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ThesisItem Open Access Screening of tea cultivars grown in Kumaon region for their total phenolics, pigments and individual catechin contents(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2005-08) Pandey, Sudhanshu; Mishra, D.P.Total chlorophylls, chlorophyll a , chlorophyll b, total carotenoids and total phenolics contents and HPLC profiling of various catechins and quantitative estimation of (-)EGCG,(-)EC and (-)ECG were carried out in fresh tea leaf leaves (dried immediately after sampling), of 6 varieties grown in Kumaon region and a black orthodox tea sample processed locally. The main objective of the study was to screen these varieties for their catechin contents and other flavanoid constituents. Dried leaf powder was used to estimate chlorophylls, carotenoids and total phenols. For catechins, dried leaf powder was first extracted with aqueous methanol. This extract was subjected to separation of catechins on HPLC and identification and quantitative estimation were done using standard catechins. Contents of various parameters were variable among varieties and there was no set pattern. Kangra Jat had highest content of chlorophyll a, and total carotenoids while chlorophyll b, total chlorophyll and total phenols were highest in UPAS1-9. T-78 exhibited lowest content of all these parameters. Black orthodox tea had pretty high content of chlorophylls and carotenoids but its phenolics content was lowest, perhaps due to oxidation of phenolics during processing. The HPLC profiles gave eight peaks, out of which first two peak, were probably of some non-catechin compounds, hence ignored. Out of remaining 6 peaks, 3 were of (-)EGCG,(-)EC and (-)ECG. Other three were named as U1, U2 and U3.Out of them U1 was supported to be (-)EGC. By comparison of peak areas,retention times etc., quantitative estimation of (-)EGCG, (-)EC and (-)EC was performed, which revealed these three catechins comprised nearly 0.11-0.14% of dried solids (110-140 mg g-1) and (-)EGCG was nearly (69-82 mg g-1),half of all catechins estimated. On the basis of increasing catechin contents, order of varieties was: Black Orthodox tea(commercial)ThesisItem Open Access Biochemical evaluation of genotypes of Jatropha curcas L., and optimization of micropropagation schedule(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2005-07) Munagapati, Sandhya; Gaur, A.K.Five genotypes of Jatropha curcas viz. shu03001, shu03002, shu03004, shu04001 and shu04004 were selected for their further identification and biochemical characterization. Identification of genotypes had been done on the basis of banding pattern on SDS-PAGE of total seed storage protein and its fractions viz. globulins and albumins. Based on the Jaccard‟s similarity coefficient obtained from the banding patterns of SDS-PAGE, the five genotypes are grouped into two categories. Group1 consisting of shu03001, shu03004 and shu04004 and Group2 consisting of shu03002 and shu04001. These results were based in accordance with the percentage protein content of the respective genotypes taken for study and the oil contents of these genotypes were almost comparable. However, the lipid/protein ratio was highest in shu04001. In view of optimizing micropropagation schedule, the experiments were also conducted on three genotypes of J.curcas viz. shu03001, shu03002 and shu03004 to study the effect of various parameters viz. genotypes, growth regulators and explants and their interactions on callus induction frequency. Two explants viz., leaf and petiole from all the three genotypes were placed on different combinations of growth regulators. Calluses were induced from the two explants on all types of callus induction media containing four levels of 2,4-D (1, 2, 2.5and 3mg/l) with BAP (0.1,0.2,0.25 and 0.3mg/l) and two levels of NAA (1 and 2 mg/l) with two levels of BAP (0.1 and 0.2 mg/l). Maximum callus induction frequency of 92.8, 93.1; 97.4, 98.7 and 88.8, 96.1% was observed for leaf and petiole explants of shu03001, shu03002 and shu03004 genotypes respectively. In a generalized way, it was observed that shu03004 genotype showed highest callus induction frequency followed by shu03002 and shu03001. Media