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ThesisItem 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, HimanshuBrassica 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.ThesisItem Open Access Evaluation of pyrolyzed pine needle bio-oil for antifungal and nematicidal activities with molecular docking analysis on target proteins(G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-09) Chandra, Kailash; Verma, A. K.Pyrolyzed pine needle bio-oil formulation was used to evaluate antifungal and nematicidal activity. Antifungal activity of pine needle bio-oil (40%) based formulation and its four dilutions (10-1, 10-2, 10-3 and 10-4) were evaluated in both condition in-vitro and in-vivo. In-vitro activity of the formulation and its 10-1 showed 100% inhibitory activity against Alternaria brassicae while in 10-2, 10-3 and 10-4 dilutions the inhibitory activity was 52%, 48% and 42% respectively. The pre-conditioning brassica plants treated with the foliar spray of antifungal pine needle bio-oil formulation showed 100% inhibition of Alternaria brassicae while in post-conditioning plants treated with dilutions 10-1, 10-2, 10-3 and 10-4 were showed disease severity of about 3.3%, 6.6%, 13.3% and 30% respectively. Biochemical analysis of pine needle bio-oil formulation treated Brassica leaves revealed that the chlorophyll and carbohydrates content of post-conditioning plants increased with respect to the increase in bio-oil dilutions while protein content was found to be decreased. In case of post-conditioning plants, only protein content was increased while the chlorophyll and carbohydrates content was found to be similar. Molecular docking analysis of four compounds present in pine needle bio-oil, 4-methyl-pentanoic acid and nonanoic acid with succinate dehydrogenase while 1-(1H-Imidazol-4-YL)-Ethanone and dehydroabietic acid with lanosterol 14α demethylase of Alternaria brassicae. Showed dock score -4.03 and -6.9 Kcal/mol for the enzyme succinate dehydrogenase and -9.6 and -6.8 Kcal/mol for lanosterol 14α demethylase respectively. Nematicidal activity of pine needle bio-oil based formulation was analyzed against root knot nematode Meloidogyne spp. The concentration of formulation 5%, 7.5% and 10% showed maximum 100% inhibition against mortality, immobility and egg hatching of Meloidogyne spp. Molecular docking analysis of four compounds present in pine needle bio-oil, dehydroabietic acid and 2-methyl phenol with thymidylate synthase while dehydroabietic acid and 4-Methyl Pentanoic acid with Na-K ATPase of Meloidogyne spp. Showed dock score -7.1 and -4.94 Kcal/mol for thymidylate synthase and -5.0 and -4.8 Kcal/mol for Na-K ATPase respectively. Hence the pine needle bio-oil based formulation is more effective against Alternaria brassicae nematode Meloidogyne sppThesisItem Open Access Biochemical appraisal of differentially treated Banana (Musa acuminata L.) cv. Grand Naine for post-harvest quality(G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-09) Mathur, Ananya; Vandana A. KumarThe study conducted during 2022, in biochemistry department, cbsh, gbpuat, was aimed to enhance the shelf-life of banana (musa acuminata l.) cv. grand naine by applying aqueous solutions of salicylic acid (s: 1mm), gibberellic acid (g: 1mm), 1- methylcyclopropene (m1 to m5, ranging between 50μl/l to 800 μl/l) and salicylic- +gibberellic acids (sg1: 1mm each and sg2: 1mm of salicylic- and 2mm gibberellic acids) for 15 minutes followed by air drying and storage for 20 days at room temperature (24ºc- 27ºc). enzymatic studies, biochemical parameters, sensory evaluation and physical observations, in triplicates were performed at 5day-intervals. the increase in activities of cellulase, invertase, xylanase, polygalacturonase and peroxidase (0.071±0.006, 0.002±0.001, 0.0206±0.013, 0.004±0.0001, 0.030±0.001 u/100gfw, respectively, on day 20) viz-a-viz control (0.365±0.002, 0.0041±0.01, 1.7462±0.016, 0.014±0.001, 0.0520±0.001, respectively, on day 