Improved production of dibenzocyclooctadiene lignans in the elicited microshoot cultures of Schisandra chinensis (Chinese magnolia vine)
Improved production of dibenzocyclooctadiene lignans in the elicited microshoot cultures of Schisandra chinensis (Chinese magnolia vine)
Agnieszka Szopa 0 1
Halina Ekiert 0 1
Adam Kokotkiewicz 0 1
Agata Król 0 1
Maria Luczkiewicz 0 1
0 Department of Pharmacognosy, Faculty of Pharmacy, Medical University of Gdańsk , al. gen. J. Hallera 107, 80-416 Gdańsk , Poland
1 Department of Pharmaceutical Botany, Jagiellonian University , Collegium Medicum, ul. Medyczna 9, 30-688 Kraków , Poland
Dibenzocyclooctadiene lignans are a specific group of secondary metabolites that occur solely in Schisandra chinensis. The aim of the presented work was to boost the accumulation of lignans in the agitated microshoot cultures of S. chinensis, using different elicitation schemes. The experiments included testing of various concentrations and supplementation times of cadmium chloride (CdCl2), chitosan (Ch), yeast extract (YeE), methyl jasmonate (MeJa), and permeabilizing agent-dimethylsulfoxide (DMSO). After 30 days, the microshoots were harvested and evaluated for growth parameters and lignan content by LC-DAD method. The analyses showed enhanced production of lignans in the elicited S. chinensis microshoots, whereas the respective media samples contained only trace amounts of the examined compounds (< 5 mg/l). Elicitation with CdCl2 caused up to 2-fold increase in the total lignan content (max. ca. 730 mg/100 g DW after the addition of 1000 μM CdCl2 on the tenth day). Experiments with chitosan resulted in up to 1.35-fold increase in lignan concentration (max. ca. 500 mg/100 g DW) after the supplementation with 50 mg/l on the first day and 200 mg/l on the tenth day. High improvement of lignan production was also recorded after YeE elicitation. After the elicitation with 5000 mg/l of YeE on the first day of the growth period, and with 1000 and 3000 mg/l on the 20th day, the lignan production increased to the same degree-about 1.8-fold. The supplementation with 1000 mg/l YeE on the 20th day of the growth cycle was chosen as the optimal elicitation scheme, for the microshoot cultures maintained in Plantform temporary immersion system-the total content of the estimated lignans was equal to 831.6 mg/100 g DW.
Elicitation; Temporary-immersion bioreactor; Schisandra lignan production; Biotic elicitors; Abiotic elicitors; In vitro cultures
Introduction
Schisandra chinensis (Turcz.) Baill., Chinese magnolia vine
(Schisandraceae), is a climbing plant, naturally occurring in the
countries of Eastern Asia, whereas in European countries, it
grows mainly as an ornamental shrub
(Panossian and Wikman
2008; Szopa et al. 2016a; Szopa et al. 2017a)
. The raw material
of Schisandra fruits, Schisandrae chinensis fructus (chin. běi wǔ
wèi zi; literally Bfive-flavor berry^), has been used for therapeutic
purposes in traditional Chinese medicine and has successfully
been included in pharmacopoeial monographs of Asian and
European countries as well as in the USA Pharmacopoeia and
the International Pharmacopoeia, printed by WHO
(World
Health Organization 2007; European Directorate for the Quality
of Medicines. 2017; Szopa et al. 2017a)
. Numerous therapeutic
properties of Schisandra fruit extracts, as well as its individual
compounds, have been confirmed by the scientific research,
carried out with the use of in vivo and in vitro models. The studies
indicate that S. chinensis has a positive effect on liver functioning
and stimulates cardiovascular, respiratory, and central nervous
systems. The therapeutic properties of Chinese magnolia vine
include anticancer, immunostimulant, and adaptogenic
(Mocan
et al. 2016; Szopa et al. 2016a; Szopa et al. 2017a)
. These
activities are related to the presence of the dibenzocyclooctadiene
lignans, known as BSchisandra lignans^ (Figure S1), due to the
fact that their occurrence is limited to S. chinensis
(Fuss 2004;
Opletal et al. 2004; Lu and Chen 2009)
.
The dominant lignans that can be distinguished out of over
40 present in S. chinensis are as follows: schisandrin; gomisins
A, C, F and N; deoxyschisandrin; and γ-schisandrin. The
extracts of S. chinensis fruit have been used for the manufacture
of medical products, food supplements, or cosmetics
(Szopa
et al. 2016a)
. Given the fact that the lignans of S. chinensis are
therapeutically unique compounds, the research on alternative
methods of their natural resource-independent acquisition has
been performed. The methods of the chemical synthesis of
Schisandra lignans have been developed; however, these
had limited success due to their complex stereochemistry
(Shi et al. 2009)
. The other alternative is plant cell culture
which has so far been employed for the production of several
biologically active secondary metabolites, including lignans.
Moreover, it has been reported that in vitro techniques enable
to enhance the production of lignans by means of elicitation,
addition of biosynthetic precursors, and immobilization
(Angelova et al. 2006; Ver (...truncated)