Made available in DSpace on 2014-12-17T15:18:08Z (GMT). No. of bitstreams: 1 DaianeCCF.pdf: 438800 bytes, checksum: c847da48b438b366ac3a9e69d018618a (MD5) Previous issue date: 2008-02-28 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Flowering is a process marked by switch of shoot apical meristem to floral meristem, and it involves a complex regulation by endogenous and environmental factors. Analyses of key flowering genes have been carried out primarily in Arabidopsis thaliana and have provided a foundation for understanding the underlying molecular genetic mechanisms controlling different aspects of floral development. Several homologous have been found in other species, but for crops species such as tomatoes this process is not well known. The aim of this work was to use the genetic natural variation associated to the flowering process and use molecular tools such as subtractive libraries and real time PCR in order to identify and analyze the expression from genes that may be associated to flowering in these two species: L. esculentum cv Micro-Tom and L. pimpinellifolium. Our results showed there were identified many genes related to vegetative and possibly to the flowering process. There were also identified many sequences that were unknown. We ve chosen three genes to analyze the expression by real time PCR. The histone H2A gene gave an expression higher in L. pimpinellifolium, due to this the expression of this gene may be associated to flowering in this specie. It was also analyzed the expression of an unknown gene that might be a key factor of the transition to flowering, also in L. pimpinellifolium. For the elongation factor 1-α expression, the expression results were not informative, so this gene may have a constitutive expression in vegetative and flowering state. The results observed allowed us to identify possible genes that may be related to the flowering process. For further results it will be necessary a better characterization of them. / A flora??o ? controlada por condi??es ambientais e fatores end?genos que se associam numa rede de mecanismos gen?ticos bastante complexos. An?lises de genes chaves da flora??o foram feitas primariamente em Arabidopsis thaliana tem fornecido grande conhecimento sobre mecanismos gen?ticos que controlam diferentes aspectos do desenvolvimento floral. Existem muitos hom?logos descritos em diversas outras esp?cies, inclusive em plantas de interesse agron?mico como o tomateiro. Nesta planta, as varia??es gen?ticas naturais relacionadas com a flora??o podem ter origem em diferentes genes expressos durante o desenvolvimento. Com isso, o objetivo deste trabalho foi de prospectar genes associados ao processo de flora??o utilizando a varia??o gen?tica natural existente entre L. esculentum cv Micro-tom e L. pimpinellifolium, aplicando-se a metodologia de biblioteca subtrativa e an?lise de express?o por qPCR em tempo real. Os resultados obtidos permitiram uma identifica??o de alguns genes que possam ser associados ? flora??o nestas esp?cies. Foram encontrados nas bibliotecas subtrativas genes relacionados tanto com o desenvolvimento vegetativo quanto genes ligados ao desenvolvimento reprodutivo, al?m de muitos genes n?o identificados em bancos de dados. Foram analisadas as express?es de tr?s genes por qPCR. As an?lises sugerem uma prov?vel associa??o do gene da histona H2A com a flora??o em L. pimpinellifolium, al?m de ter sido identificado tamb?m um gene desconhecido que pode ser outro fator chave na transi??o do meristema floral nessa esp?cie. Para os dados de express?o do gene fator de elonga??o 1-α, outro gene resultante da subtra??o de uma biblioteca com mensagens vegetativas, n?o foi encontrada liga??o com a flora??o. Visando-se melhor caracterizar este evento fisiol?gico em tomateiro, devem-se realizar estas an?lises para os outros genes identificados.