20) and contents of total soluble protein, soluble sugars and total phenols (0.308g/100g, 3.23%, 61.2mgae/100g, respectively, on day 20) was the least for m5 treatment and highest for s treatment. all mcp treatments delayed ripening as they performed better than s, g, sg1 and sg2, throughout storage. the decrease in total starch, resistant starch, total flavonoids, titratable acidity and ascorbic acid (72.73%, 53.9%, 15.5mqe/100g, 0.7%, 85±0.04mg/100ml, respectively, on day 20) was the least for m5 treatment. other treatments of mcp also decreased total starch, resistant starch and total flavonoid contents, concomitant to their increasing concentrations. the decrease in activities of cellulase, xylanase, polygalacturonase, soluble sugars and increase in contents of total starch, resistant starch and total flavonoids by sg1 & sg2 were significantly lower than those shown by s & g alone. delayed color and texture changes were observed in m5 treated banana which remained ‘hard’ (texture score: 1) and ‘yellow with green tips’ (color score: 2) till 10 days of storage. an enhanced shelf-life of banana was observed for 10-15 days when treated with 800 μl/l of mcp treatment and stored at room temperature.ThesisItem Open Access Biochemical evaluation of ethnomedicinal tree Oroxylum indicum (L.) Kurz(G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-09) Pant, Samridhi; Dubey, AshutoshOroxylum indicum belongs to the monotypic genus Oroxylum. It has long been used in traditional medicinal systems and is an important part of Ayurveda. In the present research the antioxidant activity and the secondary metabolites’ constitution of seed and stem bark of O. indicum in the aqueous and hexane extracts was studied. Total antioxidants (12.792±0.1 mgAAE/g, 14.961±0.04 mgAAE/g), total phenolic (136.181±0.2 mgGAE/g, 135.08±0.15 mgGAE/g), total tannin (56.999±0.8 mgTAE/g, 26.833±0.4 mgTAE/g), total flavonoid (88.911±0.41 mgQE/g, 122.925±0.30 mgQE/g) and total flavonol content (25.832±0.23 mgCE/g, 27.885±0.087 mgCE/g) was higher in aqueous extract for both seed and stem bark than hexane extract, respectively. Total antioxidants showed positive correlation with the phytochemicals. The metal ion chelating activity (IC50) (32.404±0.14 μg/mL and 30.756 ±0.27 μg/mL), DPPH scavenging (IC50) (101.396±0.8 μg/mL and 115.358±0.4 μg/mL) and reducing power (RP50) (37.115±0.02 μg/mL and 40.134±0.5 μg/mL) was also higher in the aqueous extract of seed and stem bark than the hexane extracts as expressed by their lower IC50 values respectively. All activities showed a negative correlation with the phytochemicals indicating the increase in the phytochemicals will lead to better antioxidant potential by lowering the IC50. The extracts were tested for their anti-inflammatory activity. The results were expressed as IC50 values which were 82.26±0.5 μg/mL and 79.681±0.4 μg/mL for the aqueous extract which was better than the hexane extract. Anti-bacterial activity of the extracts was tested against Xanthomonas oryzae and activity was observed only in the aqueous extract of seeds. A maximum zone of inhibition of 18 mm was observed at 5000 μg/mL. The seed proteins of O. indicum were fractionated and tested for their amylase and protease activity and inhibition. Highest amylase activity was observed in albumins and highest inhibition was observed in the prolamin. For the protease activity, albumins and globulins showed almost same protease activity but the highest inhibition was observed in globulins. The seed proteins of O. indicum seeds were partially purified using ammonium sulphate precipitation and dialysis and characterized using SDS-PAGE. The process showed 9 bands on the gel with molecular weight ranging from 97 kDa to 14.3 kDa.ThesisItem Open Access Studies on clonal diversity in eucalyptus using biochemical and molecular approaches(G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-09) Belamana, Bhargavi; Dubey, AshutoshEucalyptus is a fast growing evergreen tree with