também não são alteradas nas folhas de couvemanteiga lesionadas por punção de 1,5 e 3,5mm, e armazenadas a 5°C. Porém, folhas sadias e lesionadas apresentam recuperação hídrica do primeiro para o segundo dia após a colheita, quando armazenadas em refrigerador a 5°C. Após a recuperação hídrica, todos os tratamentos apresentam diminuição gradativa da firmeza e da turgescência celular, sendo que os grupos das lesões mostram oscilações nos valores de firmeza entre o quarto e sétimo dia após a colheita.
473

Pré-melhoramento visando resistência à requeima em tomateiro / Pre-breeding for resistance to late blight in tomato

Laurindo, Bruno Soares 17 July 2013 (has links)
Made available in DSpace on 2015-03-26T13:39:59Z (GMT). No. of bitstreams: 1 texto completo.pdf: 425667 bytes, checksum: 1f8c842faf40449adebf9de5ae50ad48 (MD5) Previous issue date: 2013-07-17 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The tomato production is considered a high risk economic activity, and of great agronomic complexity mainly because of the high number of pathogens that causes diseases in the culture. Among the pathogens of greatest importance, there is the oomycete Phytophthora infestans (Mont.) de Bary, causing late blight, which is considered the most destructive disease of tomato. There are no commercial tomato cultivars resistant and the control of this disease is highly dependent on the use of fungicides. Considering that the tomato is one of the most studied crop in genetic terms, the lack of resistant cultivars in the market reflects the difficulty in working this trait in breeding programs. The objectives with this present study was to select sources of resistance to late blight among accessions of Solanum lycopersicum, and estimate the overall and specific ability of combining six tomato accessions of Solanum lycopersicum selected for resistance to late blight, aiming at the selection of potential genitors for pre-breeding for resistance to this disease. Two experiments were carried out. In the first, twelve accessions of S. lycopersicum, from the Vegetable Germplasm Bank of the Federal University of Viçosa - BGH - UFV (BGH-973, BGH-1025, BGH- 2017, BGH-2093, BGH-2095, BGH-2102, BGH-2117, BGH-2127, BGH-2130, BGH- 2332, BGH-2333 and BGH-2343) were evaluated in a randomized block design with three replications. For the evaluation of the resistance, the plants were inoculated with a mixture of sporangia of P. infestans, in the concentration of 5x103 sporangia mL-1 , collected in different regions of the Zona da Mata Mineira. It was evaluated the percentage of plant tissue affected by the disease, or the percentage of severity, evaluated in the form of area under the disease progress curve (AUDPC). The accessions BGH-2102, BGH-2117, BGH-2127, BGH-2130, BGH-2332 and BGH-2343 obtained average values of AUDPC lower than the resistant controls, and were selected as a source of resistance to late blight. The selected accessions as resistant to late blight were used in a second experiment, where a balanced diallel of half table were realized, and fifteen F1 s hybrids were obtained. The six accessions of S. lycopersicum, BGH- 2102, BGH-2117, BGH-2127, BGH-2130, BGH-2332 and BGH-2343 together with the fifteen F1 s hybrid were evaluated in a randomized block design with three replications. For the evaluation of the resistance, the plants were inoculated with a mixture of sporangia of P. infestans, in the concentration of 5x103 sporangia mL-1 collected in different regions of the Zona da Mata Mineira. The percentage of plant tissue affected by the disease, or the percentage of severity, presented in the form of area under the disease progress curve (AUDPC) were evaluated. The diallelic analysis was carried out according to the model of GRIFFING (1956). Observed existence of additive genetic variability between the genitors and the predominance of additive gene effects involved in determining the AUDPC, besides the existence of bidirectional dominance deviations in the control of the character. The diallel analysis was efficient in selecting genitors aimed at a pre-breeding of resistance to late blight, highlighting the accessions BGH- 2117, BGH-2127 and BGH-2343 with the greatest frequency of favorable alleles and divergent, thus they can be included in crossings aiming the pre-breeding for resistance to late blight. / A tomaticultura é considerada uma atividade de elevado risco econômico e grande complexidade agronômica, principalmente pelo elevado número de patógenos que causam doenças na cultura. Entre os patógenos de maior importância, destaca-se o oomiceto Phytophthora infestans (Mont.) de Bary, causador da requeima ou mela, considerada a doença mais destrutiva do tomateiro. Não existem cultivares comerciais de tomate resistentes, e o controle da doença é altamente dependente do uso de fungicidas de caráter preventivo e/ou curativo. Considerando que o tomateiro é uma das plantas cultivadas mais bem estudadas em termos genéticos, a inexistência de cultivares resistentes no mercado reflete a dificuldade em se trabalhar essa característica nos programas de melhoramento. Os objetivos com o presente estudo foram selecionar fontes de resistência à requeima entre acessos de Solanum lycopersicum, e estimar as capacidades geral e específica de combinação dos acessos de S. lycopersicum selecionados quanto a resistência à requeima, visando selecionar potenciais genitores para o pré-melhoramento da resistência a esta doença. Foram realizados dois experimentos. No primeiro avaliou-se doze acessos de S. lycopersicum, do Banco de Germoplasma de Hortaliças da Universidade Federal de Viçosa BGH UFV (BGH- 973, BGH-1025, BGH-2017, BGH-2093, BGH-2095, BGH-2102, BGH-2117, BGH- 2127, BGH-2130, BGH-2332, BGH-2333 e BGH-2343) no delineamento em blocos casualizados, com três repetições. Para a avaliação da resistência, as plantas foram inoculadas com uma mistura de esporângios de P. infestans, na concentração de 5x103 esporângios mL-1, coletados em diferentes regiões da Zona da Mata Mineira. Avaliou-se a porcentagem do tecido vegetal afetado pela doença, ou seja, a porcentagem de severidade, avaliada sob a forma de área abaixo da curva de progresso da doença (AACPD). Os acessos BGH-2102, BGH-2117, BGH-2127, BGH-2130, BGH-2332 e BGH-2343 obtiveram valores médios de AACPD inferiores aos das testemunhas resistentes, sendo selecionados como fonte de resistência à requeima. Os