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ThesisItem Embargo Evaluation of Momordica spp. for bioactive components and antidiabetic potential(Punjab Agricultural University, 2022) Singla, Diksha; Sangha, Manjeet KaurThe present study evaluated antidiabetic potential of bioactive components from immature fruits (60 genotypes) of Momordica spp. in streptozotocin-induced diabetic Wistar rats. The fruit and its parts; epicarp, mesocarp, endocarp, and seeds were evaluated for biochemicals. PAUBG-407 possessed higher phenolics, fiber components, carotenoids, saponins, macro and microminerals; PAUBG-195 (alkaloids); PAUBG-130 (ascorbic acid and tocopherols); PAUBG-218 (proteins) and Punjab-15 (most bioactives). Whole fruit was rich in many bioactives. Charantin, soluble dietary fiber and momordicin-II were purified from whole fruits of PAUBG-407 while vicine and momordicin-I were purified from whole fruit of PAUBG- 195 and PAUBG-89, respectively and characterized on NMR and FT-IR. Their antidiabetic potential was evaluated in 7-8 week old rats, orally administered bioactives @100mg/kg b.w, daily for 4 weeks. Nine groups were made; normal (), untreated diabetic (II), diabetic + (metformin (I), charantin (IV), SDF (V), vicine (VI), momordicin-I (VII), momordicin-II (V), and diabetic+ all bioactives mixed in equal ratio (IX)). Their effect was assessed weekly and at 4 weeks (some parameters). Fasting and random blood glucose; kidney and Iiver CAT, SOD, GPx, GST, GSH. and TBARS: KFT (urea, creatinine, uric acid, Na', K and Ca); LFT (bilirubin. direct and indirect bilirubin, SGOT, SGPT, ALP, total protein, albumin and globulin): plasma insulin, Hb and HbAl c; serum lipid profile (TC, TG, VLDL, DL and LDL), hepatic enzymes- hexokinase, glucose-6-phosphatase and fructose-6- phosphatase; histological aberrations observed in liver, kidney, pancreas and brain improved SIgnificantly (p<0.0001) in diabetic rats treated with all bioactives but highly with momordicin-II and bjoactive mixture. PAUBG-407 may be used in improvement programmes generating antidiabetic karela. Future plans may envisage in testing antidiabetic potential of f targeted bioactives or use of targeted advanced line in diabetic human subjects.ThesisItem Restricted Malt quality and thermostability studies in mutants, hulled, hull-less and wild genotypes of barley (Hordeum vulgare L.)(Punjab Agricultural University, 2023) Heena Rani; Bhardwaj, Rachana DDespite various attempts to develop malt hydrolytic enzymes, the catalytic activity and thermostability of these enzymes are still far from sufficiency for their use in the malting process. The present study explored diverse barley germplasm to identify the genotypes with high activity and thermostability of enzymes alongwith the proteins responsible for high malt yield. Based on enzymatic assays, we selected BL1714T, DWRB137T and BL2105T with highly activity and thermostability of α and β amylases and limit dextrinase, which are key diastatic power enzymes. The barley proteome of these genotypes was analyzed before and after kilning using label-free quantitative global proteomic approach against the control genotypes; BL1718Ctrl, DWRUB64Ctrl and BL2083Ctrl, respectively. Among various isoforms of hydrolytic enzymes Bamy 1 and Amy1_2 were more abundant in thermostable genotypes indicating their role in controlling the activity of these enzymes at high temperatures. Gene ontology enrichment analysis indicated disulfide reductase, exopeptidase, serine-type peptidase (serpin), cell wall hydrolase and oxidoreductase activities as most common enriched terms in thermostable genotypes. These proteins work to release the bound form of hydrolytic enzymes, resulting in their increased overall activity. These proteins could be employed as biochemical indicators for identifying barley lines with high activity and thermostability. Using GC-MS/MS, we identified maltose, glucose, talose, sucrose, fructose, turanose and other minor sugars in green and kilned malt. Talose and turanose were identified among major sugars for the first time during malting. Further, malt quality analysis suggested that βglucanase and malt β-glucan can be used as additional traits to produce malt with superior quality.ThesisItem Embargo Defense response and source-sink relationship for imazethapyr tolerance in lentil (Lens culinaris L.).