Phytochemical and biotechnological studies on Schisandra chinensis cultivar Sadova No. 1-a high utility medicinal plant

Abstract

In the presented work, raw materials (fruits and leaves) and in vitro biomass of a highly productive Schisandra chinensis Sadova No. 1 cultivar (SchS) were evaluated for the production of therapeutically useful schisandra lignans (SL). In vitro cultures of SchS were initiated, followed by extensive optimization studies focused on maximizing secondary metabolite production, with the aim of establishing a sustainable source of SL. Different cultivation systems (agar, agitated, bioreactor), experiment times (10, 20, 30, 40, 50 and 60 days) and plant growth regulators (6-benzyladenine-BA and 1-naphthaleneacetic acid-NAA, from 0 to 3 mg/l) in Murashige-Skoog (MS) medium were tested. Moreover, an elicitation procedure was applied to bioreactor-grown microshoots in order to increase SL production. Validated HPLC-DAD protocol enabled to detect fourteen SL in the extracts from in vitro and in vivo materials. The main compounds in the in vitro cultures were as follows: schisandrin (max. 176.3 mg/100 g DW), angeloylgomisin Q (max. 85.1 mg/100 g DW), gomisin A (max. 71.4 mg/100 g DW) and angeloylgomisin H (max. 67.0 mg/100 g DW). The highest total SL content (490.3 mg/100 g DW) was obtained in extracts from the biomass of agar cultures cultivated for 30 days on the MS medium variant containing 3 mg/l BA and 1 mg/l NAA. This amount was 1.32 times lower than in fruit extracts (646.0 mg/100 g DW) and 2.04 times higher than in leaf extracts (240.7 mg/100 g DW). The study demonstrated that SchS is a rich source of SL, thus proving its value for medical, cosmetic and food industry.

Keywords: Agar culture; Agitated culture; Bioreactor culture; Chinese magnolia vine cultivar; Elicitation; In vitro cultures; Schisandra lignans.

Fig. 1

Morphological appearance of S. chinensis cv. Sadova microshoots grown for 10–60 days on different MS (Murashige and Skoog) agar media variants: A—control (without PGRs), B—0.1 mg/l BA and 2 mg/l NAA, C—0.5 mg/l BA and 2 mg/l NAA, D—2 mg/l BA and 0.5 mg/l NAA, E—2 mg/l BA and 1 mg/l NAA, F—2 mg/l BA and 2 mg/l NAA, G—3 mg/l BA and 1 mg/l NAA

Fig. 1

Morphological appearance of S. chinensis cv. Sadova microshoots grown for 10–60 days on different MS (Murashige and Skoog) agar media variants: A—control (without PGRs), B—0.1 mg/l BA and 2 mg/l NAA, C—0.5 mg/l BA and 2 mg/l NAA, D—2 mg/l BA and 0.5 mg/l NAA, E—2 mg/l BA and 1 mg/l NAA, F—2 mg/l BA and 2 mg/l NAA, G—3 mg/l BA and 1 mg/l NAA

Fig. 2

Morphological appearance of S. chinensis cv. Sadova agitated microshoots grown for 10–60 days in different liquid media variants: F—2 mg/l BA and 2 mg/l NAA, G—3 mg/l BA and 1 mg/l NAA

Fig. 3

S. chinensis cv. Sadova microshoots grown for 30 days in temporary immersion system (Plantform bioreactor) on MS medium variant G (3 mg/l BA and 1 mg/l NAA)

Fig. 4

The representative HPLC-UV chromatogram (λ = 225 nm) of a methanol extract of S. chinensis cv. Sadova biomass cultivated in vitro. SL: 1—schisandrin, 2—gomisin, 3—angeloyl-/tigloylgomisin, 4—angeloyl-/tigloylgomisin Q, 5—gomisin G, 6—schisantherin A, 7—schisantherin B, 8—schisanthenol, 9—deoxyschisandrin, 10—schisandrin B, 11—γ-schisandrin, 12—benzoylgomisin P, 13—schisandrin, 14—schisantherin D

Fig. 5

The dynamic of schisandra lignans accumulation in agar S. chinensis cv. Sadova microshoots cultures based on their total contents with relation to the duration of growth period and applied MS medium variants

Fig. 6

The dynamic of schisandra lignans accumulation in agitated S. chinensis cv. Sadova microshoots cultures based on their total contents with relation to the duration of growth period and applied MS medium variants