acessos selecionados como resistentes à requeima foram utilizados em um segundo experimento, onde foi realizado um dialelo balanceado de meia tabela, e obtidos quinze híbridos F1 s. Os seis acessos de S. lycopersicum, BGH-2102, BGH-2117, BGH-2127, BGH-2130, BGH-2332 e BGH-2343 juntamente com os quinze híbridos F1 s foram avaliados em delineamento em blocos casualizados, com três repetições. Para a avaliação da resistência, as plantas foram inoculadas com uma mistura de esporângios de P. infestans, na concentração de 5x103 esporângios mL-1, coletados em diferentes regiões da Zona da Mata Mineira. Avaliou-se a porcentagem do tecido vegetal afetado pela doença, ou seja, a porcentagem de severidade, avaliada sob a forma de área abaixo da curva de progresso da doença (AACPD). Procedeu-se a análise dialélica de acordo com o modelo de GRIFFING (1956). Observou-se existência de variabilidade genética aditiva entre os genitores e predominância de efeitos gênicos aditivos envolvidos na determinação da AACPD, além da existência de desvios de dominância bidirecional no controle do caráter em questão. A análise dialélica foi eficiente na seleção de genitores visando o pré-melhoramento da resistência à requeima, destacando os acessos BGH- 2117, BGH-2127 e BGH-2343 como os de maior frequência de alelos favoráveis e divergentes, por isso devem ser incluídos em cruzamentos visando o pré-melhoramento da resistência à requeima.
474

Caracteriza??o funcional de dois cDNAs hom?logos ? ciclofilina e ? prote?na reprimida por auxina (ARP) identificados em bibliotecas subtrativas a para flora??o em tomateiro

Estevam, Renata Kaline Souza 09 September 2011 (has links)
Made available in DSpace on 2014-12-17T14:03:38Z (GMT). No. of bitstreams: 1 RenataKSE_DISSERT.pdf: 1992793 bytes, checksum: 77e111c6a7208973429d97e0cf1a51eb (MD5) Previous issue date: 2011-09-09 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Flowering is a fundamental process in the life cycle for plant. This process is marked by vegetative to reproductive apical meristem conversion, due to interactions between several factors, both internal and external to plant. Therefore, eight subtractive libraries were constructed using apical meristem induced or not induced for two contrasting species: Solanum lycopersicum cv. Micro-Tom and Solanum pimpinellifolium. Several cDNAs were identified and among these, were selected two cDNAs: one homologous cDNA to cyclophilin (LeCYP1) and the other to Auxin repressed protein (ARP). It has observed that LeCYP1 and ARP genes are important in the developmental process to plants. In silico analysis, were used several databases with the exclusion criterion E-value <1.0x10-15. As a result, conservation was observed for proteins analyzed by means of multiple alignments and the presence of functional domains. Then, overexpression cassettes were constructed for the ARP cDNA in sense and antisense orientations. For this step, it was used the CaMV35S promoter. The cDNA orientation (sense or antisense) in relation to the promoter was determined by restriction enzymes and sequencing. Then, this cassette was transferred to binary vector pZP211 and these cassettes were transferred into Agrobacterium tumefaciens LBA4404. S. lycopersicum cv. Micro-Tom (MT) and MT-Rg1 plants were transformed. In addition, seedlings were subjected to hormone treatments using a synthetic auxin (&#61537;- naphthalene acetic acid) and cyclosporin A (cyclophilin inhibitor) treatments and it was found that the hormone treatment there were changes in development of lateral roots pattern, probably related to decreases in auxin signaling caused by reduction of LeCYP1 in MT-dgt plants while cyclosporin A treatments, there was a slight delay in flowering in cv. MT plants. Furthermore, assay with real-time PCR (RT-qPCR) were done for expression level analysis from LeCYP1 and ARP in order to functionally characterize these sequences in tomato plants. / A flora??o ? fundamental no ciclo de vida de uma planta. Este processo ? marcado pela convers?o do meristema apical vegetativo em reprodutivo, devido a intera??es entre diversos fatores, tanto internos quanto externos ? planta. Diante disso, foram constru?das oito bibliotecas subtrativa utilizando ?pices meristem?ticos induzidos e n?o induzidos para a flora??o de duas esp?cies de tomateiro: Solanum lycopersicum cv Micro-Tom e S. pimpinellifolium. In?meros cDNAs foram identificados e dentre estes, foram selecionados o cDNA hom?logo a ciclofilina (LeCYP1) e a Prote?na Reprimida por Auxina (ARP) para a caracteriza??o in silico e funcional em esp?cie de tomateiro. Tem sido observado em outros sistemas vegetais que os genes LeCYP1 e ARP s?o importantes no processo de desenvolvimento da planta. Na an?lise in silico, foram utilizados diversos bancos de dados, tendo como crit?rio de exclus?o o E-value <1.0x10-15. Uma conserva??o para as prote?nas analisadas foi observada por meio dos alinhamentos m?ltiplos e a presen?a de dom?nios funcionais. Em seguida, foram realizadas constru??es de cassetes de superexpress?o para o cDNA hom?logo ? ARP em orienta??o senso e antissenso. Para esta etapa, foi utilizado o promotor forte CaMV35S presente no vetor intermedi?rio pBC (Stratagene). A orienta??o do cDNA (senso ou antissenso) em rela??o ao promotor foi determinada por meio de enzimas de restri??o e sequenciamento dos potenciais clones. Em seguida, este cassete foi transferido para o vetor bin?rio pZP211 e os clones obtidos foram confirmados pelo padr?o de restri??o utilizando diferentes enzimas. Subsequentemente, estes cassetes foram transferidos para a Agrobacterium tumefaciens LBA4404 e posteriormente transformado em plantas de S. lycopersicum cv. Micro-Tom (MT) e MT-Rg1. Em adi??o, pl?ntulas foram submetidas a tratamentos hormonais utilizando uma auxina sint?tica (?cido &#61537;-naftaleno ac?tico) e com ciclosporina A (inibidor de ciclofilina) e foi constatado que no tratamento hormonal houve altera??es no padr?o de desenvolvimento das ra?zes laterais, provavelmente relacionada ao d?ficit na sinaliza??o da auxina ocasionada pela redu??o de LeCYP1 em plantas MT-dgt, enquanto que no tratamento com ciclosporina A, ocorreu um atraso no florescimento de plantas cv MT. Al?m disso, ensaios de PCR em tempo real (RT-qPCR) foram efetuados para an?lise de express?o dos cDNAs hom?logos a LeCYP1 e ARP de modo a caracterizarmos funcionalmente estas sequ?ncias em plantas de tomateiro.