its rapid growth and capacity to satisfy rural needs for fuel and shelter, its plantations were widely encouraged. The current work details about the evaluation of the clonal diversity through biochemical and molecular parameters among five Eucalyptus clones acquired from Agro Forestry Research Centre, GBPUAT, Pantnagar. Total phenol content was observed to be lowest (109.6±0.05 mg GAE/g) in sample PES-21-4 and highest (132±0.09 mg GAE/g) in sample K 25. Total antioxidant capacity was observed to be lowest (15.22±0.006 mg/g) in sample PES-21-3 and highest (17.89±0.02 mg/g) in sample PES-21-2. Total flavonoid content was observed to be lowest (19.48±0.2 mg CE/g) in sample PES-21-3 and highest (24.81±0.3 mg CE/g) in sample PES-21-1. Total tannin content was observed to be lowest (9.12±0.05 TAE/g) in sample PES- 21-1 and highest (10.17±0.13 TAE/g) in sample PES-21-4. IC50 value of DPPH activity was observed to be lowest (23.49±0.009 μg/ml) in sample PES-21-3 and highest (26.01±0.005 μg/ml) in sample K25. IC50 value of metal ion chelating activity was observed to be lowest (17.76±0.11 μg/ml) in sample PES-21-3 and highest (19.86±0.03 μg/ml) in sample K25. Amylase inhibition was found to be highest in hexane extract (46.04±0.11%) than the methanolic extract (34.59±0.03 %). SDS-PAGE analysis revealed 15 bands out of which the most prominent band obtained was band 14 of size 92.7kDa with Rf value of 0.277. Five Eucalyptus clones were characterized at molecular level using eight RAPD primers. The minimum similarity coefficient was between clone PES-21-1 and clone PES-21-4 (0.63), while maximum similarity coefficient was between clone PES-21-3 and clone PES-21-4 (0.79). Dendogram revealed that Clone PES-21-3 and clone PES-21-4 are showing high similarity while Clone K 25 was quite diverse from rest of the clones. The quantitative determination of the phytochemical composition and their activities in Eucalyptus helps to understand its therapeutic properties and utilizing molecular markers to determine the clonal diversity leads to a better knowledge about plant evolutionary history and provide valuable information about agrobiodiversity dynamics.ThesisItem Open Access Investigation on phytochemical attributes and antioxidative potential of Pleurotus sajor-caju (Fr.) Singer(G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-09) Srinivas, Arji Ravi Chandra; Punetha, HimanshuThe current work details about the evaluation of the phytochemical composition of different polarity extracts and the biological activities of mushroom P.sajor-caju sample acquired from mushroom research and training centre, G. B. Pant University of Agriculture & Technology, Pantnagar. Twenty one compounds were identified with the help of GC-MS representing 98.61% of total methanolic extract composition. Cis-Vaccenic acid, Beta-Monolinolein, ergosterol derivatives are major compounds identified. Fifty two compounds were identified with the help of GC-MS representing 99.48 % of total petroleum ether extract composition. Linoleic acid, Eicosane, Heneicosane and Hexatriacontane were the major compounds identified. The Methanolic extract showed potent antioxidant activity in terms of DPPH radical scavenging, metal chelating activity and reducing power activity having IC50 value of 677.73±1.72μg/mL, 4.25±0.05 mg/mL and 9.66±0.11 mg/mL respectively in comparison to petroleum ether extract which might be due to presence of Cis-Vaccenic acid, Beta-monolinolein. Total phenolic content (TPC), Total flavonoid content (TFC) and Total antioxidant content (TAC) of methanol and petroleum ether extract were also evaluated. TPC, TFC and TAC were higher in methanol extract having 21.3±0.006 mg/gm, 3.43±0.002 mg/gm, and 19.8±0.65 mg/gm, respectively. Crude protein content was observed as 19.78% along with mineral composition Fe, Mn, Cu and Zn as 0.0156±0.89 mg/g, 0.029±1.56 mg/g, 0.018±1.25 mg/g and 0.085±1.23 mg/g respectively. The molecular docking of bioactive compounds Ergosterol and