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2021 - 20224
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Subject: Biochemistry × End date: 2024 × Start date: 2020 × Type: Thesis × Thesis Type: Ph.D ×
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    ThesisItemOpen 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.
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    ThesisItemOpen 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.
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    ThesisItemOpen 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.
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    ThesisItemOpen 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.