475

Efeito de prepara??es de produtos naturais (cravo da ?ndia e tomate) e de um produto sint?tico na marca??o de constituintes sang?ineos com tecn?cio-99m e na morfologia de hem?cias isoladas de sangue de ratos wistar

Paoli, Severo de 17 August 2007 (has links)
Made available in DSpace on 2014-12-17T14:13:22Z (GMT). No. of bitstreams: 1 SeveroP.pdf: 317890 bytes, checksum: b308c1b0abe5acf0e146092bb45e65ca (MD5) Previous issue date: 2007-08-17 / There are strong interests in the evaluation of the biological effects of natural and synthetic products. Blood constituents labeled with technetium-99m (99mTc) are used in nuclear medicine. The aim of this work was to study the effects of Clove (Caryophyllus aromaticas L.) and OZE (preparation used in the Health Sciences) on the labeling blood constituents with 99mTc and on the morphologic red blood cells (RBC) and the action of an extract of tomato (TO) on the labeling of blood constituents Blood samples were incubated with clove or OZE or TO, stannous chloride and 99mTc. Plasma (P), blood cells (BC), insoluble fractions (IF) of plasma and blood cells were separated. The radioactivity was counted and percentage of radioactivity (%ATI) to each blood fraction was calculated. The shape and morphometric parameter (perimeter/area ratio) were evaluated in the studies with clove and OZE. Clove extract and OZE altered significantly (p<0.05) the %ATI of blood constituents and the shape of red blood cells. However, clove extract not altered the red blood cells perimeter/area ratio. The tomato extract used at the highest concentrations reduced significantly (p<0.05) the %ATI in IF-P, although this extract did not modify the radiolabeling on BC, neither the radioactivity fixation on IFBC. The results indicate that these chemical compounds would have oxidative/chelating actions / Existe um grande interesse na avalia??o dos efeitos biol?gicos de produtos naturais e sint?ticos. Constituintes sang??neos marcados com tecn?cio-99m (99mTc) s?o usados em medicina nuclear. O objetivo desse trabalho foi estudar os efeitos do Cravo da ?ndia (Caryophyllus aromaticus L.) e do ?xido de Zinco Eugenol (OZE) (prepara??o usada em Ci?ncias da Sa?de) na marca??o de constituintes sang??neos com 99mTc e na morfologia de c?lulas vermelhas do sangue (CVS) e a a??o de um extrato de tomate (Solanum lycopersicum) (TO) na marca??o de constituintes sang??neos. Amostras de sangue foram incubadas com cravo da ?ndia ou OZE ou TO, cloreto estanoso e 99mTc. Plasma (P), c?lulas sangu?neas (CS), fra??es insol?veis (IFP) do plasma e das c?lulas sang??neas (IFC) foram separadas. A radioatividade foi contada e o percentual de radioatividade (%ATI) para cada fra??o sang??nea foi calculada. A forma e o par?metro morfom?trico (rela??o per?metro/?rea) foram avaliados nos estudos com o cravo da ?ndia e OZE. Os extratos de cravo da ?ndia e OZE alteraram significativamente (p<0,05) a %ATI dos constituintes sang??neos e a forma das CVS. Entretanto, o cravo da ?ndia n?o foi capaz de modificar a rela??o per?metro/?rea das CVS. O extrato do tomate usado na sua maior concentra??o reduziu significativamente (p<0,05) a %ATI na IF-P, embora esse extrato n?o tenha modificado a radiomarca??o na CS, nem a fixa??o de radioatividade na IF-CS. Os resultados indicam que esses compostos qu?micos poderiam ter a??es oxidativa e/ou quelante. O estudo tem car?ter multidisciplinar com a participa??o das seguintes ?reas do conhecimento: Radiobiologia, Bot?nica,Fitoterapia e Hematologia
476

Extração e caracterização do amido de lobeira (solanum lycocarpum) / Extraction and characterization of the starch lobeira (solanum lycocarpum)

Pascoal, Aline Mendonça 28 March 2014 (has links)
Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2014-12-18T18:07:39Z No. of bitstreams: 2 Tese - Aline Mendonça Pascoal - 2014.doc: 4428288 bytes, checksum: 759c80688c07f85efa0d5d3663f7dbec (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2014-12-22T10:33:16Z (GMT) No. of bitstreams: 2 Tese - Aline Mendonça Pascoal - 2014.doc: 4428288 bytes, checksum: 759c80688c07f85efa0d5d3663f7dbec (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-12-22T10:33:16Z (GMT). No. of bitstreams: 2 Tese - Aline Mendonça Pascoal - 2014.doc: 4428288 bytes, checksum: 759c80688c07f85efa0d5d3663f7dbec (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-03-28 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / This study was conducted to evaluate the