Overexpression of L-galactono-1,4-lactone dehydrogenase (L-GalLDH) gene correlates with increased ascorbate concentration and reduced browning in leaves of Lactuca sativa L. after cutting

Abstract

Ascorbic acid (AA) is one of the most powerful natural antioxidant able to prevent enzymatic browning after exogenous treatment of minimally-processed products. The specific mechanism by which AA prevents enzymatic browning remains still debated and a direct effect of endogenous AA stimulation and browning has never been studied. The manipulation of AA pathway is a promising approach to study the biochemical mechanism by which AA acts as an anti-browning agent. In this work, cDNA of L-galactono-1,4-lactone dehydrogenase (L-GalLDH), one of the key gene of the Smirnoff–Wheeler’s branch of AA biosynthetic pathway, was isolated and overexpressed in lettuce (Lactuca sativa L. cv ‘Iceberg’), a species highly prone to browning. The hypothesis that the overexpression of L-GalLDH translates to AA accumulation and reduces the browning phenomena in lettuce leaves after cutting was tested. Our results indicate that transgenic lettuce plants, showing about 19-fold overexpression of L-GalLDH as compared to wild type, had about +30 % of AA concentration in mature leaves. Transgenic plants exhibited reduced browning over the leaves, even 10 day after cutting, as demonstrated by higher values of luminosity (L*) and higher values of greenness (a*) compared to control plants. Overall, these findings provide a first evidence of the role of endogenous AA as browning-preventing agent. The obtainment of T2 transgenic lettuce plants is a promising first step to further determine the specific mechanism by which AA acts as an anti-browning preservative.