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Chaudhary Charan Singh Haryana Agricultural University, Hisar
Chaudhary Charan Singh Haryana Agricultural University popularly known as HAU, is one of Asia's biggest agricultural universities, located at Hisar in the Indian state of Haryana. It is named after India's seventh Prime Minister, Chaudhary Charan Singh. It is a leader in agricultural research in India and contributed significantly to Green Revolution and White Revolution in India in the 1960s and 70s. It has a very large campus and has several research centres throughout the state. It won the Indian Council of Agricultural Research's Award for the Best Institute in 1997. HAU was initially a campus of Punjab Agricultural University, Ludhiana. After the formation of Haryana in 1966, it became an autonomous institution on February 2, 1970 through a Presidential Ordinance, later ratified as Haryana and Punjab Agricultural Universities Act, 1970, passed by the Lok Sabha on March 29, 1970. A. L. Fletcher, the first Vice-Chancellor of the university, was instrumental in its initial growth.
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ThesisItem Open Access Studies on biochemical basis for cotton leaf curl disease tolerance in Gossypium hirsutum L.(CCSHAU, 2018) Taranjeet Kaur; Mandhania, ShiwaniViruses 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.ThesisItem Open Access Extraction and evaluation of antioxidant capacity of total phenolics from lychee (Litchi chinensis L.) seeds(CCSHAU, 2018) Nagesh C.R.; Ajay PalLychee 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.ThesisItem Open 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.ThesisItem Open Access Morpho-physiological and biochemical responses of cotton (Gossypium hirsutum L.) genotypes upon sucking insect-pests infestation(CCSHAU, 2017) Vikram Singh; Mandhania, ShiwaniThe 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.ThesisItem Open Access Biochemical and morpho-physiological changes in Indian mustard [Brassica juncea (L.) Czern & Coss.] under terminal heat stress(CCSHAU, 2017) Narender Mohan; Nisha KumariThe 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.ThesisItem Open Access Salt stress induced biochemical changes for delayed leaf senescence in wheat (Triticum aestivum L. em Thell.)(CCSHAU, 2017) Goyat, Ankita; Madan, ShashiThe present investigation was conducted to study the effect of salt stress on physiological, biochemical, yield and quality parameters in two wheat varieties i.e saltsensitive (HD 3086) and salt-tolerant (KRL 210). Salt stress was imposed by irrigating the pots with artificial saline water of 8 and 12 dSm-1. The leaf samples were analysed for various physiological and biochemical parameters at 0, 10 and 20 days after anthesis. The relative water content, chl a and chl ratio gradually decreased with increasing days after anthesis whereas, more decrease in osmotic potential was at 10 days after anthesis in both cultivars and the magnitude of reduction was more in salt-sensitive cultivar (HD 3086) with increase in salt concentration. Chlorophyll stability index was low in HD 3086. The production of malondialdehyde content (MDA), which is an indicator of cell-membrane damage also increased in both cultivars with higher level of increment in the salt-sensitive cultivars than salt-tolerant cultivar. Salt stress resulted in higher increase in the activities of APX and POX in KRL 210 at 10 days after anthesis. Contrarily, ascorbic acid and carotenoids declined in leaves of both cultivars but more in HD 3086 cultivar upon exposure to salt-stress and days after anthesis. Adverse effect of salt stress on plants was associated with accumulation of less K+ and more Na+ in their flag leaf. This resulted in lower K+/Na+ ratio in both the varieties of wheat. Ratio K+/Na+was high in KRL 210. Salinity caused a decrease of hectolitre weight and an increase of protein, gluten and sedimentation value. Salinization led to reduction in test weight, grain number per spike, grain weight per spike, grain yield per plant. Salt susceptibility index (SSI) was less in KRL210.ThesisItem Open Access Optimization of extraction conditions and estimation of antioxidant capacity of phenolics from Jamun (Syzygium cumini L.) seeds(CCSHAU, 2017) Yadav, Neha; Ajay PalDuring present investigation, optimum conditions to extract total phenolic compounds (TPC) from jamun (Syzygium cumini L.) seeds were investigated using one-factor-at-a-time approach followed by response surface methodology (RSM). The central composite rotatable design (CCRD) was used to investigate the effects of three independent variables namely volume of solvent (25-35 ml/g), extraction time (90-150 min), and agitation rate (150-210 rpm) on the response. Regression analysis showed that ~92% of the variation could be explained by the model. The optimized conditions for extraction of TPC were 25 ml/g (50% acetone), 150 min (extraction