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ThesisItem Restricted BIOCHEMICAL BASIS OF HARDSEEDEDNESS IN VIGNA SPECIES(PAU Ludhiana, 2012) Richa Ahuja; Sucheta SharmaThe present investigation was undertaken to study percent hardseededness, seed coat constituents and variation in the biochemical composition of cotyledonary tissue of 10 Vigna species viz., SML832, PAU911, SML668, ML818, MASH218, MASH1008, MASH338, MASH114, RBL1 and RBL6. Seeds from these Vigna species were stored in cloth bags for 180 days at room temperature and the effect of storage period on germinability, vigor index and chemical composition of seeds was studied. The % hardseededness in different Vigna species varied from 14-82% at 24 h and 8-68% after 48 h soaking period. The % hardseededness in different Vigna species decreased with storage of seeds upto 180 days under ambient conditions. The seed coats of hard lines contained higher cellulose and hemicellulose content as compared to those from soft lines. There was non-significant variation in the % neutral detergent fiber, % acid detergent fiber and tannin content in seed coats and total soluble sugars, reducing sugars, phenols and phytate content in the cotyledonary tissue of these Vigna species. However, total lipid content varied significantly. At harvest, the germination % and vigor index values were higher in soft lines as compared to hard lines. The germination % and vigor index values in hard lines increased with the storage period upto 180 days. Lipid peroxidation values increased significantly with storage period in seeds of various Vigna species studied whereas catalase and peroxidase activities decreased. The total lipid content in seeds of all the Vigna species decreased whereas phenolic content increased with the storage period. The results of the present study suggests that although the % hardseededness in different Vigna species decreases with increasing storage period but the seed deterioration occurs with the passage of time.ThesisItem Restricted BIOCHEMICAL CHARACTERIZATION OF WHEAT GENOTYPES WITH RESPECT TO NITROGEN USE EFFICIECNY(PAU Ludhiana, 2012) Gurpreet Kaur; Bavita, AsthirIncreased use of nitrogen (N) fertilizer has raised concerns because N surplus causes environmental contamination and also high cost associated with its production. Thus, improving nitrogen use efficiency (NUE) through identification of efficient genotypes is becoming a necessity. In this two years study (2009-10 and 2010-11), 18 wheat genotypes (PBW 621, PBW 636, PBW 590, DBW 17, HD 2967, PBW 509, BW 9178, BW 9183, BW 8989, BW 9022, PBW 343, PBW 550, GLU 1101, GLU 1356, GLU 2001, GLU 700, PH132-4836, PH132-4840) selected on the basis of their commercial relevance or distinct genetic background were used for studying N metabolism at four N doses including the presently recommended N dose (RDN) (120 Kg N/ha), suboptimal N doses [RDN-50% (60 Kg N/ha) and RDN-25% (90 Kg N/ha)] and supraoptimal N dose [RDN+25% (150 Kg N/ha)]. Enzymes involved in N assimilation [nitrate reductase (NR), nitrite reductase (NIR), glutamine synthetase (GS), glutamate synthase (GOGAT) and glutamate dehydrogenase (GDH)] in relation to deposition of amino acids and proteins were studied at tillering (30 days after sowing; DAS), anthesis (about 90-100 DAS) and post-anthesis (15 days post anthesis) stages. PBW 621 showed higher activities of NR, NIR and GS at RDN-25%. At RDN-50%, PBW 636 and GLU 1356 showed highest GS activity. HD 2967 and PBW 590 revealed maximum activities of these enzymes at RDN and RDN+25%. Widely grown cultivar PBW 343 and advanced breeding lines BW 9178, BW 9183, BW 8989 and BW 9022 showed low efficiency for N assimilating enzymes. GLU 1356 showed high grain protein content indicating higher translocation of assimilates from flag leaf to sink. Sugar and starch content was higher in the PBW 343, BW 9178, BW 8989 and BW 9022 genotypes in which amino acid and protein content was less. PBW 621 showed higher NUE and yield compared to other genotypes. NR and GS enzymes were positively correlated with NUE and yield indicating that these might be the rate limiting steps in N metabolism. Biochemical similarity between PBW 621, PBW 636 and GLU 1356 was authenticated from cluster analysis. Tiller culture technique did not reveal much difference with respect to PBW 621 and PBW 343 in N metabolism. However, hydroponically raised seedlings showed complementary results with field studies in identifying genotypes with maximum NUE. Due to stable performance of PBW 621, PBW 636 and GLU 356 at suboptimal doses over two years, these genotypes hold future potential for developing new cultivars with improved NUE.ThesisItem Restricted Response of sulfur and nitrogen nutrition on storage protein quality in developing seeds