β-glucans of P sajor-caju against Covid-19 spike proteins 6vsb, 6lxt, 6lvn and 6vw1 was performed using PyRx software by autodock wizard as the engine for docking. The results obtained shows binding energies of 6vsb, 6lxt, 6lvn and 6vw1 with ergosterol as -9.3 kcal/mol, -7.9 kcal/mol, -7.1 kcal/mol and -8.7 kcal/mol respectively. The binding energies of 6vsb, 6lxt, 6lvn and 6vw1 with β-glucans as -6.8 kcal/mol, -7.5 kcal/mol, -6.2 kcal/mol and -7.5 kcal/mol respectively. The mushroom P. sajor-caju thus have potent medicinal properties and if explored further might be potent antioxidant and antiviral cost-effective agent compared to synthetically derived agents from pharma industries.ThesisItem Open Access Effect of gamma Iiradiation on quality of wheat and buckwheat flour(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2022-09) Mehra, Dakshita; Agrawal, SanjeevCereals and pseudocereals constitute a large part of human diet. The quality of flour during storage gets deteriorated by change in the temperature conditions, moisture, fungus development, pest infestations and many other variables. Gamma rays have been used to avoid food deterioration. The present study was undertaken to evaluate the quality of flour after exposing it at different dose of gamma radiation. The effect of gamma irradiation doses (1, 2, 5, 7 and 10 kGy) on quality of wheat and buckwheat flour was studied. The nutritional, antioxidant and antinutritional parameters like crude protein content, total starch content, amylose, amylopectin content, amylose to amylopectin ratio, resistant starch, total phenol, flavonoid, total antioxidant content, tannin and phytic acid content in non irradiated wheat varieties ranged from 11.15%-13.14% , 61.39-67.08 gm/100g, 17.62-22.62 gm/100g, 82.38-77.38 gm/100g, 0.21-0.29, 0.50-0.74 gm/100g , 1.19-1.38mg GAE/g, 0.22-0.36 mg QE/g, 12.44-16.22 μmole of vit C/gm, 8.48-6.03 mg TAE/g and 280.48-253.65 mg/100g, respectively whereas in buckwheat it was found to be 11.22%, 71.41 gm/100g, 24.62 gm/100g, 75.38 gm/100g, 0.32, 0.85 gm/100g, 4.41 mg GAE/g, 3.92 mg QE/g, 19.65 μmole of vit C/gm, 14.84 mg TAE/g and 235.47 mg/100g, respectively. After exposure to gamma radiations, no significant difference was observed in crude protein and total starch content, whereas amylose, resistant starch, phenol, flavonoid and total antioxidant content were increased significantly (p≤0.05) in both wheat and buckwheat with increasing doses. Tannin, phytic acid and amylopectin content were found to decrease on exposure of irradiation. A positive high correlation was observed between total phenol, flavonoid and antioxidant content in both irradiated wheat and buckwheat. It may be inferred that gamma irradiation doses does not deteriorate the quality of wheat or buckwheat flour however, it may improve the quality by decreasing the antinutritional contents and increasing the antioxidants.ThesisItem Open Access Biological activities of extracts and essential oils of Anaphalis margaritacea- (L.) Benth. & Hook.f. in relation to its phytochemical composition from hilly terrains of Uttarakhand Himalaya(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2022-09) Arti; Dubey, S.K.Present research details the phytochemical composition of essential oils, as well as biological activity of the plant Anaphalis margaritacea in different polarity extract, collected fromKumaun region of Uttarakhand at an elevation of 2,200m. Total 67compounds were revealed in essential oil via GC-MS, however 8 remained unidentified, comprising 94.04% of the total essential oil composition. The major compounds in the essential oil comprised of Caryophyllene (E) (17.82%), Asarone (8.89%), humulene (8.27%), and caryophyllene oxide (7.00%). Thirty four compounds were identified constituting 89.09% of the total methanolic extract which include 5R, 8R, 9S, 10R)-2-Formyl-3-Hydroxy-5-Isopropeny-8-8Methyl (3a10)- octahydronaptho (13.15%), oleic acid (11.31%) etc, Identified compounds represented 