time) and 193 rpm (agitation rate). The experimental values were in accordance with the predicted values, indicating the suitability of model and success of RSM in optimizing the extraction conditions. Jamun seed extract (JSE) was assayed for its antioxidant activities using an array of tests including free radical scavenging activity, metal chelation activity, hydroxyl radical scavenging activity, hydrogen peroxide scavenging activity, total antioxidant activity (TAA), total reducing power (TRP), nitric oxide scavenging activity (NOSA), lipid peroxidation inhibition activity (LPIA) and protein oxidation inhibition activity. Prokaryotic DNA damage protective efficacy was also observed using pBR322 plasmid. Results of various experiments showed substantial antioxidant activity of JSE and advocate that jamun seed is a good candidate for exploitation as a source of natural antioxidants to counter autoxidation-induced pathologies or diseases.The results of CCRD, used to study the linear, quadratic and interactive effects of temperature, pH and incubation time on the stability of TPC, reveal the existence of significant interaction between temperature and incubation time. A regression equation was successfully developed to deduce the % residual TPC under any conditions of parameters within the experimental domain. The findings of the present study will be of immense help while using JSE as a natural antioxidant in different food systems.ThesisItem Open Access Protective role of spermine on salinity induced oxidative stress in rice (Oryza sativa L.) seedlings(CCSHAU, 2010) Snehvart; Malhotra, SarlaThe effect of salt stress and spermine was studied on oxidative stress and antioxidative system in rice seedlings. The seeds of salt-tolerant (CSR-10) and salt-sensitive (IR-28) cultivars of rice were germinated on Whatman filter paper No. 1 for 4 days and then the seedlings were transferred to hydroponic culture system. The three week old rice seedlings were subjected to 50 and 100 mM salt stress for 10 days. The protective effect of spermine was studied by its exogenous application at 0.1 and 1 mM concentration before salt stress or along with salt stress or after salt stress. The roots and leaves of control, salt-stressed and spermine treated stress plants were analysed for lipid peroxidation (MDA, O2.-, H2O2 content and LOX activity), activities of antioxidative enzymes (CAT, SOD, GR, POX and APX) and antioxidative metabolites (ascorbate and glutathione). Imposition of stress resulted in increase in H2O2, O2.- and MDA content and LOX activity, however, the increase was more pronounced in IR-28. The activities of antioxidative enzymes, in general, decreased in both the tissues of IR-28 but increased in those of CSR-10 upon exposure to salinity. Exogenous application of spermine could partially alleviate the deleterious effect of salinity by lowering H2O2, O2.-, MDA content and LOX activity both in leaves and roots of salt stressed plants of both cultivars. The level of ascorbic acid and reduced glutathione content increased by spermine application to stressed plants of both cultivars. All antioxidative enzymes exhibited an increase in activities in both cultivars after spermine application but the higher concentration (1 mM) had no effect on GR and SOD activity in leaves of CSR-10. Thus, PAs are able to influence oxidative stress intensity by moderating the activities of ROS scavenging enzymes.ThesisItem Open Access Partial purification and characterization of peroxidase from pearl millet [Pennisetum glaucum (L.) R. Br.](CCSHAU, 2010) Goyal, Preeti; Chugh, L. K.This is the first report describing purification and characterization of peroxidase from pearl millet grains. Peroxidase from pearl millet hybrid HHB 94 was purified using ammonium sulphate fractionation, gel filtration chromatography and ion exchange chromatography using Sephadex G - 100 and DEAE cellulose respectively, to near homogeneity. The protocol yielded 41 % of peroxidase with 33.2 fold purification. The purified enzyme preparation has molecular weight of 31 kDa as determined by gel filtration through Sephadex G – 100 and 30.4 kDa by SDS – PAGE suggesting that the enzyme is a monomer. The enzyme exhibited maximum activity at pH 5.6 and 30 °C. It was most stable in the pH range of 6.0 – 6.8 and at temperature below 35 °C. Peroxidase showed Km value of 0.10 mM and 11 mM for o – dianisidine and H2O2, respectively. Activity of peroxidase was highly inhibited by DTT, β – mercaptoethanol , hydrazine and moderately inhibited by sodium azide, sodium borohydride, oxalic acid, EDTA and DTNB. The enzyme activity was enhanced by Ca2+ and Fe3+ but moderately inhibited by Mn2+ , Na+ and K+. Electrophoretic studies of pearl millet grain peroxidase revealed three isozymes in HHB 94 and ICMA 94222 x 78/711. Banding pattern was similar in both but banding intensity represented that ICMA 94222 x 78/711 had lesser peroxidase activity compared to HHB 94.