chemical, physical-chemical, biochemical and rheological properties of starch from Solanum lycocarpum. The pulp from Solanum lycocarpum fruits was used as raw material for extraction of starch, resulting in a yield of 51%. The starch granules were heterogeneous in size, presenting a conical appearance, very similar to those from high-amylose cassava starch. The elemental analysis (CHNS) revealed 64.33% carbon, 7.16% hydrogen and 0.80% nitrogen. FT-IR spectroscopy showed the peaks characteristic of polysaccharides and NMR analysis confirmed the presence of the α-anomer of D-glucose. The S. lycocarpum starch was characterized by high value of intrinsic viscosity (3515 mPa s) and estimated molecular weight around 645.69 kDa. Furthermore, this starch was classified as a C-type and high amylose content starch, presenting 34.66% of amylose and 38% crystallinity. Endothermic transition temperatures (To = 61.25°C; Tp = 64.5°C; Tc = 67.5°C), gelatinization temperature (∆T = 6.3°C) and enthalpy changes (∆H= 13.21 J g−1) were accessed by DCS analysis. The thermogravimetric analysis of S. lycocarpum starch showed a typical three-step weight loss pattern. Microscopy revealed significant changes in the granule morphology after thermal treatment. Samples treated at 50°C for 10 min lost 52% of their crystallinity, which was partially recovered after storage for 7 days at 4°C. However, samples treated at 65°C for 10 min became were totally amorphous. This treatment was sufficient to completely disrupt the starch granule, as evidenced by the absence of an endothermic peak in the DSC thermogram. The RVA of S. lycocarpum starch revealed 4440.7 cP peak viscosity, 2660.5 cP breakdown viscosity, 2414.1 cP final viscosity, 834.3 cP setback viscosity, and a pasting temperature of 49.6°C. The low content of resistant starch (10.25%) and high content of digestible starch (89.78%) in S. lycocarpum suggest that this starch may be a good source for the production of hydrolysates, such as glucose syrup and its derivatives. These results make the S. lycocarpum fruit a very promising source of starch for biotechnological applications. / Este trabalho foi realizado para avaliar as propriedades químicas, físico- químicas, bioquímicas e reológicas do amido de Solanum lycocarpum. Polpas dos frutos de Solanum lycocarpum foram utilizadas como matéria-prima para a extração de amido, resultando em um rendimento de 51%. Os grânulos de amido apresentaram tamanho heterogêneo, com aparência cônica, muito semelhante ao amido de mandioca, com alto teor de amilose. A análise elementar (CHNS) revelou 64,33% de carbono, 7,16% de hidrogênio e 0,80% de nitrogênio. A espectroscopia FT-IR revelou picos característicos de polissacarídeos e a análise de RMN confirmou a presença do carbono αanomérico da D-glicose. O amido de S. lycocarpum apresentou elevada viscosidade intrínseca (3515 mPa s) e peso molecular estimado de 645,69 kDa. Além disso, este amido foi classificado como um amido do tipo C, apresentando 34,66% de amilose e 38% de cristalinidade. Através de análises por DCS determinou-se as temperaturas de transição endotérmica (To = 61,25°C; Tp = 64,5°C; Tc = 67,5°C), a variação da temperatura de gelatinização (∆T = 6,3°C) e a variação de entalpia (∆H = 13,21 J g-1). A análise termogravimétrica do amido de S. lycocarpum apresentou um padrão típico de perda de peso de três etapas. A microscopia revelou alterações significativas na morfologia dos grânulos após o tratamento térmico. As amostras tratadas a 50°C, durante 10 minutos, perderam 52% de sua cristalinidade, a qual foi parcialmente recuperada após armazenamento a 4°C, por 7 dias. No entanto, as amostras tratadas a 65°C, por 10 minutos, perderam totalmente sua cristalinidade, apresentando estrutura totalmente amorfa. Este tratamento foi suficiente para romper completamente o grânulo de amido, conforme evidenciado pela ausência do pico endotérmico na curva do DSC desta amostra. O RVA do amido de S. lycocarpum revelou um pico de viscosidade de 4440,7 cP, quebra de viscosidade 2660,5 cP, viscosidade final 2414,1 cP, tendência a retrogradação 834,3 cP e temperatura de pasta de 49,6°C. O baixo teor de amido resistente (10,25%) e o elevado teor de amido digerível (89,78%) sugerem que este amido pode ser uma boa fonte para a produção de hidrolisados, tais como xarope de glicose e seus derivados. Os resultados obtidos neste estudo fazem do fruto de S. lycocarpum uma fonte promissora de amido para aplicações biotecnológicas.
