Characterization and comparison of bioactive constituents in the in vitro regenerated and wild roots of Himalayan medicinal herb Valeriana jatamansi Jones
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Date
2018-08
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G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand)
Abstract
The medicinal herb Valeriana jatamansi Jones is a native of Northeastern Himalayan region belonging to family Valerianaceae and growing at altitudes of 1500-3000 m asl. The in vitro direct-, indirect rhizogenesis
and plantlet regeneration was achieved from leaf explant in MS medium supplemented with ppm quantities of NAA (2.0)+BAP (2.0); IBA (1.0)+NAA (1.0)+BAP (0.1) and BAP (2.0)+NAA (0.2), respectively. Multiple shoots and plantlet regeneration was achieved from nodal explant in MS medium supplemented with ppm quantities of NAA(0.1)+TDZ(1.0) and IBA(1.0)+AgNO3(1.0), respectively. Phytochemical screening in crude
and methanolic root extract by FTIR confirmed the presence of alkaloids, saponins, tannins, flavonoids, steroids and terpenes, and polyphenols. Methanol was the best solvent followed by dichloromethane and ethylacetate for extraction of phytochemicals from root and leaf of wild plant. The highest phytochemicals like phenol (351.0mg GAE g-1 extract), flavonoids (84.0mg QE g-1 extract), tannin (814.0mg TAE g-1 extract) and orthodihydroxy phenol (47.0mg CE g-1 extract) content and antioxidant activities like total antioxidant (935.4 AAE g- 1), FRAP (810.1 AAE g-1 extract), DPPH radical scavenging (IC 50=11.8μg ml-1), Superoxide radical scavenging (IC50=18.4μg ml-1), Hydroxyl radical scavenging (IC50=18.1μg ml-1) and Metal ion chelating (IC50=11.5μg ml-1) were found to be highest in methanolic extract of 6 months grown wild roots at 35ºC. The methanolic extracts of 6 months grown roots at 35ºC showed lowest MIC (62.50 μg) and maximum efficacy at par to gentamycin standard against 6 human pathogenic strains i.e., S. typhaemurium-52, S. typhaemurium-149, S. entiritidis-ATCC 13076, S. typhaemurium-ATCC 51812, E. coli-ATCC-25922 and E coli strain-13. Effect of maturity of roots (in vitro- and wild grown) and thermal stress indicated that the phytochemical, antioxidant, antioxidant enzymes and antimicrobial activities in 6 months grown in vitro roots were comparable to 3 months grown wild roots indicating adaptation towards higher temperature tolerance, since V. jatamansi was a temperate herb. Micronutrient analysis showed that Fe (0.254 ppm) was found to be significantly highest than Zn (0.137 ppm) in wild leaf and root. The third largest element in root was Mn (0.052 ppm) and Ca (0.118 ppm) in wild leaf because Mn facilitated the uptake of other nutrients like Fe and inorganic phosphate in root and Ca acted as binding agent in cell wall fusion in leaf. GC-MS analyses identified 38 constituents comprising 78.33% of total compositions of which 28% were contributed by patchouli alcohol confirming V. jatamansi as a patchouli chemotype. Valerenic acid content quantified by HPLC in the in vitro and wild roots was 8.6 mg g-1 and 25.09 mg g-1 dry weight, respectively.
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