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Prospec??o e caracteriza??o de genes associados ao processo de indu??o floral em cana-de-a??carFurtado, Cristiane Miranda 28 February 2007 (has links)
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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
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