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CARACTERISATION DE LA B-GLYCOSIDASE DE LA BLATTE PERIPLANETA AMERICANA : APPLICATION A LA VALORISATION DES GLYCOALCALOÏDES DE LA POMME DE TERRE EN DECOMPOSITION / CHARACTERIZATION OF BETA-GLUCOSIDASE FROM COCKROACH, PERIPLANETA : AMERICANA APPLICATION TO THE VALORIZATION OF GLYCOALCALOIDS FROM DECAYED POTATOES

Koffi, Grokore yvonne 10 November 2016 (has links)
La pomme de terre produit des glycoalcaloïdes comme la plupart des Solanacées. Deux composés, en particulier, l’α-solanine et l’α-chaconine, sont produits en plus grande quantité lorsque le tubercule est exposé à la lumière et subit des dégradations. Ces molécules sont toxiques et peuvent représenter un danger pour le consommateur et des nuisances pour l’environnement. Dans le cadre de cette thèse, les teneurs en α-solanine et α-chaconine dans la chaire de pomme de terre verdie, en germination ou en décomposition retrouvées sur les marchés d’Abidjan (Côte d'Ivoire) ont été analysées. Les résultats ont montré que la chaire de ces pommes de terre contient des quantités élevées de ces deux composés, dépassant 2 à 5 fois la limite recommandée. Pour des raisons de sécurité sanitaire, ces pommes de terre doivent être proscrites de l’alimentation humaine. En revanche, la teneur élevée en glycoalcaloïdes dans ces pommes de terre représente une source de solanidine, un précurseur pour la synthèse d'hormones et de composés pharmacologiquement actifs, qui mérite d’être exploitée. Dans cet objectif, nous avons développé une méthode chimio-enzymatique simple, comprenant un traitement acide partiel suivi d’une hydrolyse enzymatique par la β-glycosidase de la blatte Periplaneta americana dont le gène a été isolé à partir d’une librairie génomique de cDNA afin de détoxifier ces composés et produire la solanidine. / Potato produce glycoalkaloids as most plants of Solanaceae family. The principal glycoalkaloids, α-chaconine and α-solanine are produced in greater quantities when potato tubers are exposed to light and are subject to deteriorations. These compounds are toxic and can represent a real danger for the consumer and the environment where they are discharged during their degradation. In this work, the estimation of glycoalkaloids in the flesh of different types of decayed potatoes usually found in different market places of Abidjan (Ivory Coast) was evaluated. The results showed that turned green and also sprouting or rotting potato flesh contain high amounts of toxic solanine and chaconine, exceeding by 2 to 5-fold the recommended limit. For safety consideration, these decayed potatoes should be systematically set aside. The accumulation of α-chaconine and solanine in potatoes can be seen as an attractive source of solanidine that is an important precursor for hormone synthesis and some pharmacologically active compounds. To this end, we proposed herein a simple chemo-enzymatic protocol comprising a partial acidic hydrolysis followed by an enzymatic treatment with the β-glycosidase from Periplaneta americana whose gene was isolated from a cDNA genomic library in order to detoxify these compounds and produce solanidine.
478

TRANSPIRAÇÃO E CRESCIMENTO FOLIAR DE CLONES DE BATATA E DE MANDIOCA EM RESPOSTA À FRAÇÃO DE ÁGUA TRANSPIRÁVEL NO SOLO / TRANSPIRATION AND LEAF GROWTH OF POTATO AND CASSAVA CLONES AS A FUNCTION OF FRACTION OF TRANSPIRABLE SOIL WATER

Lago, Isabel 18 February 2011 (has links)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The objective of this study was to determine the response of plant transpiration and leaf growth to the fraction of transpirable soil water (FTSW) in potato and cassava clones. Three experiments were conducted with potato, two of them were in Spring, with plantings on 08/20/2008 (BAT1) and on 10/20/2008 (BAT2), and one experiment in Fall, with planting on 03/25/2009 (BAT3). Two experiments were conducted with cassava, with plantings on 09/11/2009 (MAN1) and on 09/08/2010 (MAN2). The experiments were conducted in 8 liter pots filled with soil inside a plastic house. Two potato clones (Macaca and SMINIA 793101- 3) and two cassava clones (Fécula Branca and Fepagro RS 13) were used. Soil water, represented by the FTSW, transpiration and leaf growth were measured on a daily basis during the period of soil drying. The threshold FTSW that starts to affect transpiration, which is an indicator of stomata closure, was 0.38, 0.47 and 0.28 for the clone Macaca and 0.48, 0.51 and 0.32 for the clone SMINIA793101-3 in BAT1, BAT2 and BAT3, respectively, whereas leaf growth decreased at a greater threshold FTSW for Macaca . These results indicate that the clone SMINIA793101-3 is more tolerant to soil water deficit than Macaca . For cassava, the threshold FTSW for transpiration was 0.45 in the clone Fécula Branca and 0.50 in the clone Fepagro RS 13 . The onset of reduction in leaf growth was at a FTSW of 0.51 in the clone Fécula Branca and 0.49 in the clone Fepagro RS 13 .When data were separated in days with low and high vapor pressure deficit at 3PM, threshold FTSW for transpiration and leaf growth changed in both species and within each species. These results confirm the genetic variability available in each species and are a reminder on the complex genotype and environment relationships that characterize agricultural crops. / O objetivo deste trabalho foi determinar a resposta da planta em termos de transpiração e de crescimento foliar em clones de batata e de mandioca ao conteúdo de água disponível no solo, representado pela fração de água transpirável no solo (FATS). Foram realizados três experimentos com a cultura da batata, sendo dois na primavera (cultivo de safra), com plantios em 20/08/2008 (BAT1) e 20/10/2008 (BAT2) e um no outono (cultivo de safrinha), com plantio em 25/03/2009 (BAT3), e dois experimentos com a cultura da mandioca com plantio em 11/09/2009 (MAN1) e 08/09/2010 (MAN2). Os experimentos foram conduzidos em vasos de 8 litros preenchidos com solo e dispostos no interior de um abrigo plástico telado. Os clones de batata utilizados foram Macaca e SMINIA793101-3 e os clones de mandioca foram Fécula Branca e Fepagro RS 13. A água disponível, representada pela FATS, a transpiração e o crescimento foliar foram medidos diariamente durante o período de imposição da deficiência hídrica em cada experimento. Para a cultura da batata a FATS crítica que começa a afetar a transpiração, indicativo do início do fechamento estomático, foi de 0,38, 0,47 e 0,28 no clone Macaca e 0,48, 0,51 e 0,32 no clone SMINIA793101-3, para os experimentos BAT1, BAT2 e BAT3, respectivamente, enquanto o crescimento foliar começou a ser reduzido com um valor de FATS crítica maior para o clone Macaca, indicando que o clone SMINIA793101-3 é mais tolerante ao déficit hídrico no solo que o clone Macaca. Para a cultura da mandioca, a FATS crítica em que a transpiração começa a ser reduzida foi de 0,45 para o clone Fécula Branca e 0,50 para o clone Fepagro RS 13. O crescimento foliar começou a ser reduzido quando a FATS atingiu valores de 0,51 no clone Fécula Branca e 0,49 no clone Fepagro RS 13. Quando os dados foram separados em dias com baixa e alta demanda evaporativa do ar, representada pelo déficit de pressão de vapor do ar às 15 horas, houve diferença na FATS crítica que começa a afetar a transpiração e o crescimento foliar para as duas espécies e dentro de cada espécie. Estes resultados confirmam a variabilidade genética disponível em cada espécie e são um lembrete da complexa relação entre genótipo e ambiente que caracteriza as culturas agrícolas.
