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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 Study of iron deficiency induced physiological and biochemical responses in wheat seedlings(CCSHAU, Hisar, 2023-01-02) Sindhu, Sudeep; Vinod KumarIron (Fe) is the most critical micronutrient needed for plant growth and development. It is required as a redox active metal that is involved in a variety of physiological and metabolic processes. There is a lack of iron in approximately one third of the world's soil, which hinders the growth and development of plants, which in turn results in lower yields and worse quality nourishment. A lack of iron in plants is difficult to control with agricultural techniques. However, recently developed Fe-biofortified crop varieties with enhanced Fe absorption capability provides an attractive alternative for analyzing crops for their performance under prevalent Fe deficiency in soils. In this study, contrasting wheat varieties for Fe content including WH 1105 (Non-biofortified wheat) and WB-02 (Biofortified wheat) were used for evaluating their growth performance under varied Fe supply as 100 μM (Control), 50 μM (T1), 10 μM (T2) and 0 μM (T3) under controlled growth conditions using hydroponic system. Effect of Fe deficiency as compared to Fe sufficiency was studied in plant samples at two stages of growth after treatments i.e., Stage I at 14-16 days and Stage II at 28-30 days, with analysis of various morpho-physiological (root length, shoot length, chlorophyll content and leaf area) and biochemical parameters (SOD and catalase activity, total antioxidant activity, and content of MDA, H2O2, proline, phytosiderophores and Fe). It was observed that with increased Fe deficiency under treatments T2 and T3, shoot length decreased by 23.8% and 43.6% in Stage I and 4.4% and 27.1% in Stage II, respectively, for genotype WH 1105. Comparable to this, WB- 02 also follows similar trend. A significant decreased Chlorophyll content (SPAD) in WH 1105, the chlorophyll content decrease by 10% to 67% at Stage I, and 16% to 84% at Stage II. In WB-02 increase in chorophyll from 18.9% to 66.3% was observed under increased Fe- deficiency. Variety WB-02 performed better that WH 1105 in terms of chlorophyll content under given Fe treatments as compared to control. An increase in Malondialdehyde (MDA) content (7.3% and 25.6% in WH1105 at Stage I, and 15.9% and 38.1% at stage II where as in WB-02, the content increased by 5.24% and 14.59% and by 18.3% and 27.8% Stage I and II respectively and H2O2 content 4.24% and 7.75% in the Stage I and 4.39% and 11.72% in Stage II in WH 1105, was observed, and in WB-02, it increased by 5.53% to 9.28% in Stage I, and in the Stage II, it increased by 5.87% and 11.08% with increase in Fe deficiency. Higher content of MDA and H2O2 were noticed from wheat variety WH 1105. The activities of antioxidant enzymes including superoxide dismutase (SOD) and catalase (CAT) was increased with increase in Fe-deficiency at both growth stages. Higher SOD activity was noticed from wheat variety WB-02 (in roots), WH 1105 (in shoots). Higher CAT activity was noticed from wheat variety WB-02 (in roots and shoots). The increased phytosiderophores content was observed with increase in Fe-deficiency in plants, with more PS content from wheat variety WH 1105. The Fe content decreased with increased Fe deficiency with comparatively higher uptake of Fe in roots by wheat variety WH 1105 and shoot by wheat variety WB-02..Based on the outcome of study, it might be concluded that the contrasting wheat varieties have varied biochemical and morpho- physiological responses towards Fe deficiency. Preliminary findings in this study suggested biofortified wheat variety WB-02 as better performer under given Fe deficiency treatments over WH 1105 owing to varied biochemical and morphological responses as observed. The study might have implications in development, evaluation and promotion of biofortified or Fe deficiency tolerant crops for cultivation in soils with prevailing Fe deficiency.ThesisItem Open Access Development, characterization and application of bionanoconjugate on terminal heat stress in wheat (Triticum aestivum L.)