of mungbean [Vigna radiata(L.) Wilczek](PAU, 2012) Arvind Kumar; Sharma, SuchetaThe present study was conducted to evaluate the response of sulfur and nitrogen nutrition on storage protein quality in developing seeds of mungbean [Vigna radiata (L.) Wilczek] varieties PAU 911 and ML 818. Crops were sown in the experimental fields of Department of Plant Breeding and Genetics, Punjab Agricultural University, Ludhiana following recommended package of practices. Before sowing, different treatments of sulfur and nitrogen @40kg/ha were given to the fields i.e. T 1 (control), T2(single super phosphate), T3 (urea), T4 (single super phosphate and urea), T5 (gypsum), T6 (ammonium nitrate) and T7 (gypsum and ammonium nitrate). Maximum biomass accumulation was observed at 16 DAF and maximum dry weight at 20 DAF in both the varieties. Crude protein, total soluble protein, sulfur containing amino acids (methionine and cysteine) contents in developing seeds increased maximally with T5 as compared to control and other treatments. The free amino acids increased upto 20 DAF then declined towards maturity. The albumin and globulin content in developing seeds increased with various treatments of nitrogen and sulfur sources. Among the legumins and vicilin, legumin showed better response to sulfur and nitrogen nutrition, which is desirable as legumin are rich in sulfur containing amino acids. Nitrate reductase (NR), glutamate dehydrogenase (GDH), O-Acetyl serine thiol lyase (OSTL) and glutathione reductase (GR) activities were highest at 18 DAF and then declined towards maturity. NR and GDH activities were maximally increased with urea (T 3) whereas gypsum (T5) showed higher increase in OSTL and GR activities as compared to other treatments in both the varieties. Highest incorporation of C 14 leucine in albumin and globulin protein fraction of developing seeds at 16 DAF was due to gypsum treatment after 12 h and 24 h incubation periods in both the varieties. In mature seeds, gypsum (T5) gave maximum response towards protein quality parameters i.e. total sulfur, total soluble protein, sulfur containing amino acids and seed storage protein fractions. Electrophoretic studies revealed more distinct and intense banding pattern of globulin with T5 as compared to control and other treatments.ThesisItem Open Access Biochemical mapping of soybean (Glycine max (L.) Merrill) germplasm for bioactive compounds(PAU, 2012) Goyal, Reeti; Sharma, SuchetaThe present investigation reports the physicochemical properties and biochemical diversity in contents of nutrients, antinutrients and bioactive compounds in 140 soybean genotypes (74 local genotypes, 51 otypes from other parts of India and 15 exotic genotypes). Soybean genotypes exhibited 38.4-46.5% protein, 20.8-23.6% oil, 1.3-13.9% total soluble sugars, 0.24-13.8% sucrose, 2.50-33.5 mg/100g tocopherols, 8.9- 0.5 mg/g tannins, 11-38.3 mg/g phenols, 1.2-28.5 mg/g phytate and 11.3-142.5 mg/g Trypsin Inhibitor ctivity (TIA). The activities of lipoxygenases (LOX) were in the range from 242.5-2510.3 units/g for LOX-I and 05.1-661.75 units/g for LOX-II+III isozymes. Following screening studies on 140 soybean genotypes in 2009, 20 selected genotypes were sown in year 2010 at two different locations ie. Ludhiana and Gurdaspur and two different planting times viz. June and July with the aim to study the environmental effects on levels of nutrients, antinutrients and bioactive compounds significant effects of genotype, planting time, location and their interactions were observed on physicochemical characteristics, nutrients, inutrients and bioactive compounds of soybean genotypes grown in different environments. Gas Liquid hromatographic studies revealed significant variation for individual fatty acids among these genotypes. PLC analysis of different tocopherol isomers revealed highest contents of α, γ, δ and total tocopherols uring une planting both at Ludhiana and Gurdaspur. Although similar banding patterns of different subunits of β-conglycinin (7S) and glycinin (11S) storage proteins among 20 soybean genotypes were observed in electrophoretic studies but these genotypes showed differences in relative proportions of various bunits of 7S (α’,α and β) and 11S (acidic and basic) proteins in ensitometric analysis. Genotypes ‘SL 989’, ‘SL 992’ and ‘PK 1026’ exhibited higher protein, total sugars, sucrose and tocopherol content and omparatively lower contents of various antinutrients as compared to their mean values for 140 genotypes. These genotypes also exhibited low TIA, phytate and saponin content and higher protein, tocopherol content and seed yield across both locations and planting times. Genotypes ‘SL 313’, ‘SL 790’ and ‘YMV-35’ exhibited higher oil, 18:2 and 18:3 and lower 18:1 content as compared to other