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Effets de la contrainte hydrique, seule ou en interaction avec un pathogène, sur le fonctionnement de la plante et la qualité du fruit de Solanum lycopersicum L., en fonction du génotype / Effects of water stress, only or in interaction with a pathogen, on plant functioning and fruit quality, depending on genetic variation

Ripoll, Julie 21 April 2015 (has links)
En horticulture, les effets négatifs du déficit hydrique (WD) sur le rendement peuvent être en partie compensés par des effets positifs sur la qualité des fruits et la stimulation des défenses de la plante face à d’autres stress. L'objectif de la thèse était d'explorer un certain nombre de pistes permettant d'évaluer les effets positifs et négatifs du WD, et d'appréhender les facteurs de variations. Après avoir établi un état des connaissances actuelles des effets du WD sur la qualité des fruits, plusieurs questions ont été abordées: 1) quels sont les effets de WD appliqués à des stades précis du développement de la plante ou du fruit, 2) existe-t-il une mémoire du stress en cas d’alternance de période de WD et de récupération, 3) les défenses de la plante sont-elles stimulées par un WD lors d'une infection par un pathogène, et enfin 4) quelle est la variabilité génétique associée aux réponses au WD au travers des objectifs précédents. Pour cela, plusieurs expérimentations ont été réalisées en serres et en phytotrons. Les parents de la population MAGIC de la tomate (ayant une forte variabilité allélique) ont été étudiés, ainsi que deux tomates commerciales (pour l’étude de l’interaction WD – Botrytis cinerea). Les mécanismes de réponses induits lors du WD ont été caractérisés aux stades végétatif et reproductif à l’échelle de la plante, puis lors des phases de développement du fruit (division cellulaire, expansion cellulaire et maturation). Les stades végétatif et reproductif se distinguent par les mécanismes de gestion du stress oxydatif engendré par le WD. Les plantes végétatives seraient plus affectées au niveau des photosystèmes II alors que les plantes reproductives modulent plutôt leur besoin en eau et sont plus sensibles à l’approche des photosystèmes I. L’impact du WD sur la qualité des fruits diffère en fonction de leurs phases de développement. De forts effets génotypes ont été observés avec une amélioration de la qualité dite gustative (par rapport aux teneurs en sucres et acides) pour un génotype dont les caractéristiques initiales étaient plus faibles que le second génotype testé. L’alternance de périodes de WD et de récupérations constitue un bon compromis entre économie d’eau et adaptation des plantes (via une bonne gestion du stress oxydatif). Cette alternance de stress a des effets neutres à positifs sur les teneurs en sucres des fruits en fonction des génotypes et des réponses très variables pour les teneurs en acides organiques, caroténoïdes et acide ascorbique en fonction des génotypes. L’interaction entre un WD et B. cinerea génère une diminution du ratio C/N associé à une stimulation des défenses de la plante. Cependant l'interaction favorise le développement du pathogène dans les tiges. Cet effet délétère de l'interaction semble lié aux régulations hormonales mises en jeu avec la stimulation de l'ABA (par le WD) et de la voie SA (par B. cinerea et indirectement par le WD) qui sont antagonistes aux voies JA/ET de réponse aux pathogènes nécrotrophes. Toutefois, un effet positif a été observé pour les infections sur feuilles détachées qui constituent un instantané des défenses de la plante, et ce, uniquement pour les plantes déjà infectées sur tige. Ces résultats suscitent de nouvelles interrogations sur la stimulation des défenses de la plante. Ainsi, le WD permet une augmentation du pool de sucres solubles, sans impact notable sur le rendement, avec des effets variables sur les composés secondaires. Le WD permet également dans une certaine mesure de stimuler les défenses de la plante mais de nouvelles recherches sont nécessaires. Enfin, la variabilité génétique des réponses à l’échelle de la plante et du fruit est importante et interagit avec le WD. / In horticulture, yield losses associated to water deficit (WD) could be compensated by positive effects on fruit quality and stimulation of plant to others stresses. The aim of the thesis was to explore a number of questions to assess the positive and negative effects of the WD, and to understand factors variations after setting a current knowledge of the effects of WD quality fruit. The questions were: 1) to characterize the effects of WD applied at specific stages of development of plant or fruit, 2) to understand the stress memory effect by alternating period of WD and recovery, 3) to test the stimulation of plant defences by WD during an infection by a pathogen, and 4) to study the genetic variability associated with responses to WD through the previous objectives. For this, several experiments were conducted in greenhouses and growth chambers. The parents of the MAGIC population of tomato (with rich allelic variability) were studied, as well as two commercial tomatoes (for the study of the interaction between WD and Botrytis cinerea). Plant response mechanisms induced during a WD were characterized in both plant vegetative and reproductive stages, and then during the fruit development stages (cell division, cell expansion and maturation). The plant vegetative and reproductive stages are distinguished by different mechanisms of oxidative stress management generated by WD. Vegetative plants are more affected at the photosystem II while reproductive plants rather modulate their water needs and responses at the photosystem I. The impact of WD on fruit quality differs according to fruit