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Open Access
Biochemical and molecular attributes of the diverse genetic pool of Brassica juncea var. rugosa (pahari rai) for identification of elite germplasm having potent bioactivity
(G. B. Pant University of Agriculture and Technology, Pantnagar, 2022-09) Adhikari, Ankur; Punetha, Himanshu
Brassica juncea var. rugosa (pahari rai), a broad leafy cruciferous vegetable is an essential and easy source of nutrition with numerous health-promoting factors. Eighteen different germplasms of Pahari rai and two checks (Pusa saag and Kranti) have been cultivated during seasons 2019-20 and 2020-21. Germplasm had totally different selected agro-morphological traits. PCA analysis depicted that PC1 (35.25%), PC2 (18.71%) and PC3 (17.09%) contributed to the morphological variability among the germplasms for the traits under study. Tocher method divided all germplasms into four different clusters as per morphological characters. A biplot between PC1 & PC2 showed, germplasm IC-399826 is within the ace of origin were more stable and less diverse, whereas IC-597821, IC- 298019 and IC-276011 are away from the origin, which were less stable and more diverse. Genetic diversity and population structure of cultivated germplasms using 100 SSR primers subjected to Darwin 5 software. A total of 74% SSRs resulted in polymorphic amplicons. UNJ-based dendrogram grouped all the 18 germplasm along with two checks into two distinct clusters. PCA of genetic diversity showed that the first and second principal components accounted for 29.25% and 14.7% of the total variation. Phytochemical and antioxidant activity of leaf extracts showed the highest phenolic (6.45±0.045 mg GAE g-1), flavonoid (6.32±0.06 mg QE g-1), ODP (4.66 ± 0.52 mg CE g-1) and tannin (3.46 ± 0.02 mg tannic acid g-1) content were observed in the EEC-25 leaf methanolic extract. The highest protein content in EEC-25 (400.97±16.5 μg QE g-1) followed by IC-524259 (380.09±17.22 μg QE g-1) and IC-350800 (379.23±18.26 μg QE g-1). The foremost concentration of carbohydrates was observed in germplasms IC-363758 (31.47±0.31 mg glucose g-1), IC-410471 (31.12±0.18 mg glucose g-1) and EEC-25 (26.45±0.19 mg glucose g-1). In-vitro antioxidant potential viz., total antioxidant content was observed highest in EEC-25 methanolic leaf extract (41.91±0.28 mg AAE g-1), FRAP activity in EEC-25 (5.91±0.68 mg AAE g-1), maximum DPPH free radical scavenging activity in IC- 597933 (IC50=21.45±3.4 μg mL-1), ABTS free radical scavenging activity in IC-524259 (94.17±0.48 μg mL-1), superoxide radical scavenging activity of methanolic extract of PR-15 (IC50=124.37 μg mL-1), highest metal ion chelating activity of EEC-25 methanolic extract (IC50=52.22 μg mL-1), maximum reducing power activity in EEC-25 and IC-399839, all being significantly different (p≤0.05) from each other. Positive correlations have been observed among the total antioxidant, total phenolic, total flavonoid content, ODP, DPPH and FRAP. Pahari rai was observed to be a rich source of MUFA and PUFA. An essential metric for assessing the nutritional value of various edible oils is the ω6:ω3 ratio was observed to be maximum in IC-350800 (2.53), IC-338535 (2.34) and IC-410471 (2.19). Micronutrient analysis Pahari rai leaf observed to be the rich source of essential minerals such as Mn, Fe, Zn, Ca, Cu and Mg. The anti-inflammatory activity among the elite accessions the IC-350800 (IC50 =25.19±1.06 μg mL-1) was observed to be the maximum. No zone of inhibition was evident against selected human pathogenic bacteria using methanolic leaf extracts of elite accessions of Pahari rai. Invitro cytotoxicity in MDBK and Vero against the selected methanolic leaf extracts (IC-524259, EEC- 25, IC-597933, IC-350800 and Pusa saag) was non-significant. Results clearly revealed the presence of significant amounts of phytochemicals, antioxidant activities and in vitro anti-inflammatory activities in leaves of Pahari rai. The multiutility of this leafy vegetables against numerous pathological ailments was attributed to pharmacological activities of phytochemicals and for the development of functional food for human consumption.
