Differential Production of Tropane Alkaloids in Hairy Roots and in vitro Cultured Two Accessions of Atropa belladonna L. under Nitrate Treatments

Abstract

Plants are a potential source of a large number of valuable secondary metabolites. In vitro cultures are being considered as an alternative to agricultural processes for studying valuable secondary metabolites. In this way, nutritive factors are important parameters influencing the production of these compounds in plants. Effects of nitrate concentrations (KNO3) on the production of two tropane alkaloids, hyoscyamine and scopolamine, and the growth of aerial parts and roots of two in vitro propagated accessions of Atropa belladonna and hairy roots were investigated. As hairy roots cultures are able to keep a stable production of alkaloids over long periods of subculturing, they are considered as an interesting option for the study of alkaloid biosynthesis. A hairy roots culture of Atropa belladonna was established by transformation with Agrobacterium rhizogenes strain AR15834. The results of our study showed that a rise in KNO3 concentration caused a decline in hairy roots growth, and had a remarkable effect on the alkaloid content. The alkaloid concentrations obtained in the hairy roots were 3 - 20 times higher than that in the plants at 35 mM of KNO3. Increasing the nitrate concentration in the medium of hairy roots also improved the hyoscyamine/scopolamine ratio, while it increased the scopolamine/hyoscyamine ratio in the studied plants.

Rapid, sensitive, and validated UPLC/Q-TOF-MS method for quantitative determination of vasicine in Adhatoda vasica and its in vitro culture

“…The results obtained with NaCl treatment [Figure 6c] were found in accordance to the earlier reports. [171819212223]…”

“…Regular subculturings were done at every 3 weeks for proper growth of calli. The initiated calli on MS medium supplemented with growth hormones showing best results were chopped and transferred aseptically into flasks containing MS medium prepared afresh with same hormonal combination and varying concentration of potasium nitrate (20-35 mM)[17] and NaCl (25-200 mM)[1819] to check their effect on biomass and vasicine production. All the cultures were maintained for 120 days with regular growth monitoring and subculturing after every 3 weeks, which were harvested and analyzed for vasicine contents.…”

Increasing the Pace of New Discoveries in Tropane Alkaloid Biosynthesis

“…Salicylic acid (SA), another common plant defense hormone, is often antagonistic to MeJA-induced responses (Niki, Mitsuhara, Seo, Ohtsubo, & Ohashi, 1998). For example, increasing nitrate concentrations in A. belladonna hairy root cultures yielded higher amounts of TAs, at the same time reducing the overall growth of the culture (Chashmi, Sharifi, Karimi, & Rahnama, 2010). The use of heavy metal elicitors has also been shown to successfully increase the production of TAs.…”

In vitro production of alkaloids: Factors, approaches, challenges and prospects

“…[17] The rise in KNO 3 concentration up to 35 mM increased the tropane alkaloidal content up to 3-20 times with improved ratio of hyoscyamine/scopolamine. [18] In vitro grown tissues (non-regenerative callus, regenerative callus, and microshoot-derived leaves) of Solanum nigrum cultured under salinity stress (0-150 mM) were found to yield maximum of solasodine with 150 mM of sodium chloride. [19]…”