enotypes. Due to their stable performance across different environments, most of the selected genotypes can be considered important for developing new cultivars with improved nutritional quality traits.ThesisItem Open Access TOXIC EFFECTS ON FISH INHABITING ARSENIC CONTAMINATED FRESH WATERS OF PUNJAB(Punjab Agricultural University, Ludhiana, 2012) Saini, Amandeep Kaur; Hundal, S.S.High concentration of arsenic in groundwater of south-western provinces of Punjab, India has become a major cause of concern. Monitoring arsenic levels and their associated health effects in fish would not only provide an insight into overall ecosystem health but may also act as a sentinel for potential impacts on human health. Freshwater fish was captured from natural sites with known constant elevated levels of arsenic of 12-13 μg/L and brought to laboratory with an objective to analyze fish muscle tissue for arsenic and the chronic effect of elevated levels of arsenic on Hepatosomatic index (HSI), Gonadosomatic index (GSI) and histology of fish. The variation in HSI and GSI values for fish from affected and control sites were statistically insignificant. Mean arsenic concentration of arsenic in the muscle tissue of fish from the area with an elevated level of arsenic was recorded as 172.3±22.4 μg/g as compared to average value of 93.3±13.1 μg/g in control. Histological studies revealed degenerative changes in liver, congestion of blood vessels in kidney along with vacuolar degeneration; besides this, significant difference in the diameter of proximal convoluted tubules and its lumen for kidney tissue was observed. The gonadal changes included the loss in shape of seminiferous tubules in testis and exfoliation of ovarian cells. The histomorphometric observations indicated an insignificant difference in the diameter of the secondary oocyte, while the primary oocyte diameter had significant variation (p>0.0001) from control specimens. These observations are in accordance with earlier studies on the effect of environmental contaminants, but being one of the first reported on fresh water fish exposed in natural waters, it assumes significance. The observations have been related to the physiology as also to the reproductive health of fish and may have a bearing on mammalian health under constant exposure.ThesisItem Open Access Biochemical changes in rice (Oryza sativa L.) genotypes in response to different transplanting time(Punjab Agricultural University, Ludhiana, 2012) Sharma, Kanu PriyaThe present study was conducted to understand the biochemical changes in rice (Oryza sativa L.) genotypes viz., heat tolerant (N22), moderately heat susceptible (IR8) and fine grain rice variety (PR116) in response to different transplanting time. Constitutive level of peroxidase (PO), phenylalanine ammonia lyase (PAL), tyrosine ammonia lyase (TAL), superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) was found to be significantly higher in N22 than in IR8 at very early (D1) and early transplanting (D2) times. Heat stress caused a significant increase in the level of the phenolics constituents and the activities of PO, PAL and TAL in N22. In contrast, the hydrogen peroxide (H2O2) and malondialdehyde (MDA) content was maximum in IR8 and was found to be the highest at D1. Total free amino acid and proline content was significantly higher in N22 than in PR116 and IR8. Total sugars and reducing sugars decreased at D1 and maximum content was found in IR8. The starch content was found to be maximum in N22. Total chlorophyll, chlorophyll a and chlorophyll b content decreased with heat stress and minimum content was present in PR116. Total amino acid content showed an increase with increase in heat stress in IR8 and PR116. No difference was noticeable in the electrophoretic pattern of protein bands in all the three cultivars. The 1000-grain weight of paddy, brown rice and milled rice decreased at D1 and D2, particularly in IR8. Grain dimensions were unaffected by different transplanting dates. A significant decrease was observed in the head rice recovery at D1 and D2 in all the three cultivars with the maximum head rice recovery being obtained at normal transplanting time. Higher expression of PO, PAL, TAL, SOD, GR, CAT, APX, ascorbate content and phenolic constituents in N22 than in the IR8 and PR116 is suggestive of their significant role in development of tolerance exhibited by the cultivar N22 under heat stressed conditions, during D1.ThesisItem Open Access ANALYSIS OF ANTIOXIDANT METABOLISM IN WHEAT SEEDLINGS EXPOSED TO WATER STRESS AND UNDER THE INFLUENCE OF EXOGENOUS ABSCISSIC ACID AND REACTIVE OXYGEN SPECIES(Punjab Agricultural University, Ludhiana, 2012) Manpreet KaurThe present work was aimed to study the involvement of superoxide anions and ABA