Open Access
Studies on starch metabolizing enzymes and physico-chemical methods to modify wheat flour for high resistant starch
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2022-03) Jadhav, Reshma Ravindra; Agrawal, Sanjeev
The present investigation was carried out to study the effect of key starch metabolizing enzymes on resistant starch accumulation during grain filling stages and to modify the wheat flour for high resistant starch using different physico-chemical methods. The activities of starch metabolizing enzymes viz. AGPase, SSS, SBE, GBSS and DBE were determined during grain filling stages at S1, S2, S3, S4 and S5 stages (7, 14, 21, 28 and 35 DAA, respectively) in wheat genotypes.The activities of AGPase, SSS and SBE weresignificantly higher in UP-2785, UP-262 and UP-2584 than the other genotypes. These genotypes also showed comparatively higher grain weight (TKW), total starch and amylopectin accumulation, hence these enzymes play important role in regulating the kernel weight at grain filling stage. Genotypes UP-2748, UP-2903 and UP- 2584 were observed to be higher inaverage GBSS and DBE activities,amylose and resistant starch content. These enzymes may play important role in amylose and resistant starch accumulation. Higher fold increase in AGPase, SSS SBE activities and total starch, amylopectin accumulation and TKW was observed at stage, S2(14DAA), whereas higher fold increase in GBSS and DBE activities, amylose and resistant starch accumulation was observed at S3 stage (21DAA). So, duration of these twostages S2 and S3 may be the important for maintaining kernel weight, starch quality and quantity. AGPase, SSS and SBE enzymes were positively and significantly correlated with total starch, amylopectin and TKW, whereas GBSS and DBE enzymes were positively and significantly correlated with amylose and resistant starch accumulation. After the harvesting, genotypes UP-2309, UP -262 and UP-2748 were screened among the nine genotypes on the basis of different characters like chapatti making, iron, zinc content and resistant starch content and other quality parameters. These three genotypes were irradiation with gamma rays with different intensity, which affects positively in increasing amylose and resistant starch content. Genotype UP-262 was found to have comparatively higherpercent of increased resistant starch content followed by UP-2903 and UP-2748 due to irradiation treatment. The treatment 5 and 7 KGy showed the better performance for high resistant starch content compared to other treatments. The maximum decreases in transmittance was observed at 5KGy (UP-2478) and7 KGy (UP-262 and UP-2748) at 1047 and 1022 cm-1 spectra, which was important to be noticed when compared to the control (non irradiated). The results thus point out strong molecular reorganization in 5 and 7 KGy irradiated wheat flour compared to control. Wheat flour fortified with chickpea, finger millet or soybean showed high resistant starch and low glycemic index. Highest value of resistant starch content was observed in wheat–soybean blend i.e soybean blend showed better performance followed by chickpea and finger millet.
Open Access
Comparative phytochemical analysis of Berberis lycium royle from different regions of Uttarakhand and its pharmacological potential in combination with Withania somnifera
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2022-03) Shruti; Verma, A.K.