(CCSHAU, Hisar, 2023-04) Narender Mohan; Ajay PalTo ensure sustainable farming practices, nanotechnology in modern agriculture must address the worldwide issues of biotic and abiotic stress (Terminal Heat Stress) brought on by climate change and the population load on farms. Numerous biopolymers, including alginate, cellulose, chitin, and chitosan, have increased use in the new nanotechnology period to create new materials useful in crop growth and protection. Chitosan is the most extensively studied biomaterial in nanotechnology because it is the second most prevalent and conveniently accessible biopolymer after cellulose. Due to its unrestricted qualities, such as biocompatibility, antibacterial, biodegradability, and non-toxicity for plants, it is employed primarily as nanochitosan instead of as bulk throughout the world. Compared to pure/bulk chitosan or direct use of ionic metals, nanochitosan biopolymer has been deemed more valuable for its involvement in chelation with metal ions, such as Zn, Cu, Fe, etc. Herein we incorporated plant hormones (Salicylic acid) and metal (Zn) ions into ongoing cross linking of chitosan and TPP to create nano-composites/conjugates, leading to more pronounced and efficient effects on seed germination and screen house. The current study shows that the genotype, species, growth stage, ambient conditions, and ingredients/nutrients affect how plants react to BNCs. It is further inferred that applying BNCs with co-encapsulated SA and Zn improved the crop's resistance to stress. In combination with the slow-release capability of BNCs, it considerably improved the germination potential, antioxidant system, crop yield, and grain quality of both WH-1124 and WH-542, according to laboratory and pot studies. Additionally, we conclude that the synthesis of these BNCs may be accelerated further to assess their efficacy on other crops, which could be advantageous for the technical translation.ThesisItem Open Access Development, characterization and efficacy of bionanocluster of indole acetic acid and cobalt chloride on seed germination and yield in Cotton (Gossypium hirsutum L.)(CCSHAU, Hisar, 2023-09) Vikram Singh; Mandhania, ShiwaniIn the present investigation, Co2+ and indole-3-acetic acid were combined to create chitosan- Co2+-IAA BNCs. The mean hydrodynamic diameter, PDI value and zeta-potential of BNCs were recorded as 481.9 nm, 0.536 and +22.00 mV, respectively. Effect of seed priming with 100 ppm BNCs and four time foliar application of 100 ppm BNCs on seed germination, seedling growth, and boll drop was studied in H 1098i genotype of G. hirsutum L. Cotton seeds subjected to BNCs priming displayed higher percent germination and seed vigor index-I (92.14 %) compared to hydroprimed seeds due to elevated activities of glyoxylate cycle enzymes i.e. isocitrate lyase and malate synthase, which increased the conversion of fats to carbohydrates. Four times foliar application of 100 ppm BNCs increased leaf area, plant height, total chlorophyll content, photosynthetic rate, ACC synthase activity, indole-3-acetic acid content, boll number, boll weight ultimately increasing seed cotton yield by 20.37 %. However, ACC oxidase, cellulase, pectinase activities and boll drop significantly decreased compared to control. It is concluded that application of BNCs comprising of chitosan, IAA and Co2+ via seed priming and foliar mode is novel, economical, and environment friendly.ThesisItem Open Access Effect of exogenous sulphur in alleviating lead toxicity in Indian mustard [Brassica juncea (L.)](CCSHAU, Hisar, 2023) Manne Hemanth Kumar; Nisha KumariThe Indian mustard [Brassica juncea (L.)] is one of the well-known plant for phytoextraction of heavy metals. The present study was designed to elucidate whether exogenously applied sulphur (S1; 100 ppm, S2; 200 ppm sulphur) alleviate lead (Pb)-induced (Pb1; 100 ppm, Pb2; 200 ppm and Pb3; 300 ppm Pb) stress in the leaves of Brassica juncea. Lead was applied in the form of lead nitrate five days prior to sampling and sulphur applied in the form of zinc sulphate at three days before sampling and samples were collected on 30, 60 and 90 DAS. Lead stress of 100, 200 and 300 ppm has considerably reduced plant growth as evidenced by increased H2O2 contents of 303.64, 317.39 and 238.89 % at 300 ppm of 30, 60 and 90 DAS, respectively, and they further caused the peroxidation of lipid membranes. This disruption increased more leakage of electrolytes, and created ionic imbalance. Further, alteration of chlorophyll pigment by replacing its central magnesium atom has reduced photosynthetic rate by 27.24 % at Pb3 stress, in respect to control. The redox status of the cell got altered by lead toxicity, depleting the AsA and GSH pool metabolites. Besides, GR, APX and POX antioxidative enzyme activities have shown highest increase under Pb3 stress by 51.97, 57.84 and 121.45 %, at 30, 60 and 90 DAS, respectively. In contrast, S application showed an increased AsA-GSH contents by 15.30, 9.40 and 7.89 % at S1 spray and 38.12, 23.44 and 33.40 %, respectively, at 30, 60 and 90 DAS at S2 spray. Sulphur-metabolites such as ATPS, OASTL and GST activities enhanced upon stress and resulted in further augmentation upon sulphur