Análise dos padrões de metilação do DNA de feijoeiro em resposta ao déficit hídrico
Drought is an abiotic stress which causes great losses in productivity. In response to stress are triggered behavioral, morphological, anatomical, physiological and biochemical changes which are dependent on molecular signaling processes including transcription factors. Among the molecular processes...
| Autor principal: | Kovali, Kamila Katieli |
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| Formato: | Trabalho de Conclusão de Curso (Graduação) |
| Idioma: | Português |
| Publicado em: |
Universidade Tecnológica Federal do Paraná
2020
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| Assuntos: | |
| Acesso em linha: |
http://repositorio.utfpr.edu.br/jspui/handle/1/14094 |
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| Resumo: |
Drought is an abiotic stress which causes great losses in productivity. In response to stress are triggered behavioral, morphological, anatomical, physiological and biochemical changes which are dependent on molecular signaling processes including transcription factors. Among the molecular processes can be highlighted epigenetic changes, responsible for heritable changes in gene expression without alterations in gene sequence. DNA methylation is an epigenetic change, and the identification of methylated cytosines in DNA can be accomplished by using restriction enzymes sensitive to methylation (RESM) associated with the use of PCR (Reaction Polimerse DNA chain). The objective of this study was to analyze the changes in the common bean DNA methylation pattern in response to water deficit. The experimental design was randomized blocks with factorial arrangement and three replications, four cultivars (IPR Curió, ANFC 9, Pérola and IAC Imperador and four drought levels (100 - control, 75, 50 and 25% of capacity field). Irrigation was suppressed when the plants reached the V2 stage. DNA was extracted from leaves treated with ERSM HpyCH4IV, amplified by PCR and visualized by agarose gel electrophoresis. We tested seven pairs of primers flanking the restriction site located in the gene and promoter DNA sequences. Changes in methylation patterns were observed in promoters of genes studied. 50% and 75% of field capacity were no differences in the methylation pattern presented in the control in all genotypes. When soil moisture reached 25% of field capacity, IPR Curió genotype demethylated DNA in the promoter region from Phvul.001G187100 locus that corresponds to the transcription factor APETALA2 / ETHYLENE RESPONSE FACTOR (AP2 / ERF), while in the same sequence, IAC Imperador kept the methylation which was also observed in the control condition, in the other genotypes was not observed demethylation in any condition for this promoter. In the promoter of the Phvul.005G126300 locus which also belongs to the AP2 / ERF family, demethylation was observed in IAC Imperador in 25% of field capacity, the other genotypes remained methylated in all analyzed conditions. These changes may indicate possible changes in gene expression in order to increase the plasticity of genotypes face to stress. |
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