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2010 - 201934
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Subject: Biochemistry × End date: 2019 × Start date: 2010 ×
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Now showing 1 - 9 of 34
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    ThesisItemOpen Access
    Biochemical studies on chromium toxicity and its amelioration in sorghum (Sorghum bicolor L.)
    (CCSHAU, Hisar, 2019-10-31) Praveen Kumar; Singal, H.R.
    The main aim of the present research work was to determine the effects of Cr (VI) toxicity (2 ppm & 4 ppm) and its amelioration by glycine betaine (GB) and AMF at two growth stages viz. vegetative stage (35 DAS) and grain filling stage (95 DAS) in three varieties of sorghum viz. HJ 541 (single-cut), HJ 513 and SSG 59-3(multi-cut). The effect of exogenous application of GB (spiked in soil & foliar spray) and AMF (10 g) either individually or in combination was investigated for their ameliorative capability. The ameliorative effects were studied in terms of biochemical parameters (viz. structural and non-structural carbohydrates, crude protein, chlorophyll, chromium, hydrocyanic acid, IVDMD, pectin, PME, PG and cellulase), morphophysiological parameters (viz. root length, shoot length, number of leaves/tillers, grain yield, plant height and plant biomass), antioxidative defense system parameters (viz. enzymes – SOD, APX, CAT, GR, POX and metabolites – proline, glutathione, ascorbate, β-carotene) and indices of oxidative stress parameters (viz. PPO, H2O2 and MDA). The results revealed that the biochemical and morphophysiological parameters (except hydrocyanic acid, chromium, non-structural carbohydrates and pectin) decreased with increasing concentration of Cr (VI) in all the varieties at both the growth stages, and maximum decrease was observed in HJ 541 variety at 4 ppm concentration of Cr (VI) as compared to HJ 513 and SSG 59-3 variety at both growth stages. The antioxidative defense system parameters (enzymes & metabolites) and indices of oxidative stress parameters increased with increasing concentration of Cr (VI) at both the growth stages, in all the varieties. Further increase in antioxidative defense system parameters on exogenous GB (spiked in soil & foliar spray) and AMF application, either individually or in combination ameliorated the toxicity and decreased indices of oxidative stress parameters and increased IVDMD, total sugars, crude protein, β-carotene and nutritionally quality of sorghum in all the varieties at both the growth stages. The variety SSG 59-3 had lower hydrocyanic acid, non-structural carbohydrates, chromium, indices of oxidative stress parameters and more IVDMD and nutritional quality parameters as compared to HJ 513 and HJ 541 variety, at both the growth stages. Thus, among the three varieties (viz. HJ 541, HJ 513 and SSG 59-3), SSG 59-3 variety was found more tolerant as compared to HJ 541 and HJ 513 variety.
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    ThesisItemOpen Access
    Effect of phosphorus and potassium application on quality of sorghum (Sorghum bicolor (L.) Moench)
    (CCSHAU, 2019) Seema; Singal, H.R.
    Sorghum (Sorghum bicolor (L.) Moench) locally known as Jawar or Chari is an important kharif season crop which is grown for both fodder and grains. Fertilizers constitute an integral part of improved crop production technology. Application of optimum quantity of phosphorus and potassium is important in improving quality and yield of forage sorghum. The present investigation was carried out to study the effect of phosphorus and potassium application on quality parameters. Sorghum variety HJ 541 was grown in field and different doses of phosphorus (15, 30. 45, 60 and 90 kg P2O5/ha) and potassium (20, 30, 40, 50 and 60 kg K2O/ha) were given. Plant/leaf samples were collected at 30 DAS, 50 DAS and 90 DAS. Biochemical parameters namely HCN, IVDMD, crude protein, nitrogen content, phosphorus content, potassium content, iron content, NDF, ADF, total soluble solids, total soluble sugars, reducing and non- reducing sugars, morpho- physiological parameters namely fresh weight, dry weight, stem girth, internode length and number of leaves and cell wall components namely cellulose, hemicelluloses, lignin and pectin were studied. It was observed that HCN content was less than permissible value (200 μg/g at fresh weight) with the application of phosphorus and potassium. IVDMD, crude protein, nitrogen content, phosphorus content, potassium content, iron content, total soluble solids, total soluble sugars, reducing and non- reducing sugars increased with the application of phosphorus and potassium. Maximum fresh and dry weight, stem girth internode length and number of leaves increased with the application of phosphorus and potassium. NDF, ADF, cellulose, hemicelluloses, lignin and pectin decreased with the application of phosphorus and potassium. The results obtained revealed that the quality of forage sorghum improved with the application of phosphorus and potassium.