developmental stages. Strong genotypes effects were observed with an important increase in fruit taste quality (according to the concentrations in sugars and acids) for one genotype, which had poorer initial fruit quality than the second. Alternating periods of WD and recovery is a good compromise between water saving and plant adaptation (through proper management of oxidative stress). This alternation of stress has neutral to positive effects on fruit sugar content according to the genotypes and highly variable responses for contents of organic acids, carotenoids and ascorbic acid according to the genotypes. The interaction between a WD and B. cinerea generates a decrease of C/N ratio that can be associated to a stimulation of the pool of plant defence. However, the interaction between B. Cinerea and this WD promote pathogen development in stems. This deleterious effect of interaction seems attributable to hormonal regulation with the stimulation of the ABA pathway (by WD) and the SA pathway (by B. cinerea), which are antagonistic to the JA / ET pathway involved in response to necrotroph pathogens. However, a positive effect was observed for infections in detached leaves of infected plants, detached leaves represent a snapshot of plant defences and these results raise new questions. Overall, these data provide that WD improve fruit taste (sugars and acids) without yield losses but had variables effects on health promoting compounds. WD could permit the stimulation of plant defence but more research is needed. To finish, the genetic variability induced important variability in plant and fruit responses and interact with WD.
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Exploration du rôle de signalisation des Mitogen-Activated Protein Kinases lors de la fécondation chez les plantes

Mazin, Benjamin Damien 10 1900 (has links)
La reproduction est un événement crucial pour la vie des plantes. Ce processus nécessite la formation du pollen et des ovules. Les cellules germinales vont subir la méiose puis une succession de mitoses, deux pour le pollen et trois pour l’ovule, ce qui va leur permettre d’acquérir leurs structures finales. Une fois formés, ces deux gamètes doivent se rencontrer. Pour cela, le grain de pollen va germer sur le stigmate puis former le tube pollinique. Le tube pollinique en croissance va traverser les différents tissus femelles et ainsi tracter les deux cellules spermatiques jusqu’à l’ovule, permettant la reproduction. Un important réseau de signalisation cellulaire est nécessaire pour permettre ces événements. Les cascades des Mitogen-Activated Protein Kinases (MAPKs) sont l’un des réseaux de signalisation les plus étudiés chez les plantes. Ces kinases sont impliquées dans de très nombreux processus développementaux tels que la formation de l’embryon ou des stomates. Pour autant, leurs rôles restent encore peu caractérisés pendant de la fécondation. Ce projet a pour objectif de mieux comprendre le rôle que jouent les MAPK lors de la formation des gamètes mâles et femelles ainsi que lors de la croissance des tubes polliniques. Plusieurs membres de la superfamille des MAPKs ont été caractérisés pour leurs rôles dans la reproduction sexuée des végétaux. De précédents travaux dans le laboratoire de Daniel P. Matton, ont démontré l’implication de deux MAPK Kinases Kinases (MAP3K), la Solanum chacoense Fertilization-Related Kinase 1 (ScFRK1) et ScFRK2. Ces deux kinases sont nécessaires pour le développement de l’ovule et du pollen chez S. chacoense, une espèce de pomme de terre sauvage diploïde. Dans un premier temps, nous avons étudié la fonction d’une nouvelle ScFRK, la ScFRK3. Ce troisième membre de la classe des FRKs chez S. chacoense, est, elle aussi, impliquée dans le développement des gamétophytes mâles et femelles. Du patron d’expression jusqu’à l’établissement d’une voie de signalisation potentielle, en passant par la caractérisation phénotypique des mutants, plusieurs expériences ont été réalisées dans le but de comprendre le rôle de ScFRK3 lors de la formation des gamètes chez Solanum chacoense. Nous montrons que la ScFRK3 est impliquée dans la formation du pollen ainsi que celui des ovules. Nous avons ensuite poursuivi nos recherches en affinant le phénotypage du mutant de surexpression ScFRK2. En effet, les précédentes études ont permis de montré que la surexpression de ScFRK2 conduit le primordium ovulaire à la formation de structures carpéloïdes. Pour autant, les ensembles des primordius ovulaires ne sont pas devenu des strutures capéloïde. Nous montrons ici que seulement 10 % des ovules dans l'ovaire sont devenu 4 des carpéloïde. Notre étude montre qu’en plus des structures carpéloïdes, un grand nombre d'ovule n'ont pas de sac embryonnaire à l'anthèse ce qui explique le faible nombre de graines par fruit. L’analyse du développement des sacs embryonnaires monte que la surexpression de ScFRK2 entraine l’arrêt au stade mégaspore fonctionnelle. Ce phénotype est similaire à ce qui a pu être observé dans les lignées ARN interférant pour la ScFRK1 et la ScFRK3. De précédentes études faites chez Arabidopsis thaliana semblent montrer que les membres de la superfamille des MAPK ne sont pas essentiels pour la croissance du tube pollinique. Pour comprendre le rôle que jouent les MAPK dans l’élongation du tube pollinique, nous avons utilisé un inhibiteur des MAP Kinase Kinase (MKK), appelé U0126. La présence de cette drogue dans le milieu de croissance des grains de pollen provoque une diminution de la germination et de l’élongation du tube pollinique. L’utilisation de la méthode semi-in vivo montre une perte de la polarisation de la croissance des tubes polliniques causée par l’inhibition des MKK. La présence de l’inhibiteur conduit à la diminution