Secondary Metabolites of Medicinal Plants. Bharat Singh

and/or nicotine in A. belladonna (Endo and Yamada 1985; Yang et al. 2011). By HPLC determination, it was concluded that the accumulation of tropane alkaloids was higher in in vitro regenerated leaves but lower in the roots and shoots. Similarly, the atropine accumulation was higher, but scopolamine accumulated in lower concentration in the leaves of A. acuminata (Banerjee et al. 2008; Ashtiania and Sefidkonb 2011). The transgenic cells (containing H6H gene/s) of A. baetica were elicited with salicylic acid, acetyl salicylic acid, and methyl jasmonate for the induction of synthesis of tropane alkaloids. The synthesis of scopolamine was increased by the acetyl salicylic acid and methyl jasmonate, while salicylic acid did not affect the synthesis. This happened due to the expression of engineered h6h genes and other genes involved in the biosynthesis of alkaloids. The synthesis of scopolamine was increased by 25-fold higher than in control (Jaber-Vazdekis et al. 2008).

      The biotransformation of betuligenol into an oxidized product betuloside was performed in the hairy roots of A. belladonna. The yield of biotransformed products increased twofold higher than the control root mass. On the 5th and 10th days of incubation, the highest biotransformation of betuligenol into raspberry ketone and betuligenol into betuloside was reported in hairy roots. The application of hairy roots for biotransformation of raspberry ketone and betuloside opens up new opportunities in the production of medicinally significant secondary metabolites (Srivastava et al. 2013).

The pharmaceutically important tropane alkaloids were isolated from the A. belladonna. The putrescine N-methyltransferase from N. tabacum and hyoscyamine 6β-hydroxylase from Hyoscyamus niger were overexpressed in transgenic A. belladonna for investigation of biosynthesis of tropane alkaloids in stem roots and fruits. The expression of putrescine N-methyltransferase and hyoscyamine 6β-hydroxylase was not reported in wild plant species but were found transgenic plants. The maximum conversion of hyoscyamine was observed in aerial parts, not in underground parts. In aerial parts, the hyoscyamine was fully transformed into scopolamine. The synthesis of hyoscyamine, anisodamine, and scopolamine was reported to have higher yield in underground parts than in aerial parts (Xia et al. 2016).

      The effects of KCr(SO₄)₂ on the accumulation of tropane alkaloids as well as the expression of hyoscyamine 6β-hydroxylase gene was investigated in in vitro regenerated plantlets of A. belladonna. The chromium treatment to plantlets decreased the weights, lengths of the plantlets, and chlorophyll contents but enhanced the levels of proline contents. The chromium treatment also increased the concentration of hyoscyamine and scopolamine. The levels of scopolamine can be correlated with the expression levels of h6h gene with several concentrations of chromium (Jaber-Vazdekis et al. 2009; Vakili et al. 2012).

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