Elicitation of Submerged Adventitious Root Cultures of Stevia rebaudiana with Cuscuta reflexa for Production of Biomass and Secondary Metabolites

Abstract

Stevia rebaudiana is an important medicinal plant that belongs to the Asteraceae family. The leaves of Stevia rebaudiana are a rich source of many health-promoting agents such as polyphenols, flavonoids, and steviol glycoside, which play a key role in controlling obesity and diabetes. New strategies such as the elicitation of culture media are needed to enhance the productivity of active components. Herein, the Cuscuta reflexa extracts were exploited as elicitors to enhance the productivity of active components. Cuscuta reflexa is one of the parasitic plants that has the ability to elongate very fast and cover the host plant. Consequently, it may be possible that the addition of Cuscuta reflexa extracts to adventitious root cultures (ADR) of Stevia rebaudiana may elongate the root more than control cultures to produce higher quantities of the desired secondary metabolites. Therefore, the main objective of the current study was to investigate the effect of Cuscuta reflexa extract as a biotic elicitor on the biomass accumulation and production of antioxidant secondary metabolite in submerged adventitious root cultures of Stevia rebaudiana. Ten different concentrations of Cuscuta reflexa were added to liquid media containing 0.5 mg/L naphthalene acetic acid (NAA). The growth kinetics of adventitious roots was investigated for a period of 49 days with an interval of 7 days. The maximum biomass accumulation (7.83 g/3 flasks) was observed on medium containing 10 mg/L extract of Cuscuta reflexa on day 49. As the concentration of extract increases in the culture media, the biomass gradually decreases after 49 days of inoculation. In this study, the higher total phenolics content (0.31 mg GAE/g-DW), total flavonoids content (0.22 mg QE/g-DW), and antioxidant activity (85.54%) were observed in 100 mg/L treated cultures. The higher concentration (100 mg/L) of Cuscuta reflexa extract considerably increased the total phenolics content (TPC), total phenolics production (TPP), total flavonoids content (TFC), total flavonoids production (TFP), total polyphenolics content (TPPC), and total polyphenolics production (TPPP). It was concluded that the extract of Cuscuta reflexa moderately improved biomass accumulation but enhanced the synthesis of phenolics, flavonoids, and antioxidant activities. Here, biomass’s independent production of secondary metabolites was observed with the addition of extract. The present study will be helpful to scale up adventitious roots culture into a bioreactor for the production of secondary metabolites rather than biomass accumulation in medicinally important Stevia rebaudiana.

Biological synthesis of hybrid silver nanoparticles by Periploca aphylla Dcne. From nanotechnology to biotechnology applications

“…Many studies on the in vitro propagation, multiplication, and callogenesis of S. rebaudiana have been undertaken in the past, but there is little information available on callus proliferation and the induction of antioxidants and bioactive chemicals that are helpful to human health ( Ahmad et al, 2013 ; Ahmad et al, 2022 ).…”

Time and Concentration-Dependent Differential Antioxidant Potential in the Gum of Medicinally Important Araucaria Heterophylla

“…Normally the combined solution of any plant extract and DPPH free radicals' solution was incubated in a dark room to avoid oxidation and to form stable complexes of free radicals with plant-based materials (Ahmad et al 2012;Ali et al 2016;Khalil et al 2016;Fazal et al 2016b;Ahmad et al 2022). Most of the literature suggested 30 min incubation in the dark and further increments in the incubation period display a negative impact on antioxidant activity (Ali et al 2017;Ghazal et al 2018;Nawaz et al 2018;Idrees et al 2018).…”

“…In contrast, the gum of Araucaria heterophylla was found to have good cytotoxic activity against HGE cells (human gastric epithelial) (Elkady and Ayoub 2018;Frezza et al 2020). Due to oxidation damage that leads to many diseases such as Parkinson's disease, inflammation, aging, liver diseases, arthritis, Alzheimer's disease, and cancer (Fazal et al 2020;Ahmad et al 2020;Ahmad et al 2022;Ilyas et al 2022). Antioxidant components can inhibit oxidative damage caused by free radicals (Ahmad et al 2021).…”

Melatonin-Induced Stress Enhanced Biomass and Production of High-Value Secondary Cell Products in Submerged Adventitious Root Cultures of Stevia rebaudiana (Bert.)

“…Elicitation of culture media with different types of stressors or elicitors (biotic or abiotic) may enhance the productivity of metabolites of interest. , Elicitation strategies are gaining more industrial interest for uniform and higher productivity of commercial secondary cell products . Elicitation strategies are commonly employed for high-value medicinal plants.…”