with NAA (1mg/l) and BAP (0.1mg/l) showed highest callus induction frequency with both leaf and petiole explants in all the three genotypes. Amongst explants, petiole showed higher callus induction frequency followed by leaf explant on all types of callus induction media.ThesisItem 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, AshutoshAdina 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.ThesisItem Open Access Eco-technological potential assessment of renewable bamboo biomass for bio-energy and sustainable environmental conservation(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-08) Ray, Avedananda; Dubey, AshutoshThe present work focused on the energy coversion efficacy of leaves of bamboo sp. Viz., Bambusa balcooa, Dendrocalamus hamiltonii, Dendrocalamus asper. After Pyrolysis of biomass 1. Bambusa balcooa bio-oil was further analysed for fuel property and bio-physical characterization, 2. biochars from Bambusa balcooa, Dendrocalamus Hamiltonii were tested for heavy metal remedation and other soil amendment experimentation and 3. biochar and bio-tar from Dendrocalamus asper were used for energy efficient pallet formation. The chemical composition of pyrolytic oil consisted of phenolic compounds contributed towards acidity of bio –oil and its relative density was found 0.912, Kinematic viscosity of 4.72, Cloud and pour points were -9 and -18 0C respectively, Flash and fire points were 59.2 and 68.5 0C respectively, ash content was 0.11, Conradson Carbon residue was 9.6 and acid value was 13.5. Thermal kinetics modeling of Bambusa balcooa biomass drying using TGA showd that Henderson model was suitably fitted (Highest R2 values) and actiovation energy for initial drying was calculated around 64.45 kJ/mol.For the devolatization stage, activation energy was found 40.36 kJ/mol.Physical study (FTIR, SEM, BET) study showed that biochar from bamboo could be used for adsorption of heavy metals. Dendrocalamus hamiiltonii biochar had equilibrium time at 60 min for Co (II) ions and 30 min for Pb (II) ions, dosage of 0.6 g/100 m.l for Co and 0.5 g/100m.l for Pb, pH 7 for max-imum uptake of Co(II) ions and pH 5 for maximum uptake of Pb (II) found optimum. Second order kinetic model, Freundlich isotherm model was found suitable to adsorption process. Response Surface Methodology applied for the adsorption from zinc electrode wastewater by Bambusa balcooa biochar showed that the optimum adsorption conditions pH (A) = 7, Time (B) = 73.05 , Biochar Dose (C) = 0.98) with predicted adsorption 18.32% (Pb), 78.39% (Zn), 44.5% (Cd), 95.71%(Cr). Calorific value of pellet produced from biochar and biotar was in range of 20-30 MJ/Kg. Ammonia adsorption was higher (123.08%) in case of 1: 1 soil : biochar ratio. Bambusa balcooa biochar applied soil has 34% increment in water holding capacity than the untreated soil.Ethanol production from pretreated biomass of Bambusa balcooa leaves (Cellulose rich fractions) was done through biomass saccharification through Cellulases and Saccharomyces cerevisiae inoculation with an overall raw biomass to ethanol conversion efficiency was 36.72%.ThesisItem Open Access Screening and biological activities of various bioactive compounds in finger millet [Eleusine coracana (L.) Gaertn.] genotypes collected from various hilly regions of Uttarakhand(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-07) Bhatt, Dheeraj; Verma, K.C.Many food crops with high quality nutritional value growing in the Himalayan region have been in use from ancient time to treat various ailments in different traditional ways. In recent year, demand for high nutritional value crops has increased tremendously. The present study, focused on the Screening and Biological Activities of Various Bioactive Compounds of 10 different genotypes of Finger millet [Eleusine coracana (L.) Gaertn.], commonly known as Ragi, collected from different regions of Uttarakhand. The proximate composition showed variation in value for biochemical constituents like ash (2.58-3.18%), moisture (6.41-11.12%), protein (6.20-11.12%), fat (2.58-4.56%), tannin (1.12-4.71%), starch (35.71-41.57%), resistant starch (5.85-12.71%) and minerals (mg/100g) like as Ca (96.73-230.78), Mg (122.2-221.32), Mn (152.61-369.14) & Fe (0.147-9.44). The % yield in methanol extracts were range from 3.08-6.12% while 2.81-5.75% in hexane extracts. The range of total phenolic, total flavonoid, total antioxidant content, DPPH free radical scavenging activity, metal chelating activity of Fe2+, ferrous reducing antioxidant power , in-vitro antiinflammatory activity in methanol were 3.17-5.01 mg GAE/g, 0.76-1.72 mg CE/g, 39.02-85.98 mM AA/mg, 43.37-48.84%, 20.73-35.78%, 30.91-109.38 μg BHTE/mg and 38.98-73.75% respectively whereas in hexane, the range were 1.76-3.16 mg GAE/g, 0.511-1.29 mg CE/g, 30.31-72.41 mM AA/mg, 27.48-42.8%, 17.34-31.47%, 16.21-83.72 μg BHTE/mg and 21.99-64.77% respectively. The seed extract analysis by GC-MS reveled active components that show all sort of pharmacological activities from antioxidants to anti-cancerous. Out of all genotypes, the genotype which was collected from Bageshwer, shows better activity in terms of biochemical compositions like fat, tannin, starch & resistant starch contents as well as in antioxidant potential like TPC, TFC, TAA and ferrous chelating activity followed by Dwarahat genotype which was good in term of mineral composition (Ca, Mg & Fe) and also shows better ferrous reducing antioxidant power and in-vitro anti-inflammatory activity. As finger millet has significant mineral composition and shows good antioxidant and anti-inflammatory activity, it might be envisage as an alternate source of staple crop and natural soruce of antioxidant and anti-inflammatory agent.ThesisItem Open Access Biochemical analysis of Buckwheat (Fagopyrum esculentum Moench) genotypes collected from various geographical regions of Uttarakhand(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-07) Rana, Amit Singh; Verma, K.C.Many herbs in the Himalayan region offer nutrition in high value and traditional ways to overcome several diseases. Various food crop offer nutrition of quality and are in use for thousands of years. However, there are still many crops that remain neglected due to the monopoly of some popular crops. Therefore, a search for a crop that is nutritionally high and rich in natural compounds with antioxidant activity has intensified in recent years. In the present study, six genotypes of buckwheat were assessed for their nutritional value, qualitative analysis of seed extract and antioxidant potential. The proximate composition showed variation in value for biochemicals like protein (13.83%), fat (3.31%), starch (68.54%), resistant starch (37.18%) and tannin (2.08%). Bageshwar genotype showed best results for proximate composition. The extraction yield (%) was maximum in Bageshwar genotype in methanol (7.85%) and hexane (3.01%). All the tested phytochemicals were present in all genotypes, in both methanol and hexane extracts. The seed extract analysis by GC-MS reveled active components that show all sort of pharmacological activity from antioxidants to anti-cancerous. The values for antioxidants in genotypes was maximum in Bageshwar genotype (53.71%)in methanol and Rudraprayag genotype (28.47% ) in hexane in terms of DPPH, ferrous chelating activity was highest in Bageshwar genotypes (48.84 %) in methanol and Rudraprayag genotpye (10.24%) in hexane and total antioxidant activity was found to be maximum in Bageshwar genotypes (42.82 mMAA/mg and 08.98 mMAA/mg) for both methanol and hexane extracts, respectively. The total phenol content (mg GAE/g) was maximum in BAG genotype (14.41 and 2.36) in methanol and hexane, respectively while the total flavonoid content (μg CE/g) was maximum in BAG genotype (581.47 and 38.34) in methanol and hexane respectively. Among the solvents, methanol showed comparatively higher antioxidant activity than hexane. Positive and significant correlation in terms of DPPH, FCA, TAA and TPC was obtained in all genotypes. Thus, buckwheat with higher nutritional value, antioxidant potential and a number of other health benefits is comparable to and even sometimes better than other popular crops can be a potent natural alternate staple crop and provide benefits beyond nutrition because of the presence of its phyto-active constituents. It can be exploited in nutraceutical and pharmaceutical industries.ThesisItem