The effect of light, phenylalanine and methyl jasmonate, alone or in combination, on growth and secondary metabolism in cell suspension cultures of Vitis vinifera

V. vinifera is a rich source of stilbene compounds such as resveratrol, which are widely believed to provide dietary protection against the development of cardiovascular disease and some forms of cancer [1]. Elicitation is a well-known strategies to increase commercial production of natural products in plant cell suspension culture systems [2]. Hence, in current study, we investigated the effect of two light regimes (10,000 LUX radiation and dark), amino acid aromatic phenylalanine (Phe) precursor (0, 0.1, 0.5 and 1 mM) and methyl jasmonate elicitor (0 and 25 μM), alone or in combination on stilbene biosynthesis in cell suspension cultures of V. vinifera. Callus tissues obtained from the berries of V. vinifera cv. Shahani on an optimized medium (MS microelements, B5 macroelements and Morel vitamins) containing 20 g/L sucrose, 250 mg/L casein hydrolysate, 0.2 mg/L kinetin and 0.5 mg/L α-naphthalene acetic acid were used to develop cell suspension cultures. The results showed that cultures grown in darkness resulted in significantly higher level of total stilbenes (resveratrol + piceid) (58.11 μg/g DW) accumulation when compared with high-level light condition (21.03 μg/g DW). The combined treatments of dark + 1 mM Phe and dark + 25 μM MeJA highly induced total phenolics, total flavonoids and total stilbenes bio-production to the maximum of 241.16-279.47 mg GA/g DW, 310.46-378.14 mg catechin/g DW, and 98.54-110.54 μg/g DW, respectively. Finally, the combined elicitation of dark + 1 mM Phe + 25 μM MeJA with the highest synergistic coefficient (1.24) proved to be the most effective for production of total phenolics, total flavonoids, and total stilbenes with mean contents of 384.80 mg GA/g DW, 527.62 mg catechin/g DW and 188.34 μg/g DW, respectively. Additionally, there was a significant negative correlation between cell growth and the production of phenolic compounds in cells subjected to different treatments. The results of our study suggest that the combinations of dark together with MeJA and/or Phe can be used as an efficient method for the future scale-up of V. vinifera cell cultures for the production of high value stilbene compounds in a bioreactor system.

Keywords:

Vitis vinifera , Elicitation , Dark , Phenylalanine , Methyl jasmonate , Stilbene compounds