Genetic Stability, Phenolic, Flavonoid, Ferulic Acid Contents, and Antioxidant Activity of Micropropagated Lycium schweinfurthii Plants

Abstract

Lycium schweinfurthii is a Mediterranean wild shrub rich in plant secondary metabolites. In vitro propagation of this plant may support the production of valuable dietary supplements for humanity, introduction of it to the world market, and opportunities for further studies. The presented study aimed to introduce an efficient and reproducible protocol for in vitro micropropagation of L. schweinfurthii and assess the genetic stability of micropropagated plants (MiPs) as well as to estimate phenolic, flavonoid, ferulic acid contents, and the antioxidant activity in leaves of micropropagated plants. Two DNA-based techniques, random amplified polymorphic DNA (RAPD) and inter-simple sequence repeats (ISSR), and one biochemical technique, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), were used to assess the genetic stability in MiPs. Spectrophotometric analysis was performed to estimate total phenolic and flavonoid contents and antioxidant activity of MiPs leaves, while ferulic acid content was estimated using high-performance thin-layer chromatography (HPTLC). Sufficient shoot proliferation was achieved at MS (Murashige and Skoog) medium supplemented with 0.4 mg L−1 kinetin and rooted successfully on half-strength MS medium fortified with 0.4 mg L−1 Indole-3-butyric acid (IBA). The Jaccard’s similarity coefficients detected in MiPs reached 52%, 55%, and 82% in the RAPD, ISSR, and SDS-PAGE analyses, respectively. In the dried leaves of MiPs, the phenolic, flavonoid, and ferulic acid contents of 11.53 mg gallic acid equivalent, 12.99 mg catechin equivalent, and 45.52 mg were estimated per gram, respectively. However, an IC50 of 0.43, and 1.99 mg mL−1 of MiP dried leaves’ methanolic extract was required to scavenge half of the DPPH, and ABTS free radicals, respectively. The study presented a successful protocol for in vitro propagation of a valued promising plant source of phenolic compounds.

Callus-Mediated High-Frequency Plant Regeneration, Phytochemical Profiling, Antioxidant Activity and Genetic Stability in Ruta chalepensis L.

“…This may be explained by the fact that in vitro culture systems and PGRs enhance or accumulate several of the bioactive compounds. In vitro culture has been shown to increase the amounts of phytochemical compounds in many medicinal plants [15][16][17][42][43][44], which is consistent with our findings. In the Aloe arborescens, media supplemented with PGRs increased the quantity of total phenolics, tannins, and flavonoids compared to media without PGRs during micropropagation [45].…”

Micropropagation of Vaccinium corymbosum L.: An Alternative Procedure for the Production of Secondary Metabolites

“…Different groups of metabolites have been examined in the biomass of shoots, roots or calluses grown in vitro in several plant species such as: shoot cultures of Ruta graveolens [22], a biomass of shoots and differentiating callus culture of Schisandra chinensis [23], the shoots and roots of Eryngium planum [24], biomass from Aronia melanocarpa shoots and callus cultures [25], the leaves and roots of Rehmannia glutinosa [26], leaves of micropropagated plants of Lycium schweinfurthii [27], leaves of Kaempferia parviflora from in vitro cultured plantlets [28], tissue cultures of six genotypes of Deschampsia Antarctica [29], biomass of in vitro shoots and roots of Eryngium species (E. campestre, E. maritimum, E. planum) [30] or leaves from micropropagated plantlets of Passiflora setacea cv BRS Pérola do Cerrado [31].…”

Galic acid as a non-specific regulator of phenol synthesis and growth of regenerate plants of Corylus avellana (L.) H. Karst. and Salix alba L. in vitro

“…The volume ratio of solvent to raw mass of leaves was 1/10 (v/v). The total content of phenols in the leaves was determined using spectrophotometry with Folin-Ciocalteu reagent [31]. The calibration graph was plotted using gallic acid.…”