Plant samples of Berberis lycium Royle (BLR) were collected from different Himalayan regions of Uttarakhand which ranged from 533 to 1922 m/ASL altitude. The bioactive compound berberine was quantified by RP-HPLC and GC-MS method. Results showed a fair amount of berberine in all the plant samples. Higher berberine content was found in the root, followed by the stem, and least in leaves of the plant. Quantitative phytochemical analysis of parts of BLR plant samples revealed the presence of good amount of protein, phenol, flavonoid, total carbohydrates reducing, and non-reducing carbohydrate. Because of high amount of berberine (36mg/g dw) in BLR-5 root sample, it was further investigated for its biological activity both in-vivo and in-vitro. Moreover, to investigate the possible combinational therapy, Withania somnifera (WS) was cotreated with BLR hydroethanolic root extract. GC-MS analysis of BLR root extract showed 59% berberine, 8% ankorine and 5% canadine. In WS root extract oleic acid, phytol, hexadecanoic acid and squalene were found. A sub-acute study of 28 days was conducted against cyclophosphamideinduced immunosuppression and hepatotoxicity in rats. Cyclophosphamide was administrated orally in two doses of 100mg/kg/bw on the 9th and the 16th day of the study. Levamisole @50mg/kg/bw administrated subcutaneously for 28days, as a standard immuno-stimulatory drug. Oral administration of hydroethanolic root extract of BLR and WS with the doses of 200 and 400mg/kg body weight individually and in combination was given up to 28days in morning daily before meal. Rats were antigenically challenged with SRBCs for the investigation of immunological response. On the 14th and 28th day of study blood and tissue samples were examined for hematological, biochemical, immunological, and oxidative stress-related parameters. In-vivo study results showed that the plant extract treatment improved the hematological parameter, normalized liver function and kidney function test, and enhanced cellular and humoral immune response in rats in a dose-dependent manner on 28th day of study. Likewise, in-vitro antioxidant and anti-inflammatory studies showed low IC50 values for the combination of BLR and WS having higher inhibition of protein against heat denaturation. However, no antimicrobial activity of plant extract individually or in combination could be observed against tested animal and plant pathogenic microorganisms. Thus, it can be concluded from the present study results, that hydroethanolic root extract of BLR had ameliorating potentials against cyclophosphamide-induced toxicity and immunosuppression. However, co-administration of BLR and WS provided the advantage of producing maximum therapeutic efficacy as imunostimulatory, hepatoprotective, antioxidant and nephroprotective in rats through a synergistic mechanism of Berberis lycium Royle and Withania somnifera in cyclophosphamide-induced toxicity.
Open Access
In vitro establishment, biochemical profiling and cytotoxic evaluation in critically endangered herb Nardostachys jatamansi
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-02) Gupta, Palak; Kumar, Vandana A.
The medicinal herb Nardostachys jatamansi belonging to family Caprifoliaceae is a native of Himalayan region and grows at altitudes of 3300-5000 m asl. The growth of rhizome explant after 40 days of establishment in Woody Plant Medium supplemented with TDZ & GA3 (1 ppm & 0.5 ppm, respectively) showed 4.4 cm shoot length and 10.33 number of leaves. The roots& leaves collected in Oct-Nov (S-1), Jan-Feb (S-2) and June-July (S-3) from natural habitat and extracted in four different solvents viz. methanolic, ethylacetate, acetone and water were screened for phytochemical content and antioxidant activity. The roots of S-3 in methanolic extract showed the highest phytochemical contents like total phenol (63.76mg GAE g-1 extract), total tannin (204.67mg TAE g-1 extract) and ortho-dihydroxy phenol (18.8mg CE g-1 extract) whereas total flavonoid (48.5mg QE g-1 extract) was maximally reported in ethyl acetate. Highest antioxidant activities like total antioxidant (910mg AAE g-1), DPPH radical scavenging (IC50=57.26 μg ml-1), Superoxide radical scavenging (IC50=84.18μg ml-1), Hydrogen peroxide scavenging (IC50=74.34μg ml-1) and protein protective activity against free radical inducer AAPH were observed in S-3 methanolic root extract whereas FRAP (14.67 AAE g-1 extract) and Metal ion chelation (IC50=107.65μg ml-1) was best in S-3 acetone root extract. The highest number of compounds in S-3 root- (36) and leaf- (34) extracts were identified through GC-MS analyses with patchouli alcohol (22.92%) and α-Cadinol (14.17%) as predominant compounds. Highest content of Patchouli Alcohol (23.36 mg g-1 dry weight) was reported in S-3 root methanolic extract. Micronutrient analysis showed that Fe, Ca and Zn (19.95ppm, 10.30 ppm and 0.721 ppm) were highest in S-3 roots and Mn (4.27 ppm) in S-3 leaf was highest amongst the samples of different seasons. All the S-3 root extracts showed lowest MIC (15.62 & 31.25 μg) against gram positive L. monocytogenes and gram negative S. typhi, strains respectively. The maximum inhibition zone diameter (IZD) at par to ampicillin standard against 5 human pathogenic strains i.e., S. aureus (17.1 mm), B. cereus (16 mm), E. coli (8 mm), L. monocytogenes (14 mm) and S. typhi (10 mm) was recorded in S-3 ethyl acetate. The S-3 methanolic root extract showed maximum in vitro cytotoxic potential against three cancer cell lines viz. PK-15 (IC50=55.92μg ml-1), MDCK (IC50=64.55μg ml-1 ) and HEK (IC50=57.71μg ml-1). Results clearly revealed presence of significant amounts of phytochemicals and antioxidant activities due to variety of metabolites in leaves and roots and also in vitro cytotoxic potential against human pathogenic strains and cancer cell lines in roots. The multiutility of this medicinial herb against numerous pathological ailments was attributed to pharmacological activities.
