Constitutive production of trans-resveratrol via callus induction from Vitis vinifera cells

CHIALVA CS; ORDEN AA; PUNGITORE CR; TONN CE

Mendoza

Congreso; Sociedad Biología de Cuyo; 2010

Sociedad de Biología de Cuyo

Grapevine (Vitis spp.) is one of the most important fruit crops cultivated worldwide. Its more relevant phytoalexins comprise a group of molecules belonging to the stilbene family such as trans-resveratrol (3,5,4´-trihydroxystilbene). Consequently, we undertook the production of trans-resveratrol using Vitis vinifera L. cell cultures in order to investigate its constitutive production. Explants from Red Globe vineyard plants were excised from leaves and stems prior to surface sterilization. The cuttings were placed on autoclaved solid MS medium containing growing regulators such as 0,5 ppm ANA; 1:0,1 ppm BAP/2,4-D; 4:4 ppm IBA/BAP; 2:0,1 ppm ANA/Kin; 1 ppm 2,4-D; and 2-3% sucrose concentrations. Cultures were incubated at 22 ±2°C in the dark and were transferred every 4 weeks to fresh medium. Liquid medium was used for the production of secondary metabolites. The extracts for analysis were obtained from the growing medium by liquid-liquid partition with EtOAc. Samples were analized by GC prior to permethylation using (CH3)2SO4. The composition of the MS medium supplemented with 1:0,1 ppm BAP/2,4-D and 2% of sucrose was found suitable for this aim. Indeed, grape cell suspension cultures synthesized detectable levels of trans-resveratrol. Results show an appropiate method for callus induction and culture maintenance with production of the stilbene resveratrol. We will searching for some elicitors that could yield a higher resveratrol accumulation than without induction.