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    ThesisItemOpen Access
    Studies on biochemical basis for cotton leaf curl disease tolerance in Gossypium hirsutum L.
    (CCSHAU, 2018) Taranjeet Kaur; Mandhania, Shiwani
    Viruses have profound effect on the growth and productivity of plants. With reference to the cotton crop, CLCuD caused by CLCuBuV has been recognized as a serious threat to cotton production in Indian subcontinent. Information on cotton and CLCuV interaction is still not convincingly explained. In the present study, the constituents like RWC, electrolyte leakage, total chlorophyll content, MDA content, total soluble protein, total sugar content, total phenolics, H2O2, gossypol content and the activities of defense related proteins viz. POX, PPO, CAT and PAL were scrutinized at different stages of infestation in GCH 3, HS 6 genotypes of cotton and in additional genotype (HD 432) to look over the biochemical basis for CLCuD tolerance mechanism. It was found that the maximum RWC, total chlorophyll content and total soluble protein was present in highly tolerant and resistant genotype respectively at 0 DAI. Decline in the amount of these constituents reported in all genotypes but maximum loss took place in HS 6. Least amount of electrolyte leakage, H2O2 and MDA content was present in HD 432 at all stages of infestation. Significant increase in these parameters observed in HS 6 post incursion of virus. Maximum total sugar content was present in HS 6 at 0 DAI, ebb was more in GCH 3 due to infestation. As a consequence of viral infection, the level of total phenolic compounds and gossypol content increased significantly in all genotypes while maximum increase was in GCH 3. With the progression of infestation period significant increase was reported in the activities of PAL, PPO, POX and CAT in all genotypes. Thus, the increase in the level of total phenolic compounds, gossypol with increase in the activities of all studied enzymes can be correlated with the potency of the plant to strive against CLCuD.
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    ThesisItemOpen Access
    Extraction and evaluation of antioxidant capacity of total phenolics from lychee (Litchi chinensis L.) seeds
    (CCSHAU, 2018) Nagesh C.R.; Ajay Pal
    Lychee seed, being a biowaste, is an inexpensive source of a variety of polyphenols. However, extraction yield/efficiency of total phenolics from lychee seed powder (LSP) depends on extraction conditions. Hence, in present study, optimum conditions for extraction of phenolics from LSP were deduced using one-factor-at-a-time approach followed by response surface methodology (RSM). The factorial, axial and center point experiments of central composite rotatable design (CCRD) of RSM were carried out to investigate the linear, quadratic and interactive effects of three independent variables namely volume of extractant (ml/g), agitation rate (rpm) and extraction time (min) on the yield of phenolics. Regression analysis showed that ~92% of the variation could be explained by the model. Software mediated numerical optimization was done with the constraint of minimum volume of extractant and extraction time separately for maximum yield of TPC. The experimental values were in accordance with the predicted values, indicating the adequacy of model and success of RSM in optimizing the extraction conditions. Lychee seed extract (LSE) was analyzed for profiling of phenols and fatty acids using H-UPLC and GC, respectively. Despite reported phenolic compounds, we found new phenolics namely apigenin, catechol, catechin, chlorogenic acid, diadzein, salicyclic acid, trans-ferulic acid, trans-sinapic acid and vanillin in LSE. Among six different fatty acids identified, oleic acid and linoleic acid were abundant. Further, LSE was assayed for its antioxidant activities using an array of tests including free radical scavenging activity, nitric oxide scavenging activity (NOSA), hydroxyl radical scavenging activity, metal chelation activity, total antioxidant activity (TAA), total reducing power (TRP), lipid peroxidation inhibition activity (LPIA) and protein oxidation inhibition activity. Prokaryotic and eukaryotic DNA damage protective efficacy of LSE was also observed using pBR322 plasmid and human lymphocytes, respectively. Results of various experiments showed substantial antioxidant activity of LSE and advocate its exploitation as a source of natural antioxidants to counter ROS-induced pathologies or diseases.