Made available in DSpace on 2014-12-17T14:10:23Z (GMT). No. of bitstreams: 1 FrancinaldoLS_DISSERT.pdf: 2544554 bytes, checksum: b9889e7b0f272a12f69ccee6cb6a3d3b (MD5) Previous issue date: 2011-05-16 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Flowering is controlled by several environmental and endogenous factors, usually associated with a complex network of metabolic mechanisms. The gene characterization in Arabidopsis model has provided much information about the genetic and molecular mechanisms that control flowering process. Some of these genes had been found in rice and maize. However, in sugarcane this processe is not well known. It is known that early flowering may reduce its production up to 60% at northeast conditions. Considering the impact of early flowering in sugarcane production, the aim of this work was to make the gene characterization of two cDNAs previously identified in subtractive cDNA libraries: scPKCI and scSHAGGY. The in silico analysis showed that these two cDNAs presented both their sequence and functional catalytic domains conserved. The results of transgenic plants containing the overexpression of the gene cassette scPKCI in sense orientation showed that this construction had a negative influence on the plant development as it was observed a decrease in plant height and leaf size. For the scPKCI overexpression in antisense orientation it was observed change in the number of branches from T1 transgenic plants, whereas transgenic T2 plants showed slow development during germination and initial stages of development. The other cDNA analyzed had homology to SHAGGY protein. The overexpression construct in sense orientation did not shown any effect on development. The only difference observed it was an increase in stigma structure. These results allowed us to propose a model how these two genes may be interact and affect floweringdevelopment. / A flora??o ? controlada por diversos fatores como condi??es ambientais e fatores end?genos que se associam em uma rede de mecanismos bastante complexos. A caracteriza??o funcional de alguns genes realizada no modelo vegetal A. thaliana tem fornecido muitas informa??es a respeito dos mecanismos gen?ticos e bioqu?micos que controlam a flora??o. Existem muitos hom?logos descritos em diversas esp?cies, inclusive em plantas de interesse agron?mico, como: arroz e milho. Em cana-de-a??car pouco se sabe sobre esse processo, embora o estudo nessa cultura seja muito importante, uma vez que a flora??o precoce pode acarretar perdas substanciais no teor de sacarose que podem chegar a at? 40% da produ??o. Com isso, o objetivo deste trabalho foi caracterizar a fun??o de dois cDNAs, identificados anteriormente em bibliotecas subtrativas scPKCI e scSHAGGY, por meio de an?lises in silico e a superexpress?o g?nica nas orienta??es senso e antisenso utilizando plantas de Nicotiana tabacum. Os resultados obtidos com a an?lise in silico permitiram observar que os dois cDNAs encontram-se bem conservados, com dom?nios catal?ticos funcionais. Os resultados das plantas transg?nicas contendo o cassete de superexpress?o do gene scPKCI na orienta??o senso mostrou que esta constru??o influenciou negativamente no desenvolvimento normal de plantas de tabaco transg?nicas acarretando a diminui??o da altura m?dia das plantas e da ?rea foliar. Para superexpress?o de scPKCI em orienta??o antissenso, foi observado altera??es no n?mero de ramifica??es das plantas transg?nicas T1, enquanto que as plantas transg?nicas T2 apresentaram o in?cio do desenvolvimento atrasado. Para o outro cDNA analisado, os resultados obtidos mostraram que a superexpress?o do cDNA scSHAGGY na orienta??o senso n?o alterou o desenvolvimento das plantas, por?m a planta apresentou um aumento no tamanho da estrutura floral do gineceu em 100% das flores analisadas (dez flores em seis plantas). Os nossos resultados juntamente com resultados existentes na literatura com outras plantas permitem propor que os cDNAs scPKCI e scSHAGGY estariam envolvidos no processo de flora??o em cana-de-a??car.

Made available in DSpace on 2014-12-17T14:10:25Z (GMT). No. of bitstreams: 1 ValeskaDSS_DISSERT.pdf: 2384968 bytes, checksum: cf4765fd0b42fd627fcb0ce0d604be1e (MD5) Previous issue date: 2011-10-18 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The flowering is a physiological process that it is vital for plants. This physiological process has been well studied in the plant model Arabidopsis, but in sugarcane this process is not well known. The transition of the shoot apical meristem from vegetative to flowering is a critical factor for plant development. At Brazil northeastern region, the transition to flowering in sugarcane has an important effect as it may reduce up to 60% its production. This is a consequence of the sugar translocation from stalks to the shoot apical meristem which is necessary during the flowering process. Therefore, the aim of this work was to explore and analyze cDNAs previously identified using subtractive cDNA libraries. The results showed that these cDNAs