Differential Gene Expression Analysis In Ethylene And 1-methylcyclopropene Treated Bananas During Ripening Almeida, F.M.F.1; Lajolo, F.M.1; Purgatto, E.1
1Departamento de Alimentos e Nutrição Experimental, FCF-USP, São Paulo – SP – Brazil
The phenotypic changes during the fruit ripening are associated with tissue transformations that lead to texture, flavor, aroma and color changes, turning the fruit acceptable as food. Such alterations elapse of changes in the pattern of gene expression of enzymes and other regulatory proteins involved in ripening, and the hormones, mainly ethylene, exert a key role in the regulation of these processes.The aim of this work is identifying and sequencing genes associated to ripening of banana, isolated from cDNA-AFLP panels of samples treated with ethylene, 1-MCP and without treatment, and after that, to confirm the differential expression of these genes. The selected fragments of differential expressed genes were isolated, re-amplified through polymerase chain reaction, and purified. The PCR product was cloned into pGEM-T vector, and the fragments were sequenced. The sequences had been compared with the deposited ones in the GenBank (NCBI) for gene identification. The confirmation of differential expression of these genes was made through Reverse Northern Blot. Approximately 55% of the genes presented similarity to known genes. Among these, were identified sequences of regulatory proteins, as transcription factors and transcriptional co-repressors; important genes for ionic homeostasis and metal detoxification; proteases and proteases inhibitors; phosphatases; beyond enzymes that participate on carbohydrate, lipid and secondary metabolism. These genes had been classified in accordance with the ethylene signaling dependence, being possible the identification of independent pathways of this signaling. These results can be useful for the understanding of how these metabolic pathways responsible for climacteric fruits ripening are stimulated and linked to each other. Moreover, the information about the influence of ethylene on the regulation of these processes will be able to collaborate for the improvement of techniques applied in fruit post-harvest treatments.
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