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    ThesisItemOpen Access
    Physico-chemical and kinetic properties of fatty acid esterase from pearl millet [Pennisetum glaucum (L.) R. Br.] hybrid HHB 197 and its parents
    (CCSHAU, 2018) Sheenu; Chugh, L.K.
    Fatty acid esterase (FAE) (E.C. 3.2.2.1) was partially purified from flour of pearl millet hybrid HHB 197 and its parents viz., A-line ICMA 97111 and R-line HBL 11 by ammonium sulphate fractionation (30-60 % saturation) followed by gel filtration through Sephadex G-75. FAEs of A-line ICMA 97111 and R-line HBL 11 eluted as a single peak were designated as FAE I and the enzyme resolved into two distinct peaks from the hybrid HHB 197 were designated as FAE I and FAE II. After gel filtration 15 to 55 % of the enzyme was recovered with specific activity of 7.35 to 12.85 units/mg proteins with fold purification ranging from 2.8 to 5.5. Physico-chemical and kinetic properties of the enzyme of these genotypes were compared. The FAE I of the hybrid HHB 197, A-line ICMA 97111 and R-line HBL 11 had identical molecular weight of about 67.6 kDa. Molecular weight of FAE II of the hybrid was 22.3 kDa. Both FAE I and FAE II of the hybrid showed optimum temperature of 40oC. FAE I of A-line ICMA 97111 and R-line HBL 11 had optimum temperature of 40-45 and 45oC, respectively. FAE I of ICMA 97111 lost maximum activity when incubated for 20 min at 50 OC. At this temperature FAE I of the hybrid was comparatively more stable. Except FAE II of HHB 197, FAE I of all the genotypes showed optimum activity at pH 8. Ascorbic acid, EDTA and SDS were found to be inhibitory for all the FAEs. Fe3+, Cu2+ and Zn2+ showed significant deleterious effect on the activity of FAEs of all the genotypes. Co2+ was stimulatory for FAE II of hybrid HHB 197 and FAE I of R-line HBL 11 and had slight deleterious effect on FAE I of the hybrid and A-line ICMA 97111. All FAEs showed typical hyperbolic response to concentrations of the substrate para-nitrophenylbutyrate (pNPB). Km values were calculated using the software GraphPad Prism ver. 7. Km value of FAE I of R-line HBL 11 (0.540 M p-NPB) was the highest followed by those of FAE I (0.155 M p-NPB) and FAE II (0.100 M p-NPB) of the hybrid and that (0.067 M p-NPB) of A-line ICMA 97111. High Km value of FAE I of the R-line HBL might be partly responsible for slow in situ lipids hydrolysis.