Open Access
Studies on micropropagation with metabolite profiling for pharmacogenosic efficacy of Adina cordifolia and biochemical investigations of flowering in Dendrocalamus giganteus
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-02) Raypa, Pratima; Dubey, Ashutosh
Adina cordifolia a Rubiaceae family member is deciduous threatened tree and for in vitro propagation, axillary buds were augmented on MS medium supplemented with naphthalene acetic acid and 6- benzyl amino purine. Shoot length of propagated explants were found maximum on medium supplemented with 2 mg/L BAP or 0.5 mg/L NAA while maximum callus volume was induced in MS medium supplemented with 1.0 mg/L NAA, 1.0 mg/ L 2, 4- D and 0.5 mg/L BAP. On the metabolite profiling of Adina cordifolia different extracts through GC-MS, dominating constituents in the leaf extracts were trans squalene, vitamin E, phytol, and neophytadiene, while in bark extracts 7-hydroxycoumarin, nhexadecanoic acid, methyl commate, ergost-5-en-3.beta.-ol, -sitosterol. In root extracts major compounds, acetophenone, benzoic acid, 7-hydroxycoumarin, n-hexadecanoic acid, ergost-5-en-3.beta.-ol, beta.-sitosterol and gamma.-sitosterol were found. Through HPLC 7-hydroxycoumarin (7-HC), pharmaceutically important compound was found in high quantity in methonolic extracts of root, bark and calluses. Reducing power activity was positively correlated with all phytochemicals but best correlation was with phenols and flavonoids. FRAP activity were found positively correlated with phenolic content and flavonoid. Adina cordifolia extracts showed good cytotoxicity against human pathogenic strains i.e., Salmonella typhaemurium 52, Salmonella typhaemurium 149, Salmonella typhaemurium ATCC 51812, Salmonella entiritidis ATCC 13076, E. coli ATCC-25922 and E coli strain-13. Allelopathic effect of leaf extract of Adina cordifolia was studied on wheat genotypes as H2O2 content increment in wheat genotypes due to salt stress. Aqueous leaf extract of Adina cordifolia treatments decreased H2O2 content with increase in proline content. Flowering gene expression analysis in bamboo genotype Dendrocalamus giganteus was carried out by RT-PCR analysis. The transcript accumulation of homolog of CRABS CLAW, SEPALLATA1, AGAMOUS and PISTILLATA in flowering bud of Dendrocalamus giganteus was 3.6, 1.41, 3.02 and 5.46 folds higher respectively as compared to non-flowering bamboo leaves of Dendrocalamus giganteus. Metabolite profiling of D. giganteus revealed the presence of dominating constituents such as -thujene, vitamin E, 6-methyl-2-(5-methyl-5-vinyltetrahydro-2-furanyl)-5-hepten-3-one, elemol, -eudesmol, 7-epi- -eudesmol, n-hexadecanoic acid, n-heneicosane, n-tetracontane, campesterol, -sitosterol. Thus these results on A. cordifolia would be used to set new standard of prescribing the dosage of herbal drugs to propose an alternative to modern system of medicines and bamboo flowering gene expression analysis provide good understanding of flowering gene expression pattern.
