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Structural and phylogenetic characterization of a polyphenol oxidase gene in lulo (Solanum quitoense Lam.)
Caracterización estructural y filogenética de un gen de polifenol oxidasa en lulo (Solanum quitoense Lam.)
DOI:
https://doi.org/10.15446/agron.colomb.v43n1.118599Keywords:
Solanaceae, catechol oxidase, browning, phenolic compounds, protein domain, naranjilla (en)Solanaceae, catecol oxidasa, pardeamiento, compuestos fenólicos, dominio proteico, naranjilla (es)
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The lulo or naranjilla (Solanum quitoense Lam.) is one of the most important Colombian native fruits. The sale and industrial processing of fresh fruit is severely limited by enzymatic browning. Until now, there was no knowledge about polyphenol oxidases (PPO) in lulo. The aim of this study was to understand some structural and phylogenetic aspects of the first lulo ppo gene that has been characterized. Using two pairs of degenerate primers, two fragments of lulo genomic DNA were isolated by PCR, sequenced and assembled into a partial sequence of 1417 bp (SquPPO1) lacking introns. Hybridization of a 920-bp probe generated from a potato ppo gene with a 12 kb region of BamHI-PstI, BamHI-XbaI and XbaI-PstI digested lulo DNA confirmed the presence of at least one ppo gene in this species. While two conserved sites (Tyr-1 and Tyr-2) have been identified in the copper-binding domains of other Solanaceae PPOs, no Tyr-2 site was found in lulo PPO because of a conservative substitution DxE in this region. Phylogenetic analysis placed the SquPPO1 gene in the same cluster as the SmePPO4, SmePPO5, and SmePPO6 eggplant (Solanum melongena L.) genes. Our results show that SquPPO1 is phylogenetically closer to eggplant ppo genes than to those of potato, tobacco, and tomato and that it exhibits a variation that modifies the distribution of protein-conserved sites. These findings offer new insights into the molecular basis of enzymatic browning in lulo and may inform strategies to reduce postharvest losses.
El lulo o naranjilla (Solanum quitoense Lam.) es una de las frutas nativas colombianas más importantes. La venta y el procesamiento industrial de la fruta fresca son severamente limitados por el pardeamiento enzimático. Hasta ahora, no había conocimiento sobre las polifenol-oxidasas (PPO) en lulo. El objetivo de este estudio fue conocer algunos aspectos estructurales y filogenéticos del primer gen ppo de lulo caracterizado. Utilizando dos pares de iniciadores degenerados, dos fragmentos de ADN genómico de lulo fueron aislados por PCR, secuenciados y ensamblados en una secuencia parcial de 1417 pb (SquPPO1) que carece de intrones. La hibridación de una sonda de 920 pb generada a partir de un gen ppo de papa con una región de 12 kb de ADN de lulo digerido con BamHI-PstI, BamHI-XbaI y XbaI-PstI confirmó la presencia de al menos un gen ppo en esta especie. Mientras que se han identificado dos sitios conservados (Tyr-1 y Tyr-2) en los dominios de unión a cobre de otras PPOs de solanáceas, no se encontró ningún sitio Tyr-2 en la PPO de lulo debido a una sustitución conservativa DxE en esta región. El análisis filogenético situó al gen SquPPO1 en el mismo grupo que los genes SmePPO4, SmePPO5 y SmePPO6 de la berenjena (Solanum melongena L.). Nuestros resultados muestran que SquPPO1 es filogenéticamente más cercano a los genes ppo de berenjena que a los de papa, tabaco y tomate, y que presenta una variación que modifica la distribución de los sitios conservados de la proteína. Estos hallazgos ofrecen nuevos conocimientos sobre las bases moleculares del pardeamiento enzimático en lulo y pueden servir de base a estrategias para reducir las pérdidas poscosecha.
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