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    ThesisItemOpen Access
    Morpho-physiological and biochemical responses of cotton (Gossypium hirsutum L.) genotypes upon sucking insect-pests infestation
    (CCSHAU, 2017) Vikram Singh; Mandhania, Shiwani
    The four cotton genotypes viz., Bio-100 BG II, GCH 3, KDCHH-9810 BG II and HS 6 were evaluated under field conditions to compare their tolarance levels against the sucking insect-pests. The effect of sucking insect-pests on morpho-physiological biochemical constituents viz., the relative water content, specific leaf weight, total chlorophyll content, electrolyte leakage, leaf area, photosynthetic rate, total phenolic content, total soluble sugar, tannin content, total soluble protein, gossypol content, H2O2 content and activity of enzymes- peroxidase and polyphenol oxidase that could explain some of the varietal tolerance levels were tested. Morpho-physiological and biochemical analysis indicated that the highest relative water content, total chlorophyll content, leaf area and photosynthetic rate was observed in Bio 100 BG-II, highest specific leaf weight was observed in GCH-3 and highest electrolyte leakage was observed in HS-6 genotypes. The lowest relative water content, total chlorophyll content, leaf area and photosynthetic rate was observed in HS-6, lowest electrolyte leakage was observed in Bio 100 BG-II and lowest specific leaf weight was observed in KDCHH 9810 BG-II genotypes. The highest total phenolic content, tannin content and POX activity observed in Bio-100 BG II. Gossypol content, total soluble protein and PPO activity were highest in GCH 3. Total soluble sugar and H2O2 content was highest in KDCHH- 9810 BG II. Total phenolic content, gossypol content and tannin content were lowest in HS 6. Total soluble protein and PPO and POX activity were lowest in KDCHH-9810 BG II. Total soluble sugar was lowest in GCH 3. H2O2 content was lowest in Bio-100 BG II. Sucking pest infestation reduced the total soluble sugar and total soluble protein of all genotypes, while other biochemical constituents increased with sucking insect-pests infestation. These results indicated that Bio-100 BG II is highly tolerant as compare to other genotypes.
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    ThesisItemOpen Access
    Biochemical and morpho-physiological changes in Indian mustard [Brassica juncea (L.) Czern & Coss.] under terminal heat stress
    (CCSHAU, 2017) Narender Mohan; Nisha Kumari
    The present investigations were undertaken to study the changes in oxidative stress, antioxidative enzymes, metabolites and yield & yield attributes in Brassica juncea after subjecting to terminal heat stress at prematurity stage. Two thermo-tolerant genotypes viz. RGN-368 and RH-1566 and two thermo-sensitive genotypes viz. RH-1134 and RH-0749 were sown on two different dates (13th October and 15th November). Hydrogen peroxide (H2O2), malondialdehyde (MDA) and electrolyte leakage which are indicators of cell membrane damage increased under terminal heat stress in all the genotypes but increase was higher in sensitive genotypes. The activities of antioxidative enzymes viz. superoxide dismutase (SOD), peroxidase (POX), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR) increased under terminal heat stress showing that the antioxidant defence mechanism play major role in heat stress tolerance in Brassica juncea. Ascorbic acid, proline and carotenoids increased under heat stress depicting their role as osmoprotectants and free radical scavangers, respectively. Relative water content (RWC), osmotic potential (OP), chlorophyll content and crude oil content was observed and a remarkable decline was reported in these attributes under late sown condition. Yield attributes, such as main shoot length, number of primary and secondary branches, numberof siliqua on main shoot, seeds per siliqua, siliqua length, 1000 seed weight and seed yield per plant also showed a significant reduction under terminal heat stress.
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    ThesisItemOpen Access
    Use of molecular markers for varietal identification and assessing phylogenetic status of Basmati rice in genus Oryza sativa L.
