Elicitation of Stilbenes and Benzofuran Derivatives in Hairy Root Cultures of White Mulberry ( Morus alba ).

Stilbene and benzofuran derivatives isolated from the root of white mulberry ( Morus alba ) have shown various biological activities, including anti-inflammatory, antioxidant, and antimicrobial properties. The objectives of this study were to develop hairy root cultures and assess the effect of multiple elicitors combinations including (I) methyl-β-cyclodextrin (CD), MgCl 2 , methyl jasmonate (MeJA), and H 2 O 2 , (II) CD, MgCl 2 , and MeJA and (III) CD, MgCl 2, and H 2 O 2 , on the production of these bioactive compounds. The highest yields of stilbenes and benzofurans were obtained upon co-treatment with 18 g/L CD, 3 mM H 2 O 2 and 1 mM MgCl 2 . The stilbenes oxyresveratrol, resveratrol, and 3'-prenylresveratrol accumulated up to 6.27, 0.61, and 5.00 mg/g DW root, respectively. Meanwhile, the aryl benzofurans moracin M and moracin C accumulated up to 7.82 and 1.82 mg/g DW root, respectively. These stilbenes and benzofurans accumulated in the culture medium of the elicited hairy root cultures. They were not detected in the root tissue. However, the oxyresveratrol diglucoside mulberroside A was only detected in the root tissue with yields up to 10.01 mg/g DW. The results demonstrated that co-treatment of white mulberry hairy root cultures with multiple elicitors can significantly enhance production and secretion of stilbenes and benzofurans in this controlled and sustainable axenic culture system.