(Punjab Agricultural University, Ludhiana, 2023) Shivani; Grewal, Satvir KaurPlant growth and yield reduction in lentil crop due to weed infestation is due to poor weed-crop competition. Imazethapyr (IM), controls wide spectrum of weeds as a selective herbicide that targets acetolactate synthase (ALS) which catalyzes the first reaction in biosynthesis of branched chain amino acids (BCAAs), required for plant growth and development. The present investigation was conducted to understand the effect of IM treatment and weed interference on defence mechanism, source-sink relationship and ALS activity along with morpho-physiological traits in IM-tolerant (LL1397 and LL1612) and susceptible (FLIP2004-7L and PL07) lentil genotypes. Lentil genotypes were grown under control (weed free), weedy check (weeds were growing with crop) and sprayed with imazethapyr at 50 days after sowing (DAS). Lower accumulation of methylglyoxal (MG) is due to higher activities of glyoxalase I, II & III enzymes, methylglyoxal reductase (MGR) and lactate dehydrogenase (LDH) in LL1397 and LL1612 after IM treatment as compared to FLIP2004-7L and PL07 along with higher glutathione (GSH) content due to increased γ-Glutamylcysteine synthetase (γ-GCS) help in their recovery post herbicide treatment. Imazethapyr treatment decreased chlorophyll and carotenoid content, plant height and number of pods under IM treatment in FLIP2004-7L and PL07, which resulted in decreased seed yield. Higher activities of ALS, threonine deaminase (TD) with or without any effector molecules and leucine dehydrogenase (LeuDH) was observed in LL1397 and LL1612 for maintaining BCAA pools as compared to FLIP2004-7L and PL07. Expression analysis of ALS gene depicted that 4 days after spray is a critical stage to show its effectiveness at which LL1397 and LL1612 showed higher expression while in PL07 and FLIP2004-7L, expression was highly reduced. Lower H2O2, MDA and DHA content, higher activities of antioxidant and proline metabolizing enzyme activities along with higher ascorbate and proline contents in tolerant (LL1397 and LL1612) might be protecting from herbicide induced oxidative damage thereby improving crop yield. Significantly higher activities of carbon metabolism enzymes in leaves, podwalls and seeds at different developmental stages in tolerant genotypes under herbicide stress as compared to susceptible lentil genotypes might be responsible for maintaining source-sink relationship required for proper growth, grain filling and seed setting. Similarly, enzymes of nitrogen metabolism also showed higher activities in all tissues of tolerant genotypes required for proper nitrogen uptake and better assimilation of inorganic nitrogen into organic nitrogen. Tolerant genotypes had higher total phenolic acid content after imazethapyr treatment as compared to susceptible genotypes with higher content of catechin hydrate, p-coumaric acid, caffeic acid, vanillic acid and rutin that might be contributing towards their better tolerance ability. Soluble protein content was decreased in all tissues of susceptible genotypes as IM treatment affected the protein turnover by accumulating free amino acids. This comprehensive knowledge of mechanisms underlying these metabolic changes after herbicide treatment might provide an efficient way of crop improvement by developing IM-tolerant genotypes.ThesisItem Open Access Biochemical studies on methionine accumulation in maize grains(Punjab Agricultural University, Ludhiana, 2022) Veena Devi; Chaudhary, Dharam PaulPresent study was designed to understand the accumulation pattern of essential amino acids (methionine, lysine and tryptophan), methionine synthesis associated key enzymes activities and responsible genes expression in maize grains. For this purpose diverse panel of 300 inbred lines, procured from ICAR-IIMR and CIMMYT were screened for methionine content and a set of 22 experimental lines (12 high methionine and 10 low methionine) was selected for further study. Field parameters viz., Plant height, cob height, stem and root logging, no. of cobs, no. of rows/cob, no. of seeds/row, germination rate, anthesis and silking intervals, 100 kernel weight, specific gravity, moisture content were recorded. None of the field parameter showed any positive significant correlation with methionine content. The lines E7-24-3, E711-1, E19-14-1, E4-5-2, E2-52-1, E2-7 and E19-22-2 showed better field performance whereas the lines 174705 (3.293%) and 194010 (2.275%) showed highest methionine content thus can be used as acceptor and donors in breeding high methionine maize, respectively. Kernel colour showed correlation with methionine content. Lysine and tryptophan showed declining trend during kernel development and their