supplementation. Sulphur application to the Pb-stressed Brassica plants has reduced the oxidative stress by decreasing EL content by 11.69 % and MDA contents by 13.79 % at 90 DAS, respectively, in respect to Pb3-stressed plants. This decrease in oxidative stress biomarkers was due to an increase in AsA-GSH thiol pool and antioxidative enzyme enhancement under 100 and 200 ppm sulphur application to the Pb-stressed plants. Besides, S2 application to the Pb3+S2 stressed plants has increased phytochelatins by 522.47, 352.60 and 443.48 %, at 30, 60 and 90 DAS, respectively, in respect to Pb3 stress and called for PC-based detoxification of Pb. Furthermore, lead stress shown a decreased relative gene expression of BjSULT1.a gene and increased BjPb2.1, BjATPS1, BjOASTL2, BjABCC2 and BjGR1 gene expression. Moreover, S2 spray increased gene expression levels by 1.32, 1.49, 2.42 and 2.24 folds of BjSULT1.a, ABCC2, BjATPS1 and BjOASTL2, respectively, under Pb3 stress. Sulphur spray of S1 and S2 to the Pb-stressed Brassica juncea plants increased morpho-physiological parameters and yield and yield parameters. In the view of present findings, greater decrease of oxidative stress (H2O2, MDA and EL), increased enzymatic, non-enzymatic antioxidants, phytochelatins, enhanced relative gene expression levels and maximum seed yield was found under exogenous supplementation of 200 ppm sulphur and has ameliorated lead stress.ThesisItem Open Access Physiological and biochemical changes in response to iron deficiency in pearl millet(CCSHAU, Hisar, 2022-08) Ritu; Dr. Vinod KumarIron (Fe) is the most essential micronutrient required for plant growth and development, being required as a redox active metal involved in various physiological and metabolic processes. About 1/3rd of world soil is deficient in Fe affecting plant growth and development leading to poor yield as well as nutritional quality. It is difficult to manage Fe deficiency to plants through agronomic practices. However, recently developed Fe biofortified crop varieties with higher Fe uptake potential provides an attractive alternate for evaluation for their performance under prevailing Fe deficiency in soils. In this study, Biofortified pearl millet hybrid HHB-299 developed by CCSHAU, Hisar was used along with popular non-biofortified hybrid HHB-67(I) for evaluating these contrasting Fe containing genotypes for their growth performance under varied Fe supply (100, 50, 10 and 0 µM Fe in nutrient solution, respectively) under controlled conditions. Effect of Fe deficiency as compared to Fe sufficiency was studied at 5-leaf and panicle initiation stages of pearl millet genotypes with analysis of various morpho-physiological (root length, shoot length, chlorophyll content and leaf area) and biochemical parameters (SOD and catalase activity, total antioxidant activity, and content of MDA, H2O2, proline, phytosiderophores and Fe). It was observed that with increase in Fe deficiency under treatments T1 to T3, shoot length was decreased in the range of 0.92- 38.9% and 9.5-41.7% at 5-leaf and panicle initiation stage, respectively, for genotype HHB-67(I), as compared to control. In comparison, significantly, less decrease in shoot length was observed for biofortified genotype HHB-299. Similarly, increased rate of chlorosis was observed in both the genotypes, with increased Fe deficiency. Overall analysis of data for antioxidant activity revealed its values in the range of 18-76% at 5-leaf stage and 32.3- 56.1% at panicle initiation stage, respectively, for both the genotypes. With increase in Fe deficiency, the phytosiderophores content was noticed an increase in the range of 55.5-69.2% and 53.3-68.1% at 5-leaf and panicle initiation stages of genotype HHB-67(I), respectively. In the genotype HHB-299, it was increased up to 81.7% and 75% at 5-leaf and panicle initiation stage under increased Fe deficiency treatments. The Fe content in shoots and roots was reportedly decreased under Fe deficient conditions, for both the stages and genotypes. Based on the outcome of study, it is concluded that the contrasting pearl millet genotypes have varied biochemical and morpho-physiological responses towards Fe deficiency. Preliminary findings in this study suggested HHB-299 as better performer under given Fe deficiency treatments over HHB-67(I) owing to its less affected biochemical and morphological responses as observed. The study might have implications in development, evaluation and promotion of biofortified or Fe deficiency tolerant crops for cultivation in soils with prevailing Fe deficiency.ThesisItem Open Access Biochemical and protein profiling in response to application of herbal nano-formulation(s) on shelf life of guava (Psidium guajava L.)