Open Access Phytochemical analysis and evaluation of biological activities of Acmella uliginosa (Sw.) Cass. growing in Kumaun region of Uttarakhand(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-07) Gairola, Kanchan; Dubey, S.K.The present work focused on the phytochemical analysis and evaluation of biological activities of Acmella uliginosa (Sw.) Cass. commonly called “Akarkara” growing in Kumaun region of Uttarakhand. The plant Acmella uliginosa, generally growing as weed, was collected from terai region of Uttarakhand and subsequently subjected to hydrodistillation for essential oil. Hexane and methanol extract were prepared with the help of soxhlet. The essential oil and plant extract was chemically analysed by GC/MS. Major compound in essential oil, hexane and methanol was Limonene (12.23%), 2, 4-heptadienal (7.99%) and trans, trans-9,12-octadecadienoic acid propyl ester (16.96%), respectively. In-vitro antioxidant activity of essential oil, hexane and methanol extract were investigated by DPPH free radical scavenging activity, reducing power activity and metal chelating activity of Fe2+. Methanol extract showed maximum free radical scavenging activity with IC50 value 153.82±1.69 μg/ml against DPPH free radicals with respect to ascorbic acid (51.56±0.44 μg/ml) which was used as standard. Essential oil showed better metal chelating activity of Fe2+ and reducing power activity than methanolic and hexane plant extracts having IC50 value of 14.853±0.106 μl and RP50 value of 14.011 ± 0.0446μl. The essential oil showed maximum anti-inflammatory activity with IB50 value of 5.6290±0.0311μl whereas Diclofenac sodium which was used as standard had IB50 value of 23.693± 0.306 μg/ml. Total phenolic content, Total flavanoid content (TFC), Ortho-dihydric phenol content (ODP) and Total antioxidant activity (TAA) of methanol and hexane extract were also evaluated. TPC, TFC, ODP and TAA content was higher in methanol extract having 122.23± 0.22, 35.01± 0.29, 8±0.86 and 111±0.12mg/ml, respectively. The correlation of TPC of plant extract with DPPH radical scavenging activity was -0.99959, with metal chelating activity was 0.999307 and with reducing power was -0.90292. Significant antibacterial and antifungal activity were not observed in methanol and hexane extract. The essential oil showed exceedingly good antiinflammatory as well as antioxidant activities and plant extracts exhibited moderate activity as compared to standard marketed drugs thus, they might be envisaged as an alternative natural source of antioxidant and anti-inflammatory agents.ThesisItem Open Access Studies on pyrolysis products from bio-residues for antimicrobial activity, resin and grease preparation(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-07) Tarannum Jahan; Verma, A.K.Pyrolysis products (bio-oil and the aqueous phase of bio-oil) of pine needles, wheat straw, rice husk and sugarcane leaves were used for antimicrobial activity against plant pathogens (Xanthomonas axonopodia, Xanthomonas chrysanthemum, Erwinia tracheiphila, Ralastonia solanacearum and Pseudomonas fluorescens). Biooils and the aqueous phase of bio-oils showed better antibacterial activity at 1000μg/ml. The pine needles bio-oil (14.00±1mm) and the aqueous phase of bio-oils (10.00±1mm) were showed the highest zone of inhibition among all the bio-oils. Hence, pine needle bio-oil and its aqueous phase could be used as antibacterial agent. The pine needles bio-oil have approximately 50% phenols which can be used for partial replacement of the phenol to synthesize the bio-oil-phenol formaldehyde resin. The shear strength analysis of resin showed that 10% and 20% BPF resin have better bonding strength. The thermogravimetric analysis (TGA) and derivative thermogravimetry (DTG) was showed that the pure PF resin has maximum thermal decomposition as compared to 10% and 20% BPF resins. Scanning electron microscopy (SEM) analysis of resin showed that pure PF resin and 10% BPF resin have almost similar surface structures. Hence, pine needle bio-oil could be used for phenol-formaldehyde resin. The pine needles bio-oils also used for grease preparation. Gear-oil (mineral-oil) was replaced by bio-oil as base oil and animal fat was used