Colchicine Induced Mutation in Nigella sativa Plant for the Assessment of Morpho-Physiological and Biochemical Parameter Vis-A-Vis In Vitro Anti-Inflammatory Activity

Abstract

Background: Nigella sativa (NS), an herbaceous medicinal plant recognized for its diverse beneficial applications as a spice and traditional medicine. Objective: The present study was targeted to explore the antioxidant potential of Nigella sativa in response to colchicine-induced mutation. The stress condition brought due to mutation may affect the medicinal value (anti-inflammatory activity) of the plant. Method: Nigella sativa seeds were imperiled to colchicine treatment at various concentrations viz. 0.00625, 0.0125, 0.025, 0.05 and 0.1% subjected for analysis. Result: The colchicine treated plant (polyploid/ mutant) at 0.025% concentrations showed significant variation at morpho-physiological and biochemical level with respect to control (p value < 0.05). At the morphological level, the plant showed enlargement of shoot length (33.760±2.295mm), root length (13.546±1.535 mm), and leaf area (22.836±1.524 mm2). The analysis of seeds showed enhanced seeds per pod (49.333±4.163), weight of seeds (2.810±0.010g), length (3.133±0.089mm), and width (1.123±0.044mm) when compared with control. The physiological parameters also showed significant enhancement for stomatal index (35.456±4.751%), chlorophyll A (9.053±0.865 µg/gfw), chlorophyll B (4.990±0.763 µg/gfw), and total carotene content (773.190±5.906 µg/gfw). However, the fresh weight/ dry weight ratio (10.738±3.031) was found to be deprived. Furthermore, biochemical parameters viz. total flavonoid (seeds 1.973±0.134; plant 1.703 ± 0.064 mg eqv QE/g of tissue), total phenolic (seeds 15.690±1.495; plant 8.220±0.070 mg eqv GA/g of tissue), total carotene (seeds nil; plant 773.190±5.906 µg/gfw), and total antioxidant (seeds 0.445±0.102; plant 0.386±0.010 mM eqv AA/g tissue) were significantly elevated at 0.025% of colchicine treatment. When the in vitro anti-inflammatory activity was targeted, a significant escalation was observed for inhibition of albumin denaturation (97.466±2.835%), proteinase inhibitory activity (62.290±6.475%), heat-induced hemolysis (89.873±3.533%), hypotonicity induced hemolysis (92.572±3.527%), anti-lipoxygenase activity (96.010±3.098%), and cyclooxygenase inhibitory activity (68.296±3.920%) at 500µg/mL concentration of extract. Conclusion: Thus, it can be concluded that 0.025% of colchicine can induce significant (p value < 0.05) mutation in the Nigella sativa plant, which may lead to alterations at morpho-physiological and biochemical levels. Such treatment induces stress in the plant and leads to elevated antioxidant levels. This in turn elevates the therapeutic potential of the plant. Hence, our study is a novel and open-ended finding to explore various other medical properties of the plant with respect to colchicine-induced mutation.