Influence of photoperiod on growth, bioactive compounds and antioxidant activity in callus cultures of Basella rubra L

Basella rubra L. is an important green leafy vegetable vine and is known for its health benefits in traditional medicine. Light is a basic physical factor essential to the development and bioactive secondary metabolite production in in vitro callus cultures. The present study researched the impact of different photoperiods on biomass, bioactive compounds, and antioxidant activity in callus cultures of B. rubra. The in vitro seedling based cotyledonary leaf explants responded differently, when cultured on Murashige and Skoog (MS) medium with varying concentrations and combination of auxins and cytokinins. The best callus proliferation was found in MS medium with 0.1 mg.L^-1 1-naphthaleneacetic acid (NAA) and 6 mg.L^-1 6-benzylaminopurine (BAP), with greenish callus inception by about 2 weeks. The growth curve recorded for 6 weeks of culturing revealed that the photoperiod effect was found to be pivotal for acquiring biomass. At the fifth week, the continuous light supported maximum biomass (12.42 g) production followed by the 16:8 h photoperiod (9.02 g) and continuous darkness (4.28 g). The 80% ethanol extract of 1-week-old callus that grows under the 16:8 h photoperiod showed the highest total phenolic content (TPC) (74 mg.100 g^-1 fresh weight, FW) when compared to all other extracts at different stages. The ferric reducing antioxidant power assay showed the highest (336.23 mg.100 g^-1 FW) activity in methanol extractions of first-week callus cultures maintained in the continuous light condition. HPLC-UV identification and quantification of individual phenolics and flavonoids, such as gallic, trans-cinnamic, quercetin, protocatechuic and rutin, were highest in the callus cultures. The outcome of this study is significant to this plant, as B. rubra is familiar for its important health constituents with high-value bioactives and applications in the pharma and nutraceutical industries.

Keywords: Antioxidant activity; Basella rubra; Callus cultures; Growth curve; Phenolic acids; Photoperiod.