Production of Picroside II from Picrorhiza kurroa Royle ex. Benth in vitro cultures under predisposition of an endophyte
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Date
2018-07
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G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand)
Abstract
Biological resources have been used as vital resource for existence of life for ages These are potent source of pharmacotherapeutics widely used in Naturopathy, Ayurveda, Unani, siddha and Homeopathy since time immemorial. Picrorhiza kurroa Royle ex Benth, a therapeutic perennial herb, primarily distributed in the north-west Himalayan Mountains at altitudes ranging from 3000- 5000m above msl, is a representative endemic to alpine Himalayas. The plant has been utilized in formulation of various drugs commercially due to presence of monoterpene iridoid derived secondary metabolites such as picrosides which include picroside-I, picroside-II and other metabolites. Reckless collection from wild and lack of planned cultivation of the plant has threatened its status in wild and has been listed as 'endangered' species by IUCN. It is essential to apply biotechnological approaches to enhance the production of picrosides in vitro level to ensure its proper regeneration and constant supply. In the current investigation, an endeavor has been made to assess the quantity of Picrosides content and expression analysis of biosynthetic pathway genes through elicitation method in suspended callus regenerated from leaf explants in vitro. MS media supplemented with 0.75 mg/L TDZ + 0.75 mg/L IBA was found effective in callus induction. The effect of endophytic fungus Fusarium sp. L4 (KU180458) culture as elicitor was used to recognize some facts about the expression of biogenesis pathway genes (HMGR, DXS, CAM) and picroside-II content. Elicitor study results indicated that it might be possible that IPP for biogenesis of Picroside(s) is supplied by mevalonate, non mevalonate and phenyl-propanoid pathways. Autoclaved and filtered extracts were used for elicitation to know the possible better method of elicitation in order to enhance picrosides production. The use of fungal culture as elicitor was effective in enhancing the secondary metabolite in Picrorhiza kurroa Royle ex Benth especially Autoclaved extract (AE) as compared to Filtered Extract (FE). Correlation study between expression analysis of putative gene(s) and picroside-II content was also carried out. Results obtained from above investigation can be summarized as that in vitro elicited callus culture of Picrorhiza kurroa Royle ex Benth can be used for large scale production of Picroside-II, an immensely valuable Bioactive compound.
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