Open Access
Investigation on biochemical basis of resistant starch with respect to physiochemical properties in cereals-millets, rice and wheat
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-12) Verma, Vivek Chandra; Agrawal, Sanjeev
The present investigation was carried out to assess the physio-chemical properties of starch attributing towards resistant starch in millets (Barnyard millet; VL Madira-207, Finger millet; VL Mandua-352 and Foxtail millet; DHFT-109-3), rice (PB-2 and PD-19) and wheat (PBW-343 and UP-262). The study emphasize on the factors which influences the resistant starch in cereals. In the present study starch isolated from different cereals were viewed under SEM to analyse the shape and size of starch granules. The size of starch granules were diverse range in cereals like millets (4.6-12.23 μm) rice (6.22-177.13 μm) and wheat (2.50-198.23 μm). Small and large spherical, irregular and polygonal shape of starch granules was observed. The FTIR bands at 1047 and 1022 cm-1 gives ordered and amorphous structure of starches and absorbance ratio at 1047/1022 gives indexes of the short-range order of double helices ranged from 0.94-1.24. Total starch (57-70%), amylase (31-45%) and amylose-amylopectin ratio (0.46-0.83) was determined in different cereals under observation. The amylose content was positively correlated with resistant starch at P < 0.01 while total starch had non-significant impact on resistant starch. The other nutrient of cereals like dietary fibre (2.66-6.98%), total soluble protein (9.33-16.29%) and soluble protein fraction like albumin (1.41-2.44%), globulins (1.25-2.23%) and prolamines (2.04-2.88%) was estimated to establish the correlation with resistant starch. These other nutrient of cereals showed positive correlation with resistant starch at P < .01. In vitro digestion of native starch, gelatinized starch and retrograded starch was done at time interval of 30, 60, 90, 120 min and it was found that the rate of enzymatic digestion follows the order; Gelatinized starch > Native starch > Retrograded starch. During endosperm developmental stages at S1, S2, S3, S4 (7, 14, 21, 28 DAA) the key enzymes for starch biosynthesis like AGPase, SSS and GBSS were estimated. The activity of starch biosynthetic enzymes follows the order AGPase > GBSS > SSS in all these varieties during endosperm development. The higher activity of AGPase, GBSS and SSS found in rice comprises higher accumulation of amylose and total starch during endosperm development. The accumulation of amylose and total starch in millets was found at par to the wheat varieties which is related to the activities pattern of enzymes. All three enzymes under study showed the positive correlation with each other during endosperm development. Accumulation of amylose showed positive correlation with AGPase and GBSS, while non-significant correlation with SSS. Total starch accumulation showed positive correlation with AGPase.
Open Access
Optimization of pretreatment parameters and nanoparticle based enzyme immobilization for lignocellulosic biomass saccharification
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-09) Singh, Archana; Verma, A.K.
Pretreatment is a prerequisite technology to facilitate the recovery of fermentable sugars from lignocellulosic biomass. Various lignocellulosic biomass viz. sugarcane leaves, rice straw, wheat straw, orange peel and pomegranate peel were used for pretreatment with ionic liquid EmimAc (1-ethyl-3-methylimidazolium acetate), aqueous ammonia and a combination of both followed by saccharification with immobilized cellulase and β-glucosidase (BGL) on magnetic nanoparticles. Magnetic nanoparticles (Fe3O4) and cellulase/BGL immobilized nanoparticles were characterized by TEM, SEM, FTIR and Zeta potential. Immobilization efficiency of cellulase and BGL were found to be 86.36% and 87.73%, immobilization yield were 83.38 and 85.18% respectively. The immobilized cellulase had optimum pH 5.0 and optimum temperature 50°C. The immobilized cellulase retained 90% of initial activity after 60 min incubation at 50°C while free cellulase had only 49% of its initial activity. Km value for free and immobilized cellulase was found to be 0.625 mM and 0.714 mM and the Vmax was 35.71 and 38.16 μmole/min/mg respectively. Immobilized cellulase retained up to 97% of residual activity on 6th day when stored at 4°C and 45% at 25°C whereas free cellulase had 86% of residual activity at 4°C and 