These include superoxide radicals (O2 −), singlet oxygen (1O2), hydrogen peroxide (H2O2) and hydroxyl radicals (OH ) which causes tissue injury. These are highly reactive species and can seriously disrupt normal metabolism through oxidative damage to membrane lipids, protein pigments and nucleic acid and ultimately results in cell death. To counter the hazardous effect of reactive oxygen species under stress, plants have developed or have evolved a complex antioxidative

defense mechanism system which involves both enzymatic and non-enzymatic metabolites antioxidant such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) which are efficient antioxidant enzymes. The antioxidant metabolism is enhanced during differentiation in vitro, and antioxidant profiles also vary throughout different phases of culture [6]. The production RG7422 datasheet of ROS has been associated

with plant selleck recalcitrance during in vitro culture [7]. In this work, we also match up the altered levels of antioxidant enzymes produced during the culture conditions with those of ex vitro regenerated plants and their part in thriving plant to external environmental conditions. Seeds of C. halicacabum were collected from the plants growing in the botanical garden of the university. The seeds were washed thoroughly under running tap water for 30 min followed by treatment with 5% (v/v) Labolene, a liquid detergent for 15 min. The seeds were then rinsed thoroughly and treated with 0.1% (w/v) HgCl2 for 5 min. After rinsing 5–6 times with sterilized distilled water, the seeds were inoculated in Murashige and Skoog’s medium [8] for germination. Hypocotyl segments excised from

Adenosine triphosphate 7 days old aseptic seedling were used as an explant. MS medium supplemented with 3% (w/v) sucrose and 0.8% (w/v) agar or 0.25% (w/v) gelrite was used during the investigation. The pH of the medium was adjusted to 5.8 with 1 N NaOH or HCl prior to autoclaving. The media were dispensed in 25 mm × 150 mm test tubes (Borosil, India) each containing 20 ml of medium and cotton plugs (single layered cheese cloth stuffed with non-absorbent cotton) were used as closures. Glasswares, culture media, and instruments were sterilized by autoclaving at 121 °C at ∼105 kPa for 20 min. All the cultures were maintained at 24 ± 2 °C under 16 h photoperiod with a photosynthetic photon flux density (PPFD) of 50 μmol m−2 s−1 provided by 40 W cool white fluorescent lamps (Philips, India) and with 60–65% relative humidity. For multiple shoot induction, excised hypocotyl explants were inoculated on MS medium augmented with various cytokinins, BA (0.5, 2.5, 5.0, 7.5, and 10.0 μM) and TDZ at lower concentrations (0.1, 0.3, 0.5, 0.7, and 0.9 μM) individually. Initially, cultures were subcultured onto the same fresh medium after every 2 weeks resulted in fascinated, distorted, stunted, and clumped shoots which did not elongate further.