biosynthesis in germination, growth and antioxidant response under water stress (WS) and under ABA (A) supply in two wheat cultivars C306 (ABA higher sensitive-cum-drought tolerant) and PBW343 (ABA lesser sensitive-cum-drought susceptible) by using tiron (T) as superoxide scavenger and sodium tungstate (ST) as inhibitor of ABA biosynthesis. Biomass and ascorbate were decreased on scavenging superoxide ions under A in both cultivars. Antioxidant enzymes, H2O2 were decreased in PBW343 but increased in C306 on scavenging superoxide ions under A. ABA inhibitions of shoot length, root length and germination were higher in C306 than in PBW343 and these inhibitions were relieved significantly on scavenging superoxide ions in C306 not in PBW343. Fresh biomasses, antioxidant enzymes, ascorbate/dehydroascorbate ratios were decreased and dehydroascorbate and malondialdehyde contents were increased on inhibiting ABA biosynthesis and on scavenging superoxide anions under water stress in both cultivars but such changes were more significant in C306 than in PBW343. Both cultivars were also compared under H2O2 (H) supply for antioxidant response and for the involvement of ABA biosynthesis under H supply. H2O2 induced antioxidant levels were higher in C306 than in PBW343. On inhibiting ABA biosynthesis under H supply, these H2O2-mediated effects were affected but not completely removed in both cultivars.ThesisItem Open Access BIOCHEMICAL AND NUTRITIONAL CHARACTERISTICS OF GREEN AND MATURE SOYBEAN [Glycine max (L.) Merr,] GENOTYPES AT DIFFERENT DEVELOPMENTAL STAGES(Punjab Agricultural University, Ludhiana –, 2012) Kaur, Shaminder; Saxena, A. K.Ten soybean [Glycine max (L.) Merr,] genotypes along with local check (SL-688) were evaluated for physico-chemical, nutritional and processing characteristics at R6 and R8 stages. Percent crude protein, crude fibre, ash content, crude fat and total carbohydrate were observed in the range of 6.13-15.25, 2.24-3.69, 1.10-2.81,7.76-9.82, 14.9-18.03 at R6 and 36.62-43.72, 3.12-5.16, 4.03-4.74, 9.72-17.12, 28.41-35.56 at R8, respectively. SL-744 was found to be most promising w.r.t. methionine and tryptophan content. Considerable amount of antinutrients was reported in soybean which decreases its nutritional quality. Genotypes exhibited the average contents of antinutrients (mg/g) viz. phenols, ortho dihydroxyphenols, tannins, trypsin inhibitor activity, amylase inhibitor activity, saponin and phytate as 11.56, 3.56, 7.26, 21.83, nil, 1.19, 6.10 at R6 and 31.25, 10.96, 18.39, 52.17, 11.99, 2.96, 15.29 at R8 respectively. Genotypes SL-525×AGS328, SL-900 and SL-958 registered lower antinutritional content and hence promising w.r.t. nutritional quality. Soaking, cooking and blanching led to a considerable reduction in phenols, tannins, trypsin inhibitor activity and other water soluble antinutrients. Amongst various methods studied, autoclaving in distilled water was found to be the most effective in reducing levels of antinutrients in soybean genotypes. Protein profile (SDS-PAGE) of soybean genotypes did not register any difference in banding pattern; however variation was observed in the intensity of bandsThesisItem Open Access EVALUATION OF BRASSICA SPP. GENOTYPES FOR MINERALS AND OTHER BIOMOLECULES IN EDIBLE LEAVES(Punjab Agricultural University, Ludhiana, 2012) Khatri, PriyankaBrassicae crops have been related to the reduction of risk of chronic diseases such as cardiovascular diseases and cancer. The beneficial effects of brassicaceae vegetables on health improvement have been attributed to their complex mixture of phytochemicals possessing antioxidant activity. The present investigation was undertaken in different genotypes of B. juncea (4) and B. napus (4) at three different developmental growth stages i.e. (35, 55, 75 days) to evaluate for presence of antioxidants and other nutritionally important biochemical parameters. In B. juncea PBR 91 and RLM 619 genotypes and in B. napus CNH-9-15 and GSL 1 genotypes showed higher levels of antioxidants (tocopherol, Vitamin C and carotenoids). Higher levels of minerals content were found at initial growth stage i.e. 35 days which decreased with growth stage in both B. juncea and B. napus. Maximum iron content was observed in RLC 1 of B. juncea and CNH-9-19 of B. napus whereas zinc levels were maximum in RLM 619 and PBR 91 genotypes of B. juncea and CNH-9-15 and CNH-9-19 of B.napus. Levels of phytic acid were comparatively low in B. napus as compare to B. juncea. Correlation studies indicated a negative correlation between phytic acid and iron and a positive correlation was observed between tocopherol and protein content whereas glucosinolate and phenol content showed a negative correlation at 35 days old leaves in both genotypes of B. juncea and B. napus.
