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An integrated BAC approach to the genomics of cork formation
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Orientador(es)
Resumo(s)
Cork is produced by Quercus suber, an evergreen oak with major economic and environmental importance in Portugal. Suberin is the main component of cork but little is known about the molecular processes underlying suberin biosynthesis.
We screened a Quercus suber bacterial artificial chromosome (BAC) library for the presence of relevant genes for suberin biosynthesis through high-density BAC filters hybridizations and we validated hybridization positive clones by Colony PCR. We analyzed BAC-end sequences (BESs) to characterize the structure and composition of the cork oak genome. And we hybridized BAC DNA with mitotic cork oak chromosomes to obtain a cytogenetic map of Q. suber. Finally, we analyzed expression profiles of candidate genes of cork samples from cork oaks with different cork qualities, using quantitative real-time PCR.
We identified several genes involved in suberin biosynthesis pathways. BESs analysis showed high identity with coding regions and suggest the existence of conserved regions within the Fagacea family. We found strong evidence that the genes Cyp 86A1 and GDSL involved in lipid suberin biosynthesis pathway are closely located in the genome. However, we did not succeed in mapping genes of interest in the cork oak genome and we did not find any relation between the genes relative expression and cork quality
Descrição
Mestrado em Biologia Funcional - Instituto Superior de Agronomia
Palavras-chave
cork suberin BAC library BESs FISH quantitative real time PCR
Contexto Educativo
Citação
Pinto, M.J.A.T. - An integrated BAC approach to the genomics of cork formation. Lisboa: ISA, 2013, 129 p.