(CCSHAU Hisar, 2022-09) Anju Rani; Tokas, JayantiThe guava (Psidium guajava L) is a resilient, easily adaptable and evergreen tree of the Myrtaceae family. Guavas are nutrient rich fruit having high content of vitamin A, vitamin C, antioxidants and minerals but the post-harvest losses stand at 20-40% of the produce. Herbal NFs were used to prevent post-harvest losses. Initially, different herbal extracts of giloy (leaves and stem) and ajwain seeds were prepared using different solvents and extraction procedure. Ajwain methanol (E2), ajwain methanol-acetone (7:3) (E3), giloy methanol (E16) and giloy acetone (E17) leaves extracts were selected for NFs synthesis. Three NFs viz. sodium alginate: CaCl2 (T1), sodium alginate: CaCl2: ajwain extract (T2), and sodium alginate: CaCl2: giloy extract (T3) were selected for further treatment. Physiological, quality, minerals, cell wall and ripening related parameters were analyzed every alternate day upto 10 DAT during storage at 25±2 °C and 32±2 °C. The results revealed that physiological loss in weight increased and fruit firmness decreased during storage. The quality parameters viz., total sugars, reducing sugars total soluble solids and ascorbate increased initially and then decreased at both the storage temperatures. The percent increase in total soluble sugars, reducing sugars and TSS on 10th day was maximum in control (34.8 and 36.6%; 70.7 and 76.3%; 31.8 and 34.8%) followed by T1 (29.3 and 30.0%; 64.6 and 66.2%; 15.7 and 25.1%), T3 (17.3 and 23.1%;42.5 and 49.4%; 5.1 and 9.5%), and T2 (14.9 and 20.7%; 36.4 and 44.6%; 3.7 and 8.9%) NFs at 25±2 °C and 32±2 °C. Whereas, Titratable acidity, crude proteins and antioxidant activity reduced during storage. Minerals content decreased during ripening in all the treatments and minimum decrease was recorded in T2 NFs treated fruits. Activity of cell wall degrading enzymes cellulase and polygalaturonase increased as the storage period increased whereas pectin methyl esterase activity increased initially and then decreased. The maximum percent increase of cellulase and PG was observed in control (272.6 and 365.3%; 285.3 and 325.3%) fruits followed by T1 (225.6 and 343.6%; 271.0 and 306.5%), T3 (211.2 and 318.7%; 224.1 and 272.2%) and T2 (201.3 and 292.9%; 215.2 and 266.5%) NFs treated fruits at 25±2 °C and 32±2 °C. Ethylene peak was observed on 4th and 6th day of treatment at 25±2 °C and 32±2 °C and similar trend was observed for ACC oxidase activity. The results revealed that Sodium alginate: CaCl2: Ajwain (T2) NF was most effective in controlling post-harvest losses and increased shelf life by upto 10-12 days. Lesser quality degradation took place at 25±2 °C as compared to 32±2 °C. The, control and T2 NF treated fruits were further analyzed for proteomics study using 2D-Electrophoresis and LC-MS/MS. Maximum proteins concentrated at higher pH and more in the range of 20-120 kD in 2D gel. Analysis of LC-MS/MS results using MASCOT software revealed more than 100 differentially expressed proteins (DEPs), out of which 35 proteins were up-regulated and 20 proteins were down-regulated having fold change ≥1.5 and ≤0.12, respectively, which were further analysis using GO software. Proteins were classified on the basis of molecular, biological and cellular functions. Protein profiling elucidated the expression of some novel proteins which might play a role in delayed ripening.ThesisItem Open Access Development of herbal nano-formulations for enhancing shelf life of guava (Psidium guajava L.) fruit(Chaudhary Charan Singh Haryana Agricultural University hisar, 2022-09) Gupta, Pernika; Tokas, JayantiGuava (Psidium guajava L.), member of the family Myrtaceae is an important fruit of tropical and subtropical area of world. It is also known as ―apple of the tropics‖. Nutritionally, guava fruit is a rich source of ascorbic acid but the post-harvest losses stand at 20-40% of the produce. Herbal extract based edible nano-formualtions were used to prevent post-harvest losses. Initially, different herbal extracts of ashwagandha and menthe leaves were prepared using different solvents. Three nanoformulations viz. Alg-Lys (T1), Alg-Lys:menthe extract (methanol) (T2 ), and Alg- Lys:ashwagandha extract (methanol) (T3) were selected for further treatments. Physiological, quality, cell wall components & degrading enzymes, antioxidative and ripening related parameters were analyzed every 3 day upto 12 DAT during storage at turning and mature green stage of Hisar Safeda and Hisar Surkha. The results revealed that physiological loss in weight increased and fruit firmness decreased during storage. The quality parameters viz., total soluble sugars and total soluble solids; antioxidative