showed differential expression profile in varieties of sugarcane (early x late flowering). The in silico analysis suggested that these cDNAs had homology to calmodulin, NAC transcription factor and phosphatidylinositol, a SEC14, which were described in the literature as having a role in the process of floral development. To better understand the role of the cDNA homologous to calmodulin, tobacco plants were transformed with overexpression cassettes in sense and antissense orientation. Plants overexpressing the cassette in sense orientation did not flowered, while plants overexpressing the cassette in the antissense orientation produced flowers. The data obtained in this study suggested the possible role from CAM sequence, SEC14 and NAC in the induction/floral development pathway in sugarcane, this is the first study in order to analyze these genes in the sugarcane flowering process. / A flora??o ? um processo fisiol?gico que demanda um alto gasto energ?tico, e ? vital para a planta, pois ? dela que depende sua propaga??o gen?tica. Este processo fisiol?gico tem sido bem estudado no modelo vegetal Arabidopsis, mas em cana-dea??car n?o ? muito conhecido. A transi??o do meristema apical no processo de flora??o ? um fator cr?tico no ciclo de desenvolvimento da planta. No nordeste brasileiro, a transi??o para a flora??o na cana-de-a??car tem um efeito dram?tico, pois pode reduzir em at? 60% a produ??o de a??car ou etanol. Isso porque, o florescimento da cana resulta na isoporiza??o do colmo que ? caracterizado pela transloca??o do a??car presente no colmo para os ?pices meristem?ticos com a finalidade de suprir a necessidade energ?tica durante o processo de flora??o. Portanto, o objetivo desse trabalho foi de prospectar e analisar cDNAs de cana-dea??car que possam estar associados ? flora??o utilizando ferramentas de bioinform?tica e de transgenia. Os resultados mostraram que os cDNAs em estudo apontaram um perfil de express?o diferencial em variedades de cana-de-a??car. As an?lises in silico sugeriram que os cDNAs estudados apresentam similaridade para calmodulina, fator de transcri??o NAC e fosfatidilinositol, uma SEC14, as quais foram descritas na literatura como tendo um papel no processo de desenvolvimento floral. Para entender melhor o papel do cDNA hom?logo ? calmodulina, plantas de tabaco foram transformadas com cassetes de superexpress?o nas orienta??es sense e antisense contendo o promotor 35S. Plantas superexpressando o cassete na orienta??o senso n?o floresceram, enquanto que plantas superexpressando o cassete na orienta??o antissense apresentaram o desenvolvimento reprodutivo. Os dados obtidos nesse trabalho apontam para o poss?vel papel de CAM, NAC e SEC14 na via de indu??o/desenvolvimento floral em cana-de-a??car, sendo um dos primeiros trabalhos a se estudar esses genes no processo de flora??o nesse organismo

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-07-21T17:14:40Z No. of bitstreams: 1 AmandaLarissaMarquesDeMedeiros_TESE.pdf: 2401701 bytes, checksum: 6c9f1744f688fa158feed1ba0fad56b1 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-07-22T11:33:09Z (GMT) No. of bitstreams: 1 AmandaLarissaMarquesDeMedeiros_TESE.pdf: 2401701 bytes, checksum: 6c9f1744f688fa158feed1ba0fad56b1 (MD5) / Made available in DSpace on 2016-07-22T11:33:09Z (GMT). No. of bitstreams: 1 AmandaLarissaMarquesDeMedeiros_TESE.pdf: 2401701 bytes, checksum: 6c9f1744f688fa158feed1ba0fad56b1 (MD5) Previous issue date: 2015-10-16 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / A cana-de-a??car ? uma importante cultura para nosso pa?s, que contribui com quase metade de toda a produtividade mundial. Muitos desafios s?o enfrentados pelas plantas, que muitas vezes n?o atingem todo o seu potencial gen?tico devido a presen?a de estresses bi?ticos e abi?ticos. Como consequ?ncia, esses estresses podem gerar esp?cies reativas de oxig?nio, que podem danificar o material gen?tico. Outra consequ?ncia do estresse ? a flora??o precoce, que diminui tamb?m a produtividade da cana-de-a??car. Neste contexto, este trabalho apresenta como objetivos caracterizar os genes scMUTM1 e scMUTM2, duas DNA glicosilases pertencentes ? via de reparo por excis?o de base (BER), al?m de identificar transcritos potencialmente relacionados com estresse e reparo de DNA, em duas variedades com fen?tipo contrastante para flora??o. A caracteriza??o das glicosilases incluiu a constru??o de cassetes de express?o de prote?nas, para posterior purifica??o e uso em ensaios enzim?ticos; constru??o de ?rvore filogen?tica e an?lise de promotor deste gene. Com a reconstru??o filogen?tica de MUTM foi poss?vel observar que h? o agrupamento desta sequ?ncia em um ramo com monocotiled?neas e outro com dicotiled?neas. Isto sugere que a duplica??o desta sequ?ncia pode ter ocorrido depois da diverg?ncia desses dois grupos. Com a an?lise do promotor deste gene, ? sugerido que, provavelmente, o gene ScMUTM1 pode estar sofrendo subfuncionaliza??o, uma vez que a regi?o promotora do gene ScMUTM2 apresenta uma maior quantidade de sequ?ncias regulat?rias