L-phenylalanine applications and culture duration affect root growth and production of tropane alkaloids and phenolics in adventitious root cultures of Hyoscyamus niger L.

Abstract

This study was aimed to determine the effects of different concentrations of L-phenylalanine (0.25, 0.50 and 1.00 mM L-Phe) applied to adventitious root cultures for different culture durations on root growth and production of tropane alkaloids and phenolics in Hyoscyamus niger. For this purpose, adventitious roots obtained from petiole explants of in vitro seedlings were used as plant materials. After adventitious roots were cultured in liquid MS medium, L-Phe was applied to the adventitious roots. Then the roots were harvested on the 1st, 3rd, and 7th day of the applications, fresh and dry root weights, root growth indexes were analysed. It was also examined the contents of tropane alkaloids (hyoscyamine and scopolamine) and phenolics (gallic acid, catechin, epicatechin, vanillin, cinnamic acid, rosmarinic acid, p-coumaric, ocoumaric acid, rutin, caffeic acid, ferulic acid, chlorogenic acid, and quercetin) by HPLC in both adventitious roots and culture medium. As a result, the greatest root growth values were obtained from the roots harvested on the 3rd and 7th days after 0.50 and 1.00 mM L-Phe applications. The maximum scopolamine and hyoscyamine amounts were detected in the cultures with the application of 0.50 mM L-Phe for 3 days. In terms of phenolics, 0.50 mM and 1.00 mM L-Phe application with 1st and 3rd days were selected as the most appropriate L-Phe applications and culture duration. Consequently, it has been determined that in vitro secondary metabolite production may be increased in H. niger root cultures with the selection of appropriate L-Phe concentrations and culture durations.

Effect of culture time on secondary metabolite production from MeJA-stimulated in vitro roots of Hyoscyamus niger L.

“…Seed germination and adventitious root collection followed the methods described in detail in our previous study (Demirci et al, 2022). Briefly, seeds treated with gibberellic acid for 48 hours were rinsed with double distilled water (ddH2O) and then sterilized by shaking with 70% ethanol (%99, Tekkim, Turkey) for 10 seconds and with 0.1% mercury chloride (HgCl2, Sigma-Aldrich, Germany) for 10 min.…”

“…Culture time is another important factor determining yield in the in vitro production of plant SMs (Demirci et al, 2020;Demirci et al, 2021;Demirci et al, 2022;Narayani and Srivastava 2017). In nature, each SM reaches its highest level at different time periods (Figueiredo et al, 2008;Soni et al, 2015).…”

Total Content of Saponins, Phenols and Flavonoids and Antioxidant and Antimicrobial Activity of In Vitro Culture of Allochrusa gypsophiloides (Regel) Schischk Compared to Wild Plants

“…A number of studies revealed that the duration of cultivation on the inducing medium also affects the content of secondary metabolites and their qualitative composition in the in vitro AR culture. The dependence of tropane alkaloid and phenol accumulation on the duration of cultivation and L-phenylalanine treatment in the in vitro AR culture of Hyoscyamus niger L. was shown [52]. The effect of the culture period on the yield of secondary metabolites with an optimal 15-day period is shown in the in vitro AR of Hyoscyamus niger L. [53].…”