accumulation was higher in low methionine lines. Aspartate kinase showed similar activity in both high and low methionine lines whereas cystathionine-gamma-synthase does not showed any fixed trend. Serine acetyltransferase enzyme activity was observed to be higher in high methionine than low methionine lines. Higher activity in high methionione lines indicated higher rate of cysteine synthesis, and its flow towards the synthesis of methionine. Its activity showed significant correlation (r = 0.87) with methionine content at 45th DAP. During kernel development zein (prolamin and prolamin-like) accumulated at higher rate in comparison to non-zeins (albumin, globulin and glutelins) in total protein. Methionine showed significant positive correlation with prolamins (r = 0.65) and proteins (r = 0.47). In SDS-PAGE 10-kDa δ-zein was more prominent in high methionine lines, whereas other zeins did not showed any change in high and low methionine lines. Lastly, the mRNA was quantified to study the expression profile of genes responsible for methionine accumulation. It was observed that 10-kDa δ-zein showed enhanced expression in high methionine lines, so its expression affects the accumulation of methionine in developing maize kernel.ThesisItem Open Access Role of γ-amino butyric acid against stripe rust in wheat (Triticum aestivum L.)(Punjab Agricultural University, Ludhiana, 2021) Parmdeep Singh; Zhawar, Vikramjit KaurPBW 725 was found the most resistant cultivar against stripe rust in a screen of seven wheat cultivars under field conditions. GABA applications improved the performance of all seven cultivars against stripe rust. PBW 725 (resistant) and HD 2967 (susceptible) were compared for antioxidant, phenolic, GABA, and carbohydrate metabolisms at 1, 2, and 4 d of GABA, Puccinia striiformis. sp. tritici (Pst), GABA+Pst given to 30-d old seedlings growing in the greenhouse. Fold increase of SOD, APX, GR, GPX, and GST was higher at 1 d in PBW 725 while at 4 d in HD 2967. H2O2 increased at 1 d but decreased at later stages in HD 2967, while PBW 725 maintained H2O2 level during the stress period. PBW725 sustained CAT, DHAR, MDHAR, soluble-POX/PPO, lignin, phenolics, and flavonoids, while HD 2967 became deficient at the late stage Pst. An early increase of lignin was seen in leaves of PBW 725 only. Glucanase and chitinase increased by a significant amount at the late stage of Pst in HD 2967 only. Compared to PBW 725, starch decreased earlier and glucose utilized by more amount in leaves while starch hydrolyzed poorly, and fructose, sucrose, and proline became deficient in roots in HD 2967 under Pst. Compared to PBW 725, glutamate decarboxylase, diamine oxidase, and GABA increased poorly in HD 2967 under Pst. Proteases increased only in HD 2967 at earlier stages but decreased by 4 d of Pst. Protein levels became low by 4 d of Pst in HD 2967 only. Toxicity as hydroxyl, malondialdehyde and protein carbonyls increased only in HD 2967 and raised more in roots than leaves under Pst. GABA improved all studied metabolisms in both cultivars under non-stressed as well as stressed conditions. Toxicity decreased, and defense increased on the application of GABA in HD 2967 under stripe rust. Results concluded that stress response over time is related to resistance against stripe rust. Antioxidant sustainability during the stress period, sugar to starch interconversions, and GABA-shunt activity may be the components of resistance. GABA applications can improve stripe rust resistance in wheat.ThesisItem Open Access Characterization of wheat lines with rye translocation for biochemical and quality parameters(Punjab Agricultural University, Ludhiana, 2021) Ramandeep Kaur; Grewal, Satvir KaurThe replacement of the short arm of wheat chromosome 1B with short arm of rye 1R chromosome (1BL/1RS translocation) has been widely used around the world to enhance wheat yield potential, resistance to rust and mildew and adaptation. However, the translocation is combined with inherent quality problems associated with reduced dough strength and dough stickiness (`sticky dough syndrome') mainly because of two loci on 1RS, presence of Sec-1 on proximal end encoding ω-and γsecalins and absence of Glu-B3 linked Gli-B1 on distal end. The study aimed at using two NILs, one carrying Glu-B3-/Sec-1 and the other carrying Glu-B3+/Sec-1+ from 1BS, generated in background of Pavon (Pavon44:38 and Pavon40:9) for transferring these two loci in yellow rust resistant version of two elite wheat varieties PBW550 + Yr5 (BWL3558) and DBW17 + Yr5 (BWL3279) carrying 1BL/RS translocation (Glu-B3-/Sec-1+) through maker assisted backcross breeding (MABB) for addressing the quality concern of 1BL/1RS translocation lines and their characterization on the basis of biochemical and quality parameters associated with bread baking characteristics. BC1F5:6 advanced backcross introgression lines (ABILs) (Glu-B3-/Sec-1- and Glu-B3+/Sec+1) were generated from four different crosses (Pavon44:38xBWL3558, Pavon44:38xBWL3279, Pavon40:9xBWL3558 and Pavon40: 9xBWL3279) through foreground and background selection. The absence of Secalin protein and presence of GluB3/GliB1 proteins of the selected 126 ABILs were also confirmed through SDS-PAGE. The evaluation of these 126 selected ABILs for yield related traits, biochemical as well as quality characteristics was done for two years. In yield trial, 21 ABILs with Sec-1 and 9 ABILs with Glu-B3 gene in the background of PBW550/DBW17 whose performance was better to recurrent parent were selected. Significant GxE effect was observed across the years. On the other end, from these 126 ABILs the well performed ABILs for biochemical and quality characteristics were selected for product making along with the donor and recurrent parent as checks. Effect of environment was found to be significant on all the biochemical and quality traits of the grains. The ABILs selected for product making were having total phenol content (≤ 2mg/g), total carotenoids content (≥ 3.5), free amino acid content (≥ 1.3), protein content (11-12%), hectolitre weight (72-76kg/hl), grain hardness (10-12kg), phenol reaction score (2.5-3.5), grain appearance score (5-6), sedimentation value (40-50cc), gluten index (90-100),), and gluten content (12-15%). In these selected ABILs there were 9 ABILs with Glu-B3-/Sec-1-1 and 4 ABILs with Glu-B3-/Sec-1- gene configuration on 1RS chromosome. The loaf volume and loaf rise was better of the ABILs in which Sec-1- locus have been transferred which indicated that the presence of Sec-1 is more deleterious than the absence of Glu-B3. The developed two sets of recombinant 1BL/1RS lines (Glu-B3-/Sec-1- and Glu-B3+/Sec+1) can be intercrossed and used for the generation of more improved version of 1BL/1RS cultivars (Glu-B3+/Sec-1-).ThesisItem Restricted Studies on antioxidant and anticancer properties of onion (Allium cepa L.) genotypes(Punjab Agricultural University, Ludhiana, 2020) Inderpal Kaur; Chawla, NeenaThe objectives of the present study were to evaluate the changes in accumulation of antioxidants and the activities of enzymes involved in the synthesis of phenolic compounds in thirty onion genotypes during bulb development stages. The expression of gene (FLS) responsible for the synthesis of flavonol was examined in three selected genotypes, in different plant parts during the bulb development. Study included cytotoxic and apoptotic effects of selected onion extracts on cancer cell lines. Genotypic variation in phenolic content was observed in all onion genotypes during the bulb development. Total phenols, flavonols, odihydroxyphenols, anthocyanin and total antioxidant capacity was observed to be increased from onion bulb initiation stage to bulb maturation stage in most of genotypes, whereas quercetin content was highest reported during bulb developmental stage and then decreased towards maturity. Study showed that enzymes (PAL, FLS and ANS) showed increasing trend from bulb initiation to development stage followed by a decline as the bulb proceeded towards maturation. Correlation and heritability of biochemical parameters were also reported. Three onion genotypes were selected on the basis of phenolic contents and color. Selected genotypes were analysed for the expression study of FLS gene. Study showed maximum relative fold expression of FLS gene in edible portion of plant (leaves and bulb) during bulb development stage. Onion extracts from selected genotypes were prepared in two solvents (ethyl acetate and 80% methanol) to study the potential cytotoxic effects on different cancer cell lines. Combination of ethyl acetate extract of PRO-6 genotype showed significant cytotoxic effects on cancer cell lines.ThesisItem Open Access Manipulation of deamination reactions in the rumen Microorganisms(Department of Biochemistry College of Basic Sciecnes & Humanities, PAU-Ludhiana, 1995) Singh, Kawal Preet; Singh, SudarshanThesisItem Open Access Assembly and secretion of casein micelles in lactating bovines(Department of Biochemistry College of Basic Sciecnes & Humanities, PAU-Ludhiana, 1990) Sodhi, Sandeep; Ahuja, S. P
