Optimization of culture conditions and cultivation phase for the growth of Salvia viridis transformed roots and polyphenolic compound production

Izabela Grzegorczyk-Karolak

doi:10.1007/s11240-020-01883-6

Izabela Grzegorczyk-Karolak

Open Access

https://doi.org/10.1007/s11240-020-01883-6

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Abstract

Salvia viridis transformed root culture was studied using MS, WP or B5 media (full or half-strength macro- and microelements: 1/2MS, 1/2WP, 1/2B5) under varying light conditions to optimize growth and polyphenolic compound production. The highest levels of root growth (154.6 g/l and 13.6 g/l, respectively, fresh and dry weight) were observed for hairy roots grown in WP medium in the dark. UPLC analysis found the polyphenolic acid content in plant material to range from 20.7 to 45.9 mg/g dry weight (DW) depending on the conditions. The highest metabolite content was observed in roots grown in B5 medium under photoperiods. A time-course study of the hairy roots cultured in WP medium in the dark examined the effect of culture period on biomass and polyphenolic compound yield every 5 days over a 60-day period. More than 1 g of dry weight per flask and 47.8 mg of metabolite per g DW were produced after 30 days, at the beginning of the stationary phase. However, maximum production was only observed for four of the 10 detected compounds during this growth phase, between days 30 and 40; one of which was rosmarinic acid: the predominant compound in all extracts. Under optimal conditions, production was almost 10 times higher than that observed in the roots of field growing plants, which suggests that the selected optimized transformed root culture of S. viridis may be a good approach for valuable polyphenol production.

Highlights

Salvia viridis L. has long been used as a herb in Turkey and Iran, especially against inflammatory and infectious diseases (Mosaddegh et al 2012)
In vivo and in vitro experiments have demonstrated that salvianolic acids promote the apoptosis of cancer cells and inhibit the epithelial–mesenchymal transition processes associated with cancer
Six different media (WP, B5, MS, 1/2WP, 1/2B5, 1/2MS) were tested under dark and photoperiod conditions to optimize the growth of S. viridis transformed root culture

Summary

Introduction

Salvia viridis L. has long been used as a herb in Turkey and Iran, especially against inflammatory and infectious diseases (Mosaddegh et al 2012). Several classes of secondary metabolites have been isolated from the aboveground and underground parts of the plant, including polyphenolic acids, phenylethanoids, flavonoids (Rungsimakan and Rowan 2014; Grzegorczyk-Karolak and Kiss 2018), triterpenes Plant Cell, Tissue and Organ Culture (PCTOC) (2020) 142:571–581. Hairy root culture of S. viridis found to be a potentially rich source of valuable polyphenolic acids, such RA and salvianolic acids (Grzegorczyk-Karolak et al 2018). In a selected clone (K3), RA level was found to be 8-fold higher than in the roots of field-grown plants. This culture was characterized by high levels of salvianolic acids E and F, which are found only in small quantities in the roots of field-grown S. viridis. In vivo and in vitro experiments have demonstrated that salvianolic acids promote the apoptosis of cancer cells and inhibit the epithelial–mesenchymal transition processes associated with cancer

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