Rapid in vitro callogenesis and phytochemical screening of leaf, stem and leaf callus of mussaenda frondosa linn.: A medicinal plant

Abstract

Objective: To standardize the protocol for rapid callogenesis in Mussaenda frondosa L. using leaf explants. Qualitative and quantitative phytochemical analysis of leaf, stem, and callus cultures. Methods: The leaf explants were inoculated onto Murashige and Skoog medium supplemented with varying concentrations of growth regulators such as 2, 4-D, NAA, benzylaminopurine, Kn for the induction of callus. Qualitative and quantitative analysis of total phenol, flavonoid and alkaloid contents of leaf, stem, and callus were tested by standard methods. The antioxidant activities were investigated using 1, 1- diphenyl-2-picryl hydrazyl radical scavenging method and reducing power assay. The anti-inflammatory activity was evaluated by membrane stabilizing activity. Results: Pale green, healthy, friable, and fast growing callus were obtained on the medium enriched with NAA (2 mg/l)+Kn (4 mg/l). Quantitative determination showed the highest concentration of total phenolics in the methanolic extract of in vitro grown callus (10±1.1 mg of GA/g of extract), flavonoids in methanolic stem extract (137±1.6 mg of quercitin/g of extract) and alkaloids in methanolic extract of leaf (118.3±1.5 mg/10 g of extract). The methanolic leaf extract exhibited the highest free radical scavenging activity with an inhibitory concentration 50% (IC50) value of 40.6±10.06 μg/ml. The highest membrane stabilizing activity was shown by chloroform extract of the leaf (66.02%). Conclusion: This preliminary phytochemical and pharmacological analysis may form the basis for drug development in future using callus cultures of M. frondosa.