relacionadas ao estresse. Por meio das an?lises de express?o g?nica utilizando microarranjos, se observou a superexpress?o de genes relacionados ao estresse de forma marcante em uma das condi??es de an?lise relacionadas com flora??o precoce. / Sugarcane is an important culture for Brazil that holds almost half of all worldwide productivity. Plants face many challenges, because of biotic and abiotic stresses presents in the production field, which could prevent plants from reaching their genetic potential. As consequence, those stresses can generate Reactive Oxygen Species ? ROS ? that can cause damages on DNA. Another consequence of stress is the early-flowering process, which contributes for a reduction on yield. In this context, the aim of this work is to characterize ScMUTM1 and ScMUTM2, two DNA glycosylases belonging to base excision repair pathway; and identify genes potentially related to stress and DNA repair in two sugarcane cultivars with contrasting flowering phenotypes. The characterization of the DNA glycosylases included the construction of vector to over express the recombinant proteins ScMUTM1 and ScMUTM2; they will be used in a near future to purification of these proteins and use in enzymatic assays. It was also made a phylogenetic reconstruction of this gene in plants and analysis of its promoter. With the phylogenetic analysis, it is possible to observe the presence of these genes grouped inside a branch with monocots and another one with dicots. This suggests that the duplication of this gene probably occurred after the separation of these two groups. The analysis of the promotor of MUTM shows of the presence of stress-related regulatory motifs at ScMUTM2 promoter, when compared with ScMUTM1. This may suggests that ScMUTM1 might be suffering sub functionalization process. After the analysis of microarrays data, it is observed an up-regulation from some stress-related genes in one of the conditions analyzed, related to early flowering process.

Submitted by Verena Bastos (verena@uefs.br) on 2015-10-02T14:07:29Z No. of bitstreams: 1 45-mara-rubia-dissertacao-completa-21-03-2013.pdf: 2507364 bytes, checksum: ff7d2243b7ca90f0ef456fa17718207d (MD5) / Made available in DSpace on 2015-10-02T14:07:29Z (GMT). No. of bitstreams: 1 45-mara-rubia-dissertacao-completa-21-03-2013.pdf: 2507364 bytes, checksum: ff7d2243b7ca90f0ef456fa17718207d (MD5) Previous issue date: 2013-03-21 / Was conducted a study of reproductive phenology of three species of Miconia Ruiz & Pav. (Melastomataceae) occurring in the gallery forest Len??is river, in Len??is Municipality (12 ? 27'30 "S and 41 ? 27'56" W), Chapada Diamantina, discussing reproductive strategies found in sympatric species with similar morphology of flowers and fruits. The phenological observations were made monthly for 48 months in individuals of the species Miconia alborufescens Naud.,Miconia holosericea (L.) DC. and Miconia prasina (Sw) DC., being accompanied phenophases bud, flower, immature fruit and ripe fruits. The species exhibited a pattern reproductive annual, seasonal, sequential, varying its duration. The offer of flowers occurred typically the beginning of the rainy season and no overlap between species. The offer of fruits occurred since the mid rainy season and the dry season without overlapping. The circular statistical tests indicated strong seasonality of fenofases. The irregularity of rainfall and prolonged drought occurred in the last year of observation influence at the time of the manifestation of reproductive phenology M. holosericea and M. prasina, but they fit for the supply of the resource was not superimposed, favoring the maintenance of pollinators and dispersers to these associates. / Foi realizado um estudo da fenologia reprodutiva de tr?s esp?cies de Miconia Ruiz & Pav. (Melastomataceae) ocorrentes na floresta ciliar do rio Len??is, no Munic?pio de Len??is (12?27?30?S e 41?27?56?W), Chapada Diamantina, Bahia, Brasil, discutindo as estrat?gias reprodutivas encontradas em esp?cies simp?tricas com morfologia similar de flores e frutos. As observa??es fenol?gicas foram realizadas mensalmente por 48 meses em indiv?duos das esp?cies Miconia alborufescens Naud., Miconia holosericea (L.) DC. e Miconia prasina (Sw) DC., sendo acompanhadas as fenofases de bot?o, flor, fruto imaturo e fruto maduro. As esp?cies exibiram um padr?o reprodutivo anual, sazonal, sequencial, com dura??o variada. A oferta de flores ocorreu tipicamente no in?cio da esta??o chuvosa e sem sobreposi??o entre as esp?cies. A oferta de frutos ocorreu desde a esta??o chuvosa at? meados da esta??o seca e sem sobreposi??o. Os testes de estat?stica circular indicaram forte sazonalidade das fenofases.A irregularidade da precipita??o e a estiagem prolongada ocorrida no ?ltimo na o de observa??o influenciou a fenologia reprodutiva de M. holosericea e M. prasina, por?m as mesmas ajustaram-se fenologicamente para que a oferta do recurso n?o fosse sobreposta, favorecendo a manuten??o da fauna de polinizadores e dispersores a estas associadas.

