Phytochemical Screening of Callus and Cell Suspensions Cultures of Thevetia peruviana

Abstract

Thevetia peruviana is an ornamental shrub grown-up in many tropical region of the world. This plant produces secondary metabolites with biological properties of interest for the pharmaceutical industry. The objective was to determine the secondary metabolites profile of callus and cell suspension cultures of T. peruviana and compare them with those from explant (fruit pulp). Extracts in 50% aqueous ethanol and ethyl acetate were prepared. The phytochemical analysis was performed using standard chemical tests and thin layer chromatography. In addition, total phenolic and flavonoids compounds (TPC and TFC), total cardiac glycosides (TCG) and total antioxidant activity (TAA) was determined during the cell suspension growth. Phenolic chemical profile was also analyzed by high performance liquid chromatography (HPLC). Common metabolites (alkaloids, amino acids, antioxidants, cardiac glycosides, leucoanthocyanidins, flavonoids, phenols, sugars and triterpenes) were detected in all samples. The maximum production of extracellular TCG, TPC, TFC and TAA in cells suspensions were at 6-12 days; in contrast, intracellular content was relatively constant during the exponential grown phase (0 to 12-days). HPLC analysis detected one compound with retention time at 11.6 min; this compound was tentatively identified as dihydroquercetin, a flavonoid with anti-cancer properties. These results provide evidence on the utility of the in vitro cell cultures of T. peruviana for valuable pharmaceutical compounds production. HIGHLIGHTS Callus and cell suspensions of T. peruviana have similar phytochemical profile to in vivo plant. Cell culture of T. peruviana is a reliable platform for high-value metabolites production. Cardiac glycosides and phenolic are the most valuable metabolites detected in plant cell cultures  Dihydroquercetin production in a free and conjugated form in cell cultures is highlighted 2 Mendoza, D.; et al.

Optimization of Flavonoid Production in Plant Cell Culture of Thevetia peruviana Elicited with Methyl Jasmonate and Salicylic Acid

“…Researches conducted by Arias and coauthors [25] and Tabata [26] in in vitro cultures of T. peruviana reported the increase of cardiotonic glycosides and peruvosides using MeJa and SA elicitors. A recent study published by Mendoza and coauthors [7] reported cardiac glycosides, phenolic, and flavonoids compounds in cell suspension cultures of T. peruviana during the exponential grown phase. Considering that secondary metabolites are of pharmaceutical interest, it is necessary to explore ways of large-scale production at industrial levels, which have already been achieved with plants such as Taxus sp.…”

“…The explants were extracted from fruit pulp and put aseptically in sterile solid Schenk & Hildebrandt (SH) medium [30], supplemented with 2 mg L -1 2,4-dichlorophenoxyacetic acid (2,4-D), 0.5 mg L -1 kinetin, 30 g L -1 sucrose, 7 g L -1 agar and 1 g L -1 myoinositol (pH 5.8), sterilized at 121°C and 1 atm during 20 min. Callus cultures were kept in photoperiod of light 12 hdarkness 12 h. Sub-cultures were made every three weeks until obtaining friable callus [7].…”

“…Production of terpene and phenolic type compounds has been reported in leaves [4], bark [5], flowers [6], fruits [7] and seeds [8] of T. peruviana; in the latter group of metabolites stand out the flavonoids kaempferol and quercetin, which have three rings in their structure, because they derive from aromatic amino acids such as phenylalanine (Phe) and tyrosine (Tyr) [9]. These compounds are produced by acetate/malonate, shikimic acid, pentose phosphate metabolic pathway [10], and through metabolism of phenylpropanoids [11].…”

Inhibition of melanogenesis by Aster yomena callus pellet extract in melanoma cells and patients with skin pigmentation

“…In particular, metabolites (including alkaloids, flavonoids, polyphenols, terpenes, triterpenes, cardiac glycosides, paclitaxel and related taxoids) with various physiologically active functions can be produced using this method 14 , 15 . Therefore, the plant tissue culture technique is actively studied to continuously improve the production of biologically valuable high-quality materials 16 - 18 . Nonetheless, research on the selection and function of plant callus materials that can be used as functional cosmetic ingredients is still lacking.…”

Determination of the Qualitative Composition of Biologically Active Substances of Extracts of In Vitro Callus, Cell Suspension, and Root Cultures of the Medicinal Plant Rhaponticum carthamoides

“…The study and research of alternative methods of obtaining compounds with useful and unique characteristics from plant materials is relevant. The biosynthesis of in vitro transformed callus, cell suspension, and root cultures (hairy roots), from the point of view of the controlled production of biologically active substances with desired properties, has potential, and is of interest to both researchers and producers [25].…”