Influence of plant growth regulators and chitosan elicitation oncallus induction and improvement of morphophysiological, metabolic andantioxidant traits in Chrysanthemum indicum L.
DOI:
https://doi.org/10.24154/jhs.v20i2.4515Keywords:
Antioxidant activity, chitosan, Chrysanthemum, elicitor, in vitro cultureAbstract
Chitosan, as a natural elicitor, can enhance plant defense mechanisms and modulate physiological, biochemical, metabolic, and antioxidant pathways. Its application in vitro offers a promising strategy to improve the production of valuable secondary metabolites in Chrysanthemum indicum L. In this study, Murashige and Skoog (MS) medium was supplemented with various hormonal combinations viz., H1 (control), H2 (1 mgL-1 NAA + 2 mgL-1 BAP), H3 (0.2 mgL-1 2,4-D), H4 (0.2 mgL-1 BAP + 0.5 mgL-1 2,4-D), and H5 (2 mgL-1 BAP + 2 mgL-1 2,4-D) and applied to different explants (leaf and stem) to optimize callus induction. Optimization was performed by evaluating callus induction percentage, initiation time, fresh weights, and callus volume. Following chitosan elicitation, several metabolic and biochemical traits were evaluated, including carotenoids, total phenolics, flavonoids, anthocyanins, proline content, antioxidant capacity, and the enzymatic activities of catalase and peroxidase. Among the treatments, the combination of 2 mg/L BAP + 2 mg/L 2,4-D applied to leaf explants under light conditions, produced the highest callus induction rate (96.66%), minimum days to response, and significantly increased fresh weights and callus volume. Chitosan treatments (25, 50, and75 mgL-1) were applied to calluses derived from the optimized treatment (2 mg/L BAP+2 mg/L 2,4-D applied to leaf explants). Among the tested concentrations, chitosan at 50 mgL-1 was the most effective, leading to the maximum increases in fresh weight (2.63 g), callus volume (1.81 mm³), carotenoids (2.93 mg/g FW), total phenolics (67.40 mg/g FW), flavonoids (41.32 mg/g FW), and antioxidant activity (62.75%). Additionally, this treatment enhanced anthocyanins (5.19 mg/g FW), proline (6.14 mg/g FW), catalase (14.37 U/mg FW), and peroxidase (2.45 U/mg FW) activities. Overall, the combined use of plant growth regulators (BAP+2,4-D) and chitosan, particularly at 50 mgL-1, presents an efficient approach to enhance callus development and improve physiological, metabolic, and antioxidant responses in Chrysanthemum indicum L. under in vitro conditions.
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