In vitro cultivation of tansy (Tanacetum vulgare L.): a tool for the production of potent pharmaceutical agents

Abstract

In this study, tansy (Tanacetum vulgare L.) in vitro culture was established from seeds collected from natural populations. The multiplication of plantlets was conducted through shoot tips that exhibited potent apical growth and regeneration capacities on basal medium (BM), without the addition of any plant growth regulators (PGRs). PGRs were also omitted for the establishment and cultivation of tansy root cultures. Both abaxial and adaxial leaf surfaces of in vitro micropropagated plantlets were covered with glandular biseriate trichomes. Histochemical staining showed that glandular secretions were rich in lipid and terpene compounds, confirmed by GC-MS analysis of essential oil (EO). In the total EO, similar portions of oxygenated monoterpenes (38.5% m/m) and oxygenated sesquiterpenes (22.6% m/m) were detected. Chemical profiles of methanol extracts of in vitro cultured tansy shoots and roots varied in quantity and quality from those obtained from wild-growingtansy. HPLC analysis indicated that the methanol extracts of in vitro cultured roots were the richest in 3,5-O-dicaffeoylquinic acid (3,5-O-DCQA), in which the concentration was 6 times higher (10.220 mg/g DW) than that in the extract obtained from roots of wild-growing tansy (1.684 mg/g DW). This result is noticeable in the manner of industrial production of biologically active 3,5-O-DCQA that has been shown to have antioxidant, hepatoprotective, antiviral, antimutagenic, and immunomodulatory activity. Biotechnological interventions on secondary metabolite production taking place in trichomes could further enhance the production of some important tansy metabolites and further investigation will be directed toward the elucidation of the pharmaceutical potential of tansy in vitro obtained metabolites, as mixtures or single moieties.

Keywords: Essential oil; Histochemical analysis; In vitro cultivation; Methanol extracts; Phytochemical analysis; Tansy.

Fig. 1

In vitro micropropagation of tansy. a Seedlings observed with binocular magnifier; bar = 1 mm; b Petri dish with germinated seedlings; bar = 2 cm; c Nicely develop tansy seedling with 4–5 leaves and branched root; bar = 2 cm; d Tansy shoots grown on solid medium; e Erlenmeyer with tansy roots grown in liquid medium

Fig. 2

Scanning electron micrographs of in vitro grown tansy foliar surface. a Adaxial leaf surface; b abaxial leaf surface; c glandular (arrow) and non-glandular trichomes on the adaxial leaf surface; d glandular (arrow) and non-glandular trichomes on the abaxial leaf surface; e biseriate glandular trichome at the beginning of the secretory phase; f mature biseriate glandular trichome on the adaxial leaf surface in the full secretory phase

Fig. 3

Structural and histochemical features of leaf glandular trichomes from in vitro grown tansy. a Cross-section of tansy leaf, note: trichomes (arrow) on adaxial and abaxial leaf surface; b young, immature leaf glandular trichomes, note: two basal cells, a short stalk, and secretory head of three pairs cells; c unstained biseriate trichome with subcuticular space (arrow); d orange-brown colored of secretory material in subcuticular space after stained with Sudan Red 7B/hematoxylin; e dark-blue color of lipophilic substance after staining with Sudan black B; f neutral lipids/essential oils stained red, while acid lipids stained blue with Nil blue A; g–h positive reaction with NADI reagent, violet-blue droplets indicate terpene secretion; i positive reaction with PAS; j UV-autofluorescence micrographs of leaf glandular trichomes. Bar = 10 μm

Fig. 4

Chromatograms of tansy methanol extracts. Methanol extract of a native tansy herb; b in vitro grown tansy herb; c native tansy roots; d in vitro grown tansy roots. The arrows point to chlorogenic acid (CGA) and 3,5-O-dicaffeoylquinic acid (3,5-O-DCQA) peaks with given amounts of targeting compounds as mg per g of dry weight (DW) for each type of extracts