70.21% of the total hexane extract composition with prominent compounds Hexatriacontane (20.43%), Tetracontane (14.70%), Heneicosane (2.66%), and Tetratetracontane (2.05%). Total phenolic content (TPC), Total flavonoid content (TFC) and Total antioxidant content (TAC) of methanol and hexane extract were also evaluated. TPC, TFC and TAC were higher in methanol extract having 19.11±0.24, 34.07±1.54, and 28.09±1.82 (mg/gm), respectively. DPPH radical scavenging was expressed as IC50 values which were 75.94 ±0.66 μg/mL in methanolic extract, 141.42 ± 0.73μg/mL in hexane extract and 57.66 ± 0.40 μg/mL in standard ascorbic acid. For metal chelating activity, the IC50 value in methanolic extract, hexane extract and standard EDTA was 72.90±0.096, 141.42±1.25, 60.18± 0.39 (μg/mL) respectively. For reducing power activity, the RP50 value of gallic acid, methanolic and hexane extract were 74.37±0.29 μg/mL, 79.64±5.36 μg/mL, 107.0682±1.34 μg/mL respectively. Methanolic and hexane extracts exhibited potent anti-inflammatory activity with IC50 values of 37.58±1.05 μg/mL, 46.92±0.67 μg/ mL respectively. Diclofenac sodium salt used as standard (IC50 value 51.59±1.18 μg/mL). The results suggest that A. margaritacea have potent biological activities and can be a powerful antioxidant and anti-inflammatory agent if explored further. The plant extracts might be more affordable than synthetically generated compounds, eco-friendly, and economically viable for the pharmaceutical and biomedical industries.ThesisItem Open Access Biological activities of essential oil and whole plant extracts of Callicarpa macrophylla Vahl. in relation to its phytochemical composition(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2022-08) Arya, Aradhana; Arya, Aradhana; Dubey, S.K.; Dubey, S.K.The present study gives a detailed explanation of the phytochemical profile of essential oil and different polarity extracts along with the evaluation of the biological activities of the plant Callicarpa macrophylla collected from the hilly regions of Uttarakhand from the elevation of 1394 m. Fourty-five compounds were identified by GC-MS accounting for 78.79% of the total essential oil composition in which β- Selinene (15.24%) was the prominent constituent. The methanolic extract of the whole plant comprised of 26 compounds constituting 79.74% of the total methanolic extract composition. The major compounds in methanolic extract were Trachylobane (31.87%) and Palmitoleic acid (8.26%). Thirty-two compounds were identified in the hexane extract comprising 63.56% of the total composition. The prominent compounds in hexane extract were Tetratetracontane (9.78%) and Pentatriacontane (7.15%). The Total phenolic content (TPC), Total flavonoid content (TFC) and Total antioxidant content (TAC) was estimated in higher quantities in methanolic extract having 51.27±0.61 mg/gm, 49.47±0.34 mg/gm and 27.89±0.24 mg/gm, respectively; as compared to that of hexane extract having 40.58±0.50 mg/gm, 34.42±0.27 mg/gm and 11.42±0.23 mg/gm, respectively. Methanolic extract demonstrated potent antioxidant activity in terms of DPPH radical scavenging and reducing power activity having the IC50 and RP50 values of 91.97±0.33 μg/mL and 56.72±0.67 μg/mL compared to standards Ascorbic acid (57.66±0.40 μg/mL) and Gallic acid (74.36±0.29 μg/mL) respectively. Both methanolic and hexane extract showed better metal chelating activity; with IC50 value of 10.90±1.40 μg/mL and 41.60±1.31 μg/mL, respectively as compared to the standard EDTA 60.18±0.39 μg/mL. The methanolic extract displayed potent anti-inflammatory activity with an IB50 value of 64.09±1.31 μg/mL comparable to that of the standard Diclofenac Sodium (51.69±0.99 μg/mL). Callicarpa macrophylla possesses a strong therapeutic potential and, if explored further, may prove to be a powerful antioxidant, anti-inflammatory, and cost-effective agent compared to synthetically derived and can replace various artificial synthetic agents from pharmaceutical industries.