enzymes viz., superoxide dismutase, catalase, ascorbate peroxidase activity and ascorbate content increased initially and then decreased in both Hisar Safeda and Hisar Surkha. Overall decrease in total soluble solids was delayed in T3 treated guava fruits which was significantly higher than in control fruits in Hisar Safeda and Hisar Surkha. Whereas, titratable acidity, crude protein content, glutathione reductase and peroxidase activity reduced during storage. Activity of cell wall degrading enzymes cellulase and polygalaturonase increased as the storage period increased whereas pectin methyl esterase activity increased initially and then decreased. The amount of ethylene evolution and activity of ACC Oxidase in two cultivars of guava fruits increased initially and during further ripening process, the activity reduced. The results revealed that Alg-Lys:ashwagandha (T3) nanoformulation was most effective in controlling post-harvest losses and increased shelf life of Hisar Safeda upto 5 days and Hisar Surkha upto 3 days. The, control and T3 nanoformulation treated fruits were further analyzed 2D-Electrophoresis and LC-MS/MS and it was observed that maximum proteins had concentrated at higher pH and more in the range of 20-120 kD. Analysis of LC-MS/MS results using MASCOT software revealed more than 60 differentially expressed proteins (DEPs), which were involved in energy and carbohydrate metabolism, stress response, defense, biosynthesis, hydrolase, secondary metabolism, amino acid and nucleic acid, protein synthesis and biogenesis of cellular components having fold change ≥1.5 which were considered for further analysis. Using GO classification, major pathways involved in molecular, biological and cellular functions in Hisar Safeda fruit ripening were classified. Proteins were classified on the basis of molecular, biological and cellular functions. Protein profiling elucidated the expression of six novel proteins which might play a role in delayed ripening.ThesisItem Open Access Immobilization of xylanase from Bacillus pumilus over aluminum oxide beads(CCSHAU, Hisar, 2021-09) Tanwar, Ekta; Nagar, SushilIn present investigation the xylanase from Bacillus pumilus SV-85S was covalently immobilized over aluminum oxide beads. The immobilization yield and efficiency were 48.05 and 83.13%, respectively. Some surface modifications were observed through SEM analysis on xylanase bound aluminum oxide beads. The bands at 3440 and 1635 cm-1 were observed through FTIR analysis over aluminum oxide beads, glutaraldehyde treated beads and immobilized xylanase. Optimum pH and temperature for free and immobilized enzyme were 7.0 and 55ᵒC, respectively. Increment of Km and Vmax values was observed in immobilized enzyme by around 28.96 and 34.61 % respectively. The pH, thermal and storage stability were improved after immobilization. The Immobilized xylanase retained 65 % of its initial activity even after 5 cycles of reusability. The increase in Half-life (t1/2) and D- value whereas decrease in kd value after immobilization, indicates that thermal stability of xylanase was enhanced. Thermodynamic parameters (ΔH, ΔG and ΔS) were enhanced after immobilization of xylanase. The decrease in activation energy in case of immobilized xylanase indicates that catalytic efficiency was enhanced. Hence, the present investigation concludes that immobilization of enzyme is important due to its considerable reusability and stability, and advantageous for industrial application.ThesisItem Open Access Comparative studies of free and immobilized Aspergillus phytase(CCSHAU, Hisar, 2021-09) Mallesh, Gampa; Nagar, SushilIn this investigation the phytase from Aspergillus oryzae SBS50 was immobilized over activated aluminum oxide beads. The immobilization yield and efficiency were 47.4 and 71.2 %, respectively. Morphology and functional groups of activated beads and immobilized phytase were determined through SEM and FTIR analysis. The immobilized phytase retained 51.5 % of its initial activity even after 5 cycles of reusability. The optimum pH and temperature were shifted from 5.5 to 4.0 and 50 to 60 C after immobilization. The Km value was 2.61 and 3.09 mM, whereas as Vmax value was 32.25 and 43.29 IU/ml for free and immobilized phytase, respectively. The pH stability, thermal stability, storage stability and thermodynamic parameters (H, G, and S) were enhanced after immobilization The increase in Half-life (t1/2) and D-value whereas decrease in Kd value after immobilization indicates that thermal stability of phytase was enhanced. Thus, the investigation concludes that immobilization of phytase causes in improvement of many properties, so it is highly beneficial to immobilize the phytase.
