Identificação de SNPs (Single Nucleiotide Polymorphisms) no gene colina monooxigenase relacionado ao metabolismo da glicina betaína em Eucalyptus
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2010-12-17
Autores
Martin, Leonardo Curi [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Estresses abióticos, como a seca, podem reduzir significativamente os rendimentos no cultivo de espécies comercialmente importantes, tais como o eucalipto na produção de celulose e papel. Quando submetidas às condições de déficit hídrico, as plantas desenvolvem alguns mecanismos de defesa. As betaínas participam destes mecanismos, sendo seu soluto mais comum a glicina betaína. Esta é uma amina quaternária distribuída extensamente em diversas espécies de plantas superiores, sintetizada em elevadas taxas, tendo como função, manter a turgescência celular. A identificação e estudo de genes relacionados à tolerância à seca são importantes para os programas de melhoramento florestal. Este estudo teve por objetivo identificar SNPs (Single Nucleotide Polymorphisms) no gene colina monooxigenase relacionado ao metabolismo da glicina betaína em Eucalyptus. A sequência do gene em estudo foi encontrada em genomas de plantas modelos através do banco de dados GenBank, sendo as mesmas, utilizadas para a procura de similaridades no banco de dados de ESTs de eucalipto FORESTs – FAPESP. Após a constatação de homologia no banco de dados de eucalipto, foram confeccionados oito pares de “primers” para flanquear essas regiões, sendo estes, avaliados, amplificando fragmentos únicos. Depois de realizado o seqüenciamento do gene colina monooxigenase, foi utilizado a ferramenta BLAST no GenBank, confirmando com sucesso a identidade da sequência. Em seguida, as sequências foram alinhadas e os SNPs encontrados. No total, foram identificados 49 SNPs, sendo 12 em regiões codificantes e 37 em regiões UTRs e íntrons. Somente os SNPs localizados nas regiões codificantes foram utilizados neste trabalho, sendo 83,3% deles possuindo mutações sinônimas e 16,7% não-sinônimas. Em seguida, foi utilizada uma população mais abrangente composta de E. grandis...
Abiotic stress, such as drought, can reduce significantly the yield in the important commercially species crop, such as the eucalyptus in the cellulose and paper. When submitted to the water stress conditions, the plants develop some defense mechanisms. The betaines are part of these mechanisms , being their solute more common to the glycine betaine. This is a quaternary amine extensively distributed in several species of higher plants synthesized in high taxes with the function of maintaining the cellular turgescence. Identifying and studying the genes related to the drought tolerance are important for the forest improvement programs. This work aimed at identifying SNPs (Single Nucleotide Polymorphisms) in choline monooxygenase related to the glycine betaine in Eucalyptus. The gene sequence was found in model plants genomes through the databank GenBank, being used for searching similarities in the databank ESTs of eucalyptus FORESTs – FAPESP. After noticing homology in the eucalyptus databank, eight pairs of primers were made to amplify these regions, being evaluated, amplifying unique fragments. After the choline monooxygenase sequencing was performed, it was used the BLAST in the GenBank, confirming successfully the sequence identity. Then, the sequences were aligned and the SNPs were found. In the total, 49 SNPs were identified, being 12 in coding regions and 37 in UTRs and intron regions. Only the SNPs located in coding regions were used in this work, being 83.3% of the SNPs with synonymous mutation and 16.7% non-synonymous. Following, it was used a wider population made up of de E. grandis, E. Urophylla and the hybrid “Urograndis” to perform the SNPs genotyping, establishing the formation of 18 haplotypes and 16 possible haplotypical combinations which revealed that some genotypes were exclusive for the species E. grandis and E. urophylla. With the objective of decreasing costs... (Complete abstract click electronic access below)
Abiotic stress, such as drought, can reduce significantly the yield in the important commercially species crop, such as the eucalyptus in the cellulose and paper. When submitted to the water stress conditions, the plants develop some defense mechanisms. The betaines are part of these mechanisms , being their solute more common to the glycine betaine. This is a quaternary amine extensively distributed in several species of higher plants synthesized in high taxes with the function of maintaining the cellular turgescence. Identifying and studying the genes related to the drought tolerance are important for the forest improvement programs. This work aimed at identifying SNPs (Single Nucleotide Polymorphisms) in choline monooxygenase related to the glycine betaine in Eucalyptus. The gene sequence was found in model plants genomes through the databank GenBank, being used for searching similarities in the databank ESTs of eucalyptus FORESTs – FAPESP. After noticing homology in the eucalyptus databank, eight pairs of primers were made to amplify these regions, being evaluated, amplifying unique fragments. After the choline monooxygenase sequencing was performed, it was used the BLAST in the GenBank, confirming successfully the sequence identity. Then, the sequences were aligned and the SNPs were found. In the total, 49 SNPs were identified, being 12 in coding regions and 37 in UTRs and intron regions. Only the SNPs located in coding regions were used in this work, being 83.3% of the SNPs with synonymous mutation and 16.7% non-synonymous. Following, it was used a wider population made up of de E. grandis, E. Urophylla and the hybrid “Urograndis” to perform the SNPs genotyping, establishing the formation of 18 haplotypes and 16 possible haplotypical combinations which revealed that some genotypes were exclusive for the species E. grandis and E. urophylla. With the objective of decreasing costs... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Genetica vegetal, Eucalipto, Codigo genetico, Plantas - Melhoramento genetico, Glycine betaine
Como citar
MARTIN, Leonardo Curi. Identificação de SNPs (Single Nucleiotide Polymorphisms) no gene colina monooxigenase relacionado ao metabolismo da glicina betaína em Eucalyptus. 2010. 88 f. Dissertação (mestrado) - Universidade Estadual Paulista, Instituto de Biociências de Botucatu, 2010.