20% at 25°C. Immobilized cellulase retained about 87% of residual activity after 10 cycle of reuse. The immobilized BGL had optimum pH 5.0 and optimum temperature 50°C. The immobilized BGL retained 91% of initial activity after 60 min incubation at 50°C while free BGL had only 51% of its initial activity. Km value for free and immobilized BGL was found to be 0.555 mM and 0.779 mM and the Vmax was 38.46 and 43.47 μmole/min/mg. Immobilized BGL retained up to 95% of residual activity on 6th day when stored at 4°C and 42% at 25°C whereas free BGL had 87% of residual activity at 4°C and 18% at 25°C. Immobilized BGL retained about 85% of residual activity after 10 cycle of reuse. The combined use of EmimAc and aqueous ammonia in 1:1 for pretreatment at 6 hrs with <2 mm size of biomass followed by saccharification with 3.2 U of immobilized cellulase showed a synergistic effect on biomass with release of 93% glucose. Combined pretreated biomass were analysed for saccharification by addition of 3.2 U BGL with 3.2 U cellulase and 97% of glucose released. Furthermore, EmimAc and immobilized cellulase were recycled for 6 times with 89% of efficiency; EmimAc, immobilized cellulase and immobilized BGL were recycled for 6 times with 90% of efficiency. Thus, compared to the other pretreatment method, this combine method is more efficient and cost effective. To identify the cheapest source for enzymes biogas slurry was analysed for different enzymes. These were cellulase (0.41), BGL (0.30), Xylanase (0.31), α-amylase (0.06), β-amylase (0.20), Dehydrogenase (0.37), and Protease (0.29) U/mL. Saccharification was done with crude enzyme isolated from biogas slurry and found 69% of glucose release in combined pretreated biomass.
Open Access
Effect of micro/macro environment of rhizogenesis for optimization of withanolide contents in Withania somnifera
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Singh, Manali; Agrawal, Sanjeev
India is a varietal emporium of medicinal plants and is one of the richest countries in the world with regard to genetic resources of medicinal plants. Withania somnifera is a revered herb of Indian medicinal system. Owing to the presence of pharmacologically bioactive compounds. The roots and leaves of Ashwagandha contain various alkaloids viz. Withanolides and withaferin A. Among withanolides, withaferin A and withanolide A has been reported to be dominant metabolite distributed among various tissues of this plant in varying concentrations. Since the location and environmental factors play a key role in the biosynthesis of the bioactive metabolites so the agro-climatic conditions are conductive for introducing and domesticating new exotic plant varieties with enhanced and consistent withanolide A/withaferin A contents. Thus keeping in mind the pharmacological significance of the bioactive ingredients, the present study focuses on the different strategies employed for their enhanced production. MS medium supplemented with Kinetin 1.5 mg/l was found to have highest withaferin A content i.e. 0.93 mg/g (DW) while withanolide A content was highest in MS medium supplemented with Kinetin 1.5 mg/l i.e 0.348 mg/l (DW). Agrobacterium rhizogenes transformed Poshita recorded highest total withaferin A content i.e 5.969 mg/g (DW) and total withanolide content i.e 2.017 mg/g(DW). In aquaponics the total withaferin A content of 6 month grown seedling, was higher in Jawahar-20 i.e 2.904 mg/g (DW) while withanolide content was higher in Poshita i.e 4.019 mg/g (DW). Moreover use of different elicitors at very low concentrations were found to enhance the withaferin A /withanolide A content. Thus both the varieties i.e. Jawahar-20 and Poshita are the promising varieties of Withania somnifera, with higher withaferin A /withanolide A production. Thus implementation of tissue culture techniques, Hairy root culture, usage of elicitors, bio augmented soil and aquaponic system can offer promising approach for enhanced growth and sustainable production of pharmacologically bioactive ingredients in perspective of improving cultivation of Ashwagandha.
Open Access
Characterization and comparison of bioactive constituents in the in vitro regenerated and wild roots of Himalayan medicinal herb Valeriana jatamansi Jones
(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2018-08) Saini, Soniya; Kumar, Vandana A.
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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