Impact of light and sucrose supplementation on cellular differentiation, metabolic shift and modulation of gene expression in hairy roots of Daucus carota

Abstract

Green hairy root cultures of Daucus carota were subjected to gradually lowered sucrose concentrations to understand the modulation of secondary metabolism upon carbon-starved conditions. An apparent negative correlation with the sucrose concentrations in the media was observed with chloroplast development, as confirmed by both histological studies and relevant biochemical assays. Expansion of relative vascular area and well-developed multi-layered endodermis were evidenced in green hairy roots with low sucrose concentrations as compared to normal hairy roots cultivated with 2% sucrose under dark condition. Thus, anatomical modulation was evident in photo-oxidatively stressed green hairy roots because of their maintenance under continuous illumination. This was demonstrated further by the expression analysis of glutathione peroxidase and NADPH reductase genes which showed upregulation in green hairy roots maintained in ½ MS media containing 0.5% sucrose as compared to normal hairy roots. The amount of phenolic marker compound p-hydroxybenzoic acid was reduced to half, while enhanced levels of emitted terpenoid volatiles were noticed in these carbon-starved green hairy roots. Analysis of primary metabolites in green hairy roots showed increased levels of amino acids and sugar alcohols accumulation. Further, analysis revealed the elevated accumulation of organic acids which have significant bearing towards redirection of primary metabolites leading to the formation of enhanced terpenoid volatiles in green hairy roots. Thus, our study revealed the impact of light and sucrose supplementation towards cellular differentiation and redirection of secondary metabolism in hairy roots of D. carota.