Comparative analysis of withanolide accumulation in leaves and calli of Withania somnifera (L.) Dunal grown in different soil provenances
摘要
Withania somnifera (L.) Dunal (Indian ginseng); a valuable plant of Ayurvedic Medical System is helpful in treating diverse health issues. It possesses a number of medicinally important secondary metabolites such as withanolides, which are probably synthesized under stress conditions. However, its restricted distribution among plants and limited yield in W. somnifera itself, coupled with declining availability due to urbanization, unsustainable harvesting and habitat loss, pose significant challenges. These constraints highlight the urgent need to explore alternative strategies for producing such medicinally valuable metabolites in higher quantities. The current investigation was carried to upscale in vivo and in vitro synthesis of withanolides. For this purpose, a range of soil provenances, including the polluted site, were used to grow seedlings (1-month old) of Withania in pots as well as in the field. The calli were produced on basal MS medium comprising various combinations of BAP and NAA. Leaf explant from pot plants containing soil of selected provenances were used to find out the effect of soil provenances and hormonal combinations on the biosynthesis of withanolides in calli. Total alkaloids were extracted from different plant parts and calli and estimated for withanolides and withaferin-A using HPLC. Interestingly, total withanolide contents were recorded to be the highest in the callus raised from BEE (Before Effluent Entry) leaf explant on MS medium comprising BAP (1.0 mg/L) and NAA 0.5 mg/L) and sub-cultured on MS medium with BAP (2.0 mg/L) and NAA (0.1 mg/L) as compared to all other calli including those raised by the use of leaf explant from IGS (Irrigated Garden Soil) and DSS (Ditch Site Soil) cultivated pot plants. Additionally, amount of withanolides in the calli grown from DSS and BEE leaf explants on BAP (1.0 mg/L) with NAA (0.5 mg/L) and sub-cultured on BAP (2.0 mg/L) with NAA (0.1 mg/L) was found to be 153% and 1050% higher, respectively, as compared to corresponding in vivo leaves. The current investigation clearly indicates that the physiological condition of plants growing in selected/treated or polluted soil promotes in vitro withanolides production and accumulation in the callus of Withania somnifera (L.) Dunal.