de la quantité de filaments d’actine ainsi qu’à leur désorganisation à l’apex du tube. L’exocytose est aussi affectée par l’inhibition des MKK. Les cascades MAPK sont nécessaires à la croissance polarisée du tube pollinique chez Arabidopsis thaliana. Pour finir, nous avons voulu identifier certains membres de la superfamille des MAPK impliqués dans la croissance du tube pollinique. Nous nous sommes intéressés en premier lieu aux orthologues de la famille ScFRK chez Arabidopsis thaliana. Les AtMAP3K19-20-21 sont les orthologues les plus proches de ScFRK3. Ces AtMAP3K sont exprimées lors du développement des grains de pollen et lors de l’élongation de tube pollinique. L’analyse du pollen des différentes lignées mutantes montre qu’en leur absence, le pollen ne présente aucun problème de développement contrairement à ScFRK3. Par contre, les doubles mutants et le triple mutant pour les AtMAP3K19-20-21 montrent une diminution de la capacité de germination. L’élongation du tube pollinique est affectée lors de la mutation d’au moins une des AtMAP3K. Ces deux études démontrent que les MAPK sont essentielles dans la formation et l’élongation du tube pollinique. / Reproduction is a crucial event for plant life. This process requires the formation of pollen and ovules. The germ cells will undergo meiosis and then a succession of mitosis, two for the pollen and three for the ovule, which will allow them to acquire their final structures. Once formed, these two gametes must meet each other. For this, the pollen grain will germinate on the stigmas to form the pollen tube. The growth of the pollen tube will pass through the different female tissues and thus pull the two sperm cells to the ovum for reproduction. An important cellular signaling network is necessary to allow these events to occur. The Mitogen-Activated Protein Kinase (MAPKs) cascades are one of the most studied signaling networks in plants. These kinases are involved in a wide range of developmental processes such as embryo formation and stomata. However, their roles remain poorly characterized during fertilization. The aim of this project is to better understand the role played by MAPKs during the formation of male and female gametes as well as during the growth of pollen tubes. Several members of the MAPK superfamily have been characterized for their role in the sexual reproduction of plants. Previous work in Daniel P. Matton's laboratory has demonstrated the involvement of two MAPK Kinases (MAP3K), Solanum chacoense Fertilization-Related Kinase 1 (ScFRK1) and ScFRK2. These two kinases are necessary for egg and pollen development in S. chacoense, a diploid wild potato species. In a first step, we studied the functionality of a new ScFRK, ScFRK3. This third member of the class of FRKs in S. chacoense, is also involved in the development of male and female gametophytes. From the expression pattern to the establishment of a potential signaling pathway, through the phenotypic characterization of mutants, several experiments have been performed in order to understand the role of ScFRK3 in the formation of gametes in S. chacoense. We show that ScFRK3 is involved in the formation of pollen as well as that of the embryonic sac. We then continued our research by refining the phenotyping of the overexpression mutant ScFRK2. Indeed, previous studies have shown that ScFRK2 overexpression leads the ovular primordium to the formation of carpeloid structures. However, the sets of ovular primordia have not become capeloid structures. We show here that only 10% of the eggs in the ovary have become carpeloid. Our study shows that in addition to the carpeloid structures, a large number of ova do not have an embryonic sac in the anthesis, which explains the low number of seeds per fruit. The analysis of the development of the embryonic sacs shows that 7 overexpression of ScFRK2 leads to the cessation of the functional megaspore stage. This phenotype is similar to what has been observed in interfering RNA lines reducing expression of ScFRK1 and ScFRK3. Previous studies in Arabidopsis thaliana suggest that members of the MAPK superfamily are not essential for pollen tube growth. To understand the role that MAPKs play in pollen tube elongation, we used a MAP Kinase Kinase (MKK) inhibitor called U0126. The presence of this drug in the growth medium of pollen grains causes a decrease in germination and elongation of the pollen tube. The use of the semi in vivo method shows a loss of polarization of the pollen tube growth caused by the inhibition of MKK. The presence of the inhibitor leads to a decrease in the number of actin filaments and their disorganization at the apex of the tube. Exocytosis is also affected by MKK inhibition. We show in this chapter that MAPK cascades are necessary for polarized pollen tube growth in Arabidopsis thaliana. Finally, we wanted to identify some members of the MAPK superfamily involved in pollen tube growth. We were first interested in the ScFRK family orthologs in Arabidopsis thaliana. AtMAP3K19-20-21 are the closest orthologs to ScFRK3. These AtMAP3K are expressed during the development of pollen grains and during the elongation of the pollen tube. Pollen analysis of the different mutant lines shows that in their absence the pollen does not present any development problems unlike ScFRK3. On the other hand, double mutants and triple mutant for AtMAP3K19-20-21 show a decrease in germination capacity. Pollen tube elongation is affected when at least one of the AtMAP3Ks is mutated. These two studies demonstrate that MAPKs are essential for the formation and elongation of the pollen tube and that AtMAP3K19-20-21 participate in these biological processes.

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