In vitro morphogenesis of Polygonatum verticillatum (L.) all along with comparative study of biological activities of rhizomes collected from different altitudes of Uttarakhand
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Date
2021-02
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G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand)
Abstract
Medicinal plants are the valuable and harmless source of drugs that have drawn the attention of many scientists and pharmaceutical companies for the preparation of many natural products. Polygonatum verticillatum (L.) All., an Asparagaceae family member, medicinal herb of North West Himalaya. It is also a member of Astavarga group of medicinal plants that have high medicinal value in Ayurveda. Regeneration of this plant through seeds is slow process as the seeds have double dormancy. So the present study is carried out for the rapid propagation of these medicinal plants through the tissue culture. Maximum per cent callus induction was found in MS medium supplemented with TDZ (0.5 mg L-1) and 2, 4 –D (1.0 mg L-1). Maximum per cent (81.25 ± 3.6) shoot induction was achieved in MS fortified with mT (0.5 mg L-1) with maximum (23.67 ± 2.02) no. of shoots/explant. Maximum per cent (71.33 ± 8.3) root induction was observed in MS medium supplemented with IBA (1.0 mg L-1) and maximum no. of roots/explants (10.33 ± 0.6) was found in MS medium fortified with NAA (1.0 mg L-1) + Kn (0.5 mg L-1). Maximum per cent embryo induction (62.5 ± 7.2) was achieved in MS containing Kn (2.0 mg L-1) + NAA (1.0 mg L-1). Synthetic seeds were prepared in 2.5 % sodium alginate and 2 % CaCl2. In the present study, the effect of altitudes on the antioxidant activity, Total flavonoid content and total phenol content was also evaluated in aqueous and methanol extracts of rhizomes of P. verticillatum collected from different altitudinal regions. The rhizomes were collected from Chamoli (P1), Bhowali (P2), Jageshwar (P3), Munsiyari (P4). Maximum extraction yield (%) was achieved in P4 (14.78 %) and P4 population (12.43 %) in aqueous and methanol extract respectively. Majority of phytochemicals were present in aqueous and methanol extract except tannins and alkaloids were absent in aqueous solvent. DPPH scavenging activity (%) was maximum in P4 (38.34) and P3 (32.368) in aqueous and methanol extract respectively. Comparatively lower IC50 (μg/ml) values of DPPH were obtained in P4 population (172.17) in aqueous and P3 popualtion (183.71) in methanol extract. Maximum FRAP (μg BHT/mg) activity was observed in P3 (77.21) and P4 population (36.69) in aqueous and methanol extract respectively. Total antioxidant activity (μg VCE/mg) was highest in P4 population (23.35) in aqueous extract and P3 population (16.87) in methanol extract respectively. Maximum flavonoid content (μg QE/mg) was achieved in P3 population (17.67) in aqueous extract and P4 population (24.17) in methanol extract respectively. Highest total phenol content was achieved in P4 population (35.76) and P4 popualtion (23.83) in aqueous and methanol extracts respectively. The GC-MS analysis of aqueous and methanol extracts of P. verticillatum rhizome showed the presence of 27 and 30 bioactive compounds respectively. In aqueous extract 7-Nonenamide (22.19%), 5- Hydromethylfufural (8.87 %), 3-Furaldehyde (8.82 %), 5-Hydromethylfurfural (8.87%), D- Alanine, Npropargyloxycarbony - isohexyl, ester (7.20 %),2,4- Dihydroxy-2,5-dimethyl-3(2H)- furan-3-one (5.76 %), 2-acetyl – 1-pyrroline (5.40 %), 3- Amino-2- oxazolidinone (4.39 %), 1,3 –Dioxolane, 2,4,5- trimethyl- (3.77 %) were reported as main compounds were the major compounds while in methanol extract 4-Hexene-3-one, 4, 5 – dimethyl (40.60 %), furfural (26.07), (R) – (-) – Methyl -2- butane (4.20 %), Cholesterol (3.06%), 10, 11, 12, 13, 14, 15, 16, 17-tetradecahydro-1H-cyclop enta [a]phenanthren-3-ol (2.86 %), 26-Nor-5-cholesten-3á-ol-25- one (2.47 %) were the major compounds were the major compounds. Utilizing the findings of the present research not only provides a method for mass propagation of the species but also helps in assessing the phytochemical differences in the populations growing at different altitudes.