Anethum graveolens L. In Vitro Cultures – a Potential Source of Bioactive Metabolites, Phenolic Acids and Furanocoumarins

Abstract

In vitro cultures of Anethum graveolens (dill) were maintained on the Linsmaier and Skoog (LS) medium – three variants, and the Murashige and Skoog (MS) medium – seven variants, which contained different amounts of plant growth regulators, cytokinin (BAP) and auxin (NAA) (from 0.1 mg l-1 to 3.0 mg l-1). Methanolic extracts from in vitro grown biomass were analyzed by HPLC for free phenolic acids and furanocoumarins. The total amounts of free phenolic acids on the LS medium variants were similar (35.23–38.65 mg 100 g-1 DW), but higher on the MS variants, ranging from about 66 mg 100 g-1 DW to 100 mg 100 g-1 DW. The main metabolites were: p-hydroxybenzoic acid (max. 24.41 mg 100 g-1 DW) on the LS-based media, and salicylic acid (max. 57.88 mg 100 g-1 DW) and p-hydroxybenzoic acid (max. 36.27 mg 100 g-1 DW) on the MS-based media. The total amounts of furanocoumarins were lower, as they did not exceed 8.5 mg 100 g-1 DW on the LS media and 25 mg 100 g-1 DW on the MS media. The main compounds in this group were bergapten (max. 15.01 mg 100 g-1 DW) and marmesin (max. 8.12 mg 100 g-1 DW). The MS variant containing 0.5 mg l-1 BAP and 2.0 mg l-1 NAA was proposed as the best production medium for both groups of metabolites. The maximum total amounts of free phenolic acids obtained in the in vitro grown biomass were slightly higher than their amounts in the fruits of the mother plant analyzed for comparison (99.66 mg 100 g-1 DW and 93.34 mg 100 g-1 DW, respectively); the maximum total amounts of furanocoumarins were approximately 1.8 times higher than in the fruits (24.26 mg 100 g-1 DW and 13.67 mg 100 g-1 DW, respectively).