Metabolite profiling of in vitro established cultures of Picrorhiza kurroa Royle ex Benth in different growth regime(s)
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Date
2017-08
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G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand)
Abstract
The herbs as medicine are being traditionally in practice from ancient time for healthcare to safeguard humanity. These are being used in Ayurved, Unnani, Siddha and Homeopathy with different kind of healing practices in animals as well as human being with different drug delivery systems based on active ingridients.Picrorhiza kurroa Royle ex Benth, a Scrophulariaceae member, is a reputed and highly studied medicinal herb endemic to alpine and sub alpine belts of the Himalayas. Several monoterpenoid glycosides are present which are known as picroside I, picroside II, picroside III, more recently picroside IV are being studied specially for hepatomodulator.The plant has been overconsumed for its valuable glycosides, hence now it comes under endangered category according to Red Data Book. The depleting strategies of the herb in the wild which requires implementation of conservation and sustainable utilization practices. It is essential to apply biotechnological approaches including tissue culture which in turn will also help for the defined conditions to enhance the production of picrosides in vitro. In the present research work, an attempt had been made to assess the quality/quantity of Picroside content, total protein profiling and study of SOD pattern analyses through use of elicitors and phytoregulators. The suspended leaves regenerated from in vitro Picrorhiza kurroa cultures using leaves as explant(s) from germplasm evaluation facility. Different MS supplemented with 0.75 mg/L TDZ + 0.75 mg/L IBA was used as callusing medium and BAP 0.50 mg/L + Kinetin 0.75 mg/L was used for shoot proliferation.GA3 as a Hormonal elicitor was used as elicitor to recognize some facts about the expression of biogenesis pathway protein and the content of picroside-II. Results of elicitor study revealed that it might be speculated that mevalonate/ non mevalonate pathways are being involved in Picroside(s) biogenesis. GA3 downregulates the biosynthesis of picrosides as it also follows the same biosynthetic pathways. Electrophoresis of protein reveals bands like 84,67,58,37,25 kDa which were observed in all samples taken at different days interval while other bands were seen in 12 and 18 days samples harvested from suspension cultures, which implies that some proteins are produced under the increased concentration of GA3. Achromatic regions on the native gel indicate the activity of SOD. They play important roles throughout plant growth and development in vitro. SOD activity was found more where higher concentration of GA3 than differently treated samples Although the exact mechanism is to be explored. These results indicative of the increased amount of GA3 indicative of the picrosides biosynthetic pathways.
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