Cyclodextrins and coronatine (Cor) are new elicitors and permeabilizing agents which are applied in plant cell and hairy root cultures to increase phytopharmaceoticals production (1). In present research the effect of two biotic elicitors, methyl-β- cyclodextrin (β-CD) and Cor, on hairy root cultures of A. acuminata Royle ex Miers was evaluated in order to develop a sustainable and high production system of hyoscyamine (HYO) and especially scopolamine (SCO) tropane alkaloids. In this regard, the Agrobacterium rhizogenes A4 was used for direct infection of 2 month pot plant leaves of A. acuminata. Inoculums as 1 gr fresh weight of selected hairy root line were inoculated in glass jars which contained 30 ml of liquid half-strength B5 (B5/2) medium supplemented with 3% sucrose and placed in a shaker incubator (110 rpm) at 25 C° in dark situation. On day 14th the old medium was replaced with 30 ml fresh liquid medium as the same explained above which containing the elicitor treatments including: 50 mM β-CD, 0.5 μM Cor and joint treatment of 50 mM β-CD and 0.5 μM Cor with 3 replications. The sampling was fulfilled on day 21 (T1) and 28 (T2) of culture. HPLC analysis showed that Cor had negative effect on HYO content and also β-CD alone or in combination with Cor didn’t have any significant effect on HYO content in hairy root cultures. On the other hand, the maximum amount of SCO was achieved by Cor alone at T1 and it was about 10 times more than controls at both sampling times. Also joint treatment of β-CD and Cor at T1 and T2 had positive effect on SCO quantity (Fig. 1).
Key words: Cyclodextrin, Coronatine, Hairy root, Atropa acuminata
Reference
1- Khojasteh A, Mirjalili, M. H, Hidalgo D, Corchete P, Palazon J. Biotechnol Lett 2014; 36: 2393-2406