Callus formation, phenolics content and related antioxidant activities in tissue culture of a medicinal plant colocynth (Citrullus colocynthis)

Abstract

Callus cultures from stems, leaves and roots of colocynth were initiated on MS media supplemented with various combinations of 2,4 dichlorophenoxyacetic acid (2,4-D) with kinetin (KIN) and benzyladenine (BA) with α-naphthaleneacetic acid (NAA). The highest percentage of callus formation frequency (98.9%) was obtained from stem explants grown on MS media supplemented with (1.0 mg/L) 2,4-D + (1.0 mg/L) KIN. The total phenolics and flavonoid content of the colocynth callus cultures were measured. The results showed that the MS medium supplemented with 6.0 mg/L 2,4-D + 2.0 mg/L KIN (MD3) gave the highest content of total phenolics (19.2 mg/100g d.w.) in leaf-derived calli. The highest content of flavonoids (47.3 mg/100g d.w.) was obtained in stem derived calli grown on the same medium (MD3). Antioxidant activities of extracts were determined using different assays, including DPPH radical scavenging activity, hydrogen peroxide (H2O2) scavenging activity and ferric reducing power. Leaf-derived calli cultured on MS medium + 2.0 mg/L 2,4-D + 1.0 mg/L KIN (MD1) showed the highest DPPH radical scavenging activity (85.3%). The highest percentage of H2O2 scavenging activity (61.4%) was detected in leaf explant-derived calli growing on MD1. The leaf-derived calli growing on (MD3) gave the highest ferric reducing power (22.3 μg/g d.w.), compared to the activities of stems, leaves and roots of in vitro grown seedlings (3.28, 12.9 and 2.85 μg/g d.w.), which were used as controls. On the basis of the current findings, we conclude that MS media supplemented with different combinations of 2,4-D and KIN yields higher phenolics, flavonoids contents and antioxidant activities than MS media supplemented with BA and NAA.