as a thickener. The TGA and DTG curves of grease samples were showed that grease sample A (Na-based with mineral oil) having the highest thermal decomposition as compared to sample B (Na-base with bio-oil) and sample C (Li-based with bio-oil). The better dropping point (192ºC) and oxidation stability (136min) were found higher in grease sample C compare to A and B. Cone penetration and copper corrosion values were the same for all the samples showing its liquid consistency and metal protecting nature. The highest value of cone penetration due to the high moisture content in all samples. In grease samples B and C, iron (Fe), Copper (Cu) and Lead (Pd) were found in higher concentration. The additives Ca, Mg, Zn and P were found in very low amount in the grease samples. The ferrographical analysis was found very few wears and contaminations in grease samples (A, B and C), indicating a better behaviour of grease.ThesisItem Open Access Comparative phytochemical profiling of Asparagus racemosus (Safed shatavar) and Asparagus racemosus (Pili shatavar) and assessment for its bioactivity and mineral composition(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2019-07) Leema; Punetha, HimanshuThe aim of present study was to compare phytochemical composition and evaluation of biological activities of pili shatavar and safed shatavar. The extraction was done using methanol as solvent. The extracts was concentrated using distillation. A. racemosus is a biochemical factory of different class of metabolites including sterols, saponins, terpenoids, alkaloids, tannins, glycosides, carbohydrates, flavonoids, coumarine and amino acid. The extracts obtained from soxhlet extraction analysed by combination of GC and GC-MS. The percentage of major compounds present in the root extract of pili shatavar were 1,3:4,6-dimethylene-d-glycero-d-mannoheptitol (22.76), 3-deoxy-d-mannoic lactone (19.43), 5-(hydroxymethyl)-2-furaldehyde (18.91), 2,2-dideutero-trans-1,3-dihydroxy-cyclopentane (12.54), methyl sorbate (7.19), dimethyl fumarate (3.37), oleic Acid (3.24), diisopropyl 2-oxomalonate (2.75) dimethyl maleate (2.21). Whereas the major components present in the root of safed shatavar were 5-(hydroxymethyl)-2- furaldehyde(24.41), curdlan (20.71), pentanoic-3,3-D2 acid (10.09), cyclopentane, 1-acetyl-1,2- epoxy(7.45), oleic acid (4.54), 2-butanone, 4-hydroxy-3-methyl (4.39), pyranone (3.56), tetradecanoic acid (2.54), diethyl methoxymalonate (2.34).The minerals were analysed by AAS. Ca was observed to be highest in pili shatavar 5.276 μg/g. The Mn content was maximum in safed shatavar (0.934μg/g). The iron content observed to be 1.134μg/g in pili satavar and1.172μg/g in safed shatavar. The Zn content was maximum in pili shatavar (0.364μg/g). Cu content was trace amount in pili and safed shatavar. The TPC was maximum in pili shatavar (69±0.012mg/gm) as compared to safed shatavar (36.5±0.33 mg/g). The TFC was maximum in pili shatavar (15.32±0.29 mg/gm) and minimum in safed shatavar (10.09±0.12mg/g). Root extract from pili shatavar showed highest total antioxidant activity (IC50=189.77±4.36) as compared to safed shatavar (IC50=254.57±4.25). The in-vitro antioxidant activity was performed in terms of reducing power, DPPH radical scavenging and metal chelating effect. Pili shatavar methanolic extract (RP50=79.06±1.69) observed to have highest reducing power as compared to safed shatavar (RP50=177.12±4.47). The extract obtained from the pili shatavar displayed good radical scavenging potential with the IC50 value of 536.66±0.92 as compared with the standard antioxidant. Pili shatavar methanolic extract exhibit highest metal chelating effect (IC50=271.21±0.20) in comparison to safed shatavar (IC50=599.78±10.10). However root extract of safed shatavar (IC50=199.19±0.38) observed to have a strong in-vitro anti-inflammatory effect relative to the pili shatavar. The methanolic extracts of pili and safed shatavar have been examined for in-vitro antibacterial activity against two bacterial strains and antifungal activity against three fungal species. The health promoting phytochemical of Asparagus racemosus and their antioxidative potential could be utilized in pharmaceutical sector and food industries.