    (CCSHAU, 2011) Jyoti; Jain, Sunita
    Molecular markers provide novel tools for varietal identification, diversity analysis and assessing phylogenetic relationships among various rice groups in genus Oryza. A set of 50 rice genotypes comprising of seven japonica rice varieties, six traditional Basmati, thirteen cross-bred Basmati and 24 indica rice varieties was investigated using 54 markers (28 on chromosome 8 and 26 on rest of the chromosomes); data obtained has been used to assess the phylogenetic status of Basmati rice in genus Oryza sativa L. A total of 217 alleles were detected, with an average of 4.01 alleles per locus. Number of alleles per locus (3.5 alleles) for markers on chromosome 8 was less than the mean value (4.58 alleles) based on 26 SSR markers on rest of eleven chromosomes of rice. Eleven of these alleles were unique, present in only one genotype. Null allele was observed at only one locus (RM408) in Super Basmati. The number and size of alleles and Polymorphism Information Content (PIC) values ranged between 2-7, 83-381 bp and 0.074-0.810, respectively. An average PIC of 0.55 per locus was obtained, which confirms that markers used in this study were highly informative. Genetic relationships generated using the whole genome marker data as well as allelic profile on chromosome 8 and rest of eleven chromosomes separately placed 50 rice genotypes in three distinct clusters. Seventeen of the nineteen Basmati rice varieties (except two cross-bred Basmati rice varieties, Sabarmati and Improved Sabarmati) formed a separate cluster quite distinct from the indica and japonica rice clusters. Interestingly, mean similarity indices obtained from chromosome 8 dataset placed Basmati rice cluster at equal distance from japonica and indica rice types. However, whole genome as well as rest of the genome marker datasets placed Basmati group closer to indica rice varieties than japonica. Chromosome 8 dataset showed a positive correlation (Mantel test, r = 0.631) with the rest of the genome dataset, indicating a higher level of similarity between the two. All the traditional and crossbred Basmati rice varieties and two aromatic japonica varieties shared the same BAD2 alleles at the aroma locus, indicated the distinctness of the aroma locus. While the present study gives support to the hypothesis that japonica genotypes may have contributed towards the evolution of chromosome 8 or a part of chromosome 8 in Basmati rice, it also indicate the active flow of genes from indica rices during the course of parallel evolution in northern Indian sub-continent. The study also demonstrates the distinctness of TB from other rice types (indica and japonica) and also provides a number of novel SSR markers which can be used to differentiate within/among the various rice groups at commercial level. It was observed that the amplified products differing in ≥12bp can be separated on 3% agarose gels for varietal identification in Basmati rice, which is faster, much cheaper and relatively simple method compared to the advanced molecular methods already available for the purpose.
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    ThesisItemOpen Access
    Biochemical changes in tomato (Solanum lycopersicum L.) under the influence of salicylic acid
    (CCSHAU, 2017) Naresh Kumar; Singal, H.R.
    The present investigation was aimed to study the effect of exogenous application of salicylic acid on two tomato varieties Hisar Arun (Short shelf life) and BSS-488 (Long shelf life). Tomato fruits of both varieties were harvested at mature green and turning stage and treated with the salicylic acid at different concentrations (0.5 mM, 0.75 mM, 1.0 mM, 1.25 mM and 1.5 mM). The fruit sample was analysed for various morpho-physiological and biochemical parameters at every 3rd day after salicylic acid treatment. The molecular parameters were analysed only at optimized concentration of salicylic acid (i.e. 0.75 mM) afterwards. The morpho-physiological and biochemical parameters like PLW, TSS, color change, lycopene, β-carotene, PG and cellulase increased progressively while the parameters like cellulose, hemicellulose, lignin and pectin decreased gradually throughout the storage period which leads to the ripening of tomato. The exogenous application of salicylic acid mainly at concentrations 0.75 mM improved the shelf life of tomato by delaying the change in the above mentioned morpho-physiological and biochemical parameters. Moreover, during ripening the parameters such as total sugars, reducing sugars, non-reducing sugars, ascorbic acid, ethylene, PPO and PME activity increased initially and declined thereafter with advanced ripening in control as well as in all treated fruits. However, the treated fruits at SA concentration 0.75 and 1.0 mM expressed the delay in the initial increase in above parameters. Similarly, the expression of ripening related genes (ACS and ACO) of tomatoes of both varieties at both maturation stages increased initially and decreased thereafter which is delayed by the exogenous application of salicylic acid. In addition, the quantitative analysis (via PD Quest) of 2-DE gels of tomato proteins demonstrated the overlapping gel coordinates between control and treated fruits. But the treated fruits expressed the differentially expressed protein spots that showed >1.5 fold expression change as compared to control. So it can be concluded that the treatment at 0.75 mM concentration of salicylic acid was most effective in enhancement of shelf life of tomato fruits.