Made available in DSpace on 2014-12-17T15:18:10Z (GMT). No. of bitstreams: 1 MariaAGD.pdf: 3831855 bytes, checksum: 0abe0a10e359f22bab7ed834fa8e296a (MD5) Previous issue date: 2009-02-26 / Sugarcane is one of the most important products of the world and Brazil is responsible for 25 % of the world production. One problem of this culture at northeast of Brazil is the early flowering. In our laboratory, it has been made before four subtractive libraries using early and late flowering genotypes in order to identify messages related to the flowering process. In this work, two cDNAs were chosen to make in silico analysis and overexpression constructs. Another approach to understand the flowering process in sugarcane was to use proteomic tools. First, the protocol for protein extraction using apical meristem was set up. After that, these proteins were separated on two bidimensional gels. It was possible to observe some difference for some regions of these gels as well as some proteins that can be found in all conditions. The next step, spots will be isolated and sequence on MS spectrometry in order to understand this physiological process in sugarcane / A cana-de-a??car ? uma das mais importantes culturas mundiais e atualmente o Brasil representa um dos maiores produtores de cana-de-a??car no ranque mundial. Sabendo-se da import?ncia da cana-de-a??car nos dias atuais, principalmente em rela??o ao biocombust?vel e do problema causado pela flora??o precoce a esta cultura na regi?o Nordeste, foi realizada uma an?lise in silico de dois cDNAs:, 14-3-3 like protein e Protein kinase C inhibitor-like (PKCI), envolvidos no processo de flora??o da cana-de-a??car, utilizando ferramentas gen?micas. Foi escolhido o cDNA PKCI para a constru??o de cassetes de super-express?o de modo a ser caracterizado o papel deste cDNA no processo de flora??o. Outra abordagem utilizada nesse trabalho foi de analisar prote?nas totais de ?pices meristem?ticos de variedades precoce e tardia em g?is uni e bidimensionais. Os resultados mostraram que existem algumas prote?nas que podem ser caracter?sticas de uma das variedades, e em outras foi observado uma express?o diferencial

Made available in DSpace on 2014-12-17T15:18:12Z (GMT). No. of bitstreams: 1 CristianeMF.pdf: 69057 bytes, checksum: 86b55b192a6c5915d66e542325da022c (MD5) Previous issue date: 2007-02-28 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / The northeastern region is responsible to 14.32% of sugarcane national production. This lowered contribution is due to edaphoclimatic condition. Flowering is a vital process to plant which consumes lots of energy and it culminates in a process called isoporization. This one can give in a decreasing of 60% on alcohol and water production. It may consider that cropped sugarcane has a hibrid with octaploid genome, there are varieties with a flowering standard until of non flowering. Using this natural genetic potential on different croppings of sugarcane, the aim of this work was to understand as this process occurs by the usage of subtractive approaches. The total RNA was extracted using Trizol of peaks of merisematics of croppings with induced flowering and other with late flowering. From this total RNA were built four subtractives libraries (B1- induced early flowering subtracted on late flowering not induced; B2- late flowering not induced subtracted induced early flowering; B3- induced early flowering subtracted of not induced early flowering; B02- not induced early flowering subtracted from induced early flowering) using kits Super Smart cDNA synthesis and BD Clontech kit select cDNA subtraction (Clontech). This material was clone don vector pGEM T-easy(Promega) and changed in competent cells of E.coli DH10B. Given analysis sequence was carried out a program BLASTn against database of NCBI and genome of Arabidopsis thaliana, rice and maize. Clones were grouped in 9 different classes according to function. Some factors already related as couples of flower induction were identified at different libraries. And grouped proteins with cell cycle and it controls were presents, mainly kinases proteins. Related factors to proteic sinthesis, metabolism, defence, cell communication were also given in both libraries .Some identified genes did not show similarity on database or homology with hypothesis function, and it can represents new genes to be deposited in international database. These results offers that some identified on sugarcane, classified as on factors classes, cell cycle and cell communication, trough unknown genes, can be linked with genetic changing to the flowering process found in the northeastern region / A regi?o nordeste ? respons?vel por 14,32% da produ??o nacional de cana-de-a??car. Esta contribui??o reduzida ? devido ?s condi??es edafo-clim?ticas da regi?o. A flora??o ? um processo vital para a planta que consome muita energia e culmina num fen?meno denominado de isoporiza??o do colmo. Esta isoporiza??o pode acarretar numa redu??o de at? 60% na produ??o de ?lcool e a??car. Considerando que a cana-de-a??car cultivada corresponde a um h?brido com o genoma octaploide, existem variedades com um padr?o de florescimento precoce at? um padr?o de n?o florescimento. Utilizando este potencial gen?tico natural presente nas diferentes cultivares de cana-de-a??car na regi?o nordeste, o objetivo do presente trabalho foi de compreender como ocorre esse processo de flora??o por meio da metodologia de bibliotecas subtrativas. O RNA total foi extra?do utilizando o reagente Trizol (Invitrogen) de ?pices meristem?ticos de cultivares com florescimento precoce induzida e n?o induzida e outra com florescimento tardio. A partir deste RNA total foram constru?das as quatro bibliotecas subtrativas (B1 florescimento precoce induzida subtra?da da tardia n?o-induzida; B2 florescimento tardio n?o-induzida subtra?do da precoce induzida; B3 precoce induzida subtra?da da precoce n?o-induzida; B02 precoce n?o-induzida subtra?da da precoce induzida), utilizando os kits Super Smart cDNA syntesis e o kit BD Clontech PCR select cDNA subtraction (Clontech). Este material foi clonado no vetor pGEM T-easy (Promega) e transformados em c?lulas competentes de E. coli DH10B. A an?lise das seq??ncias obtidas foi realizada utilizando o programa BLASTn contra o banco de dados do NCBI e do genoma de Arabidopsis thaliana, arroz e milho. Os clones foram agrupados em 9 diferentes classes de acordo com a fun??o. Alguns fatores de transcri??o j? relatados como participantes da via de indu??o floral foram identificados nas diferentes bibliotecas. Al?m disso, prote?nas associadas com o ciclo celular e seu controle tamb?m estavam presentes, destacando-se nesse grupo as prote?nas quinases. Fatores relacionados ? s?ntese prot?ica, metabolismo, defesa, transporte e comunica??o celular, tamb?m foram obtidos em ambas bibliotecas. Alguns genes identificados nas bibliotecas n?o apresentaram similaridade nos bancos de dados ou apresentaram homologia com genes de fun??o hipot?tica, e podem representar novos genes a serem depositados em bancos de dados internacionais. Estes resultados sugerem que alguns dos genes identificados em cana-de-a??car, classificados tanto na classe de fatores de transcri??o, ciclo celular e comunica??o celular, al?m dos genes n?o conhecidos, possam estar associados com a variabilidade gen?tica para o processo de flora??o encontrada na regi?o nordeste

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-01-27T13:32:41Z No. of bitstreams: 1 RenataKalineSouzaEstevam_TESE.pdf: 2591439 bytes, checksum: 8a5092689ed7b6122d0702466b33f854 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-01-31T13:43:08Z (GMT) No. of bitstreams: 1 RenataKalineSouzaEstevam_TESE.pdf: 2591439 bytes, checksum: 8a5092689ed7b6122d0702466b33f854 (MD5) / Made available in DSpace on 2017-01-31T13:43:08Z (GMT). No. of bitstreams: 1 RenataKalineSouzaEstevam_TESE.pdf: 2591439 bytes, checksum: 8a5092689ed7b6122d0702466b33f854 (MD5) Previous issue date: 2016-05-27 / A flora??o ? um processo vital durante o ciclo de vida das plantas e ? marcado pela convers?o do meristema apical vegetativo em reprodutivo devido a intera??es de fatores internos e externos ? planta. Apesar de amplo conhecimento ter sido gerado nessa ?rea, ainda se conhece muito pouco sobre o processo de flora??o e frutifica??o em tomateiro. Pesquisas anteriores realizadas pelo nosso grupo identificaram o cDNA hom?logo a Prote?na Reprimida por Auxina (ARP) em bibliotecas subtrativas reprodutivas de tomateiro. Existem poucos dados sobre a prote?na ARP na literatura, h? relatos de que o gene ARP est? relacionado com a matura??o do fruto em morango, dorm?ncia da gema lateral em ervilha e matura??o do p?len em tabaco, mas a fun??o da prote?na ARP ainda n?o est? clara. Portanto, o intuito deste trabalho foi compreender o papel da prote?na ARP no desenvolvimento vegetativo e nos processos de flora??o e frutifica??o por meio da perda e/ou do ganho de fun??o deste cDNA em plantas transg?nicas de tomateiro contendo o cassete de superexpress?o em orienta??o senso e antissenso para este cDNA. Dessa forma, foram observadas algumas altera??es fenot?picas e estruturais nas plantas transg?nicas (35S::SlARP antissenso e senso), como flora??o e frutifica??o precoce verificada nas plantas 35S::SlARP antissenso nas gera??es T1 a T4, em rela??o ?s controles (transformada com o plasm?deo vazio e n?o transformada). Na an?lise histol?gica, notaram-se ?vulos maduros nas flores das plantas 35S::SlARP antissenso (gera??es T2 a T4), enquanto que as flores das controles n?o apresentaram ?vulos maduros no mesmo per?odo de tempo. Outra modifica??o observada foi o n?mero superior de gemas laterais desenvolvidas nas plantas 35S::SlARP antissenso nas gera??es T3 e T4 quando comparado ?s plantas 35S::SlARP senso e ?s plantas controles (transformada com o plasm?deo vazio e n?o transformada) . Essas produziram ramos com folhas, flores e frutos. Assim como altera??es estruturais nos pec?olos das plantas 35S::SlARP antissenso, incluindo uma aparente quantidade maior de elementos de vaso (xilema e floema) do que os pec?olos as plantas 35S::SlARP senso e mutantes relacionados ? sinaliza??o da auxina (dgt e entire) e plantas controles. Portanto, estes dados sugerem que a prote?na ARP possa estar envolvida na flora??o, frutifica??o e na dorm?ncia das gemas axilares em tomateiro. / Flowering is a vital process during the plant life cycle and it is characterized by the conversion from the vegetative apical meristem into reproductive meristem due to internal and external factors. Despite the considerable knowledge has been produced in this field, not much is known about the flowering and fruiting process in tomato. Furthermore, previous research from our group has identified a cDNA with homology to AUXIN REPRESSED PROTEIN (ARP) in reproductive subtractive libraries from tomato. In the literature, there are few reports where the ARP gene has been associated to fruit ripening in strawberry, dormancy in pea and pollen maturation in tobacco plants, but it is not clear the ARP protein function yet. Therefore, the aim of this work was to understand the role of ARP protein in vegetative development, flowering and fruit set processes through the loss and/or gain of function using this cDNA in transgenic tomato plants with the overexpression cassette constructs in sense and antisense cDNA orientation. Thus, it was observed some phenotypic and structural modifications in transgenic plants (35S::SlARP antisense and sense) such as early flowering and fruiting that was observed in 35S::SlARP antisense plants from the T1 to T4 progeny when compared to controls plants (with the empty vector and wild type plant). In the histological analysis, it was observed mature ovule in 35S::SlARP antisense flowers (progeny T2 to T4), while for the controls plant it wasn?t observed a mature ovule at the same period of time. The other modification observed was a higher number of lateral buds developed in 35S::SlARP antisense plants (progeny T3 and T4) when compared to 35S::SlARP sense and controls plants. These plants produced branches with leaves, flowers and fruits. Besides, it has been observed some structural changes in the petioles from 35S::SlARP antisense plants, including a high number of vessel elements (xylem and phloem) when compared to 35S::SlARP sense, and to dgt and entire mutants (related to auxin signaling) and controls plants (with empty vector and wild type plant). Therefore, these data suggest that ARP protein may be involved in flowering, fruit set and dormancy of axillary buds in tomato.

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.