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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 Composting of paddy straw using additives along with microbial consortia(CCSHAU, Hisar, 2022-07) Vijay Kumar; Saharan, Baljeet SinghIn the present investigation, about 26 isolates (10 bacterial and 16 fungal isolates) available in Soil Microbiology laboratory, Department of Microbiology CCS HAU Hisar, were screened for enzyme activities including cellulase, xylanase, laccase and biosurfactant production. In case of bacterial isolates, maximum cellulose hydrolysis index (5.33) was exhibited by RC13.1 and RC3.5 isolate on CMC agar plate. Isolate RC13.1 has also shown maximum xylan hydrolysis index (4.43) laccase and biosurfactant production. In case of fungal isolates, (maximum cellulase (3.28), xylanase (2.32), laccase (qualitatively) was exhibited by H2, H3 and H16 isolates, respectively. H16 isolate also exhibited maximum biosurfactant production confirmed by oil displacement. The bacterial isolate, M2 showed maximum CMCase (0.33 IU/ml), Fpase activity (0.52 IU/ml) activity and second highest xylanase (7.63 IU/ml) and followed by laccase activity (5.2 IU/ml). In case of fungal isolates, maximum CMCase (0.31 IU/ml), and Fpase activity (0.42 IU/ml) were exhibited by H2 isolate. While isolates H3 and H16 showed maximum xylanase (16.11 IU/ml) and laccase (8.4 IU/ml) activities. Based on these activities and compatibility to each other, two bacterial (M2 and RC13.1) and three fungal (H2, H3 and H16) isolates were used for microbial consortium. Microbial consortium along with additives including FYM, cattle dung, poultry manure and mustard cake was analyzed for paddy straw degradation in flasks, trays, and pits. Treatment T5 having paddy straw + microbial consortium + poultry manure was recorded with lowest total organic carbon and maximum potassium content. Maximum total nitrogen and phosphorus content was observed in treatment T6 having paddy straw + microbial consortium along mustard cake after 90 days of composting. All the treatments having additives along with microbial consortium, had significantly higher microbial activity (dehydrogenase), humic substances and better pH, EC and germination index. On the basis of selected morphological and biochemical characters, the isolates RC13.1 and M2 were identified as Pseudomonas sp. and Bacillus sp., respectively. Based on morphological characteristics, fungal isolates H2, H3, H16 were identified as Aspergillus sp., Aspergillus sp. and Phanerochaete sp., respectively.ThesisItem Open Access Effect of paddy straw compost on microbial communities and plant growth of Mungbean (Vigna radiata L.)(CCSHAU, Hisar, 2023-07) Rekha; Malik, KamlaWorldwide, agricultural residue burning has become a serious environmental issue that has a very bad effect on human and animal health. Burning of agri-residues not only disturbs the soil structure but huge loss of plant nutrients, effect microflora and causes greenhouse gas emissions. Paddy straw and sugarcane bagasse are agricultural waste which composed of lignocellulose that cannot be degraded easily (Rath et al., 2022). Their direct use causes immobilization of nutrients, so composting arises as a safe option, resulting in the reusability of nutrients contained in these residues (Malik et al., 2021). The present study was investigated on effect of paddy straw compost on microbial communities and plant growth of Mungbean (Vigna radiata L.). Bacteria were maximum in the mesophilic phase and the fungi at thermophilic phase, while actinomycetes in the maturation phase in treatment (T3-Paddy straw (4) + Poultry waste (1) + MC). Alkaline phosphatase and cellulase were maximum in T3 at 30 days of composting while dehydrogenase (DHA) activity was maximum at 60 days in T5. The temperature of compostable material increased from 38°C to 46°C after 40 to 45 days of composting and then decreased. Total organic carbon of the compostable material in different treatments decreased more in T3 followed by T4 due to losses of C and total N increased due to accumulation of nitrogen up to 90 days. The quality of compost was tested by as C:N ratio, NPK content, humic substances and germination index. Total nitrogen was increased from 0.56 to 1.44%, similar pattern was also observed in total phosphorous and potassium followed. Maximum reduction in C/N ratio was observed in T3 (22.29%). The C/N ratio of compostable materials ranged from 85.50 initially and dropped down to 22.29 after 90 days of composting. The amount of humic substances was highest in treatment T3 compared to all other treatments. Among all treatments, the germination index was highest in T3 followed by T4. The T3 compost was selected for growing Mungbean (Vigna radiata L.) variety MH-421 in pot house conditions. Among the different treatments, T3 (60% soil + 40% PSW + PW + MC) showed highest microbial count, plant height (35.59 cm and 49.10 cm at 30 days and 65 days after sowing respectively), plant biomass, more nodule number, highest crude protein content of grains and seed yield as compared to other treatments.ThesisItem Open Access Covalent immobilization of bacterial alkaline protease and its application in poultry feed(CCSHAU, Hisar, 2023-06) Kamaldeep Kaur; Sindhu, MeenaProteases are enzymes that hydrolyse peptide bonds in proteins. Proteases represent 60% of the enzyme market and around 66% of the proteases are derived from the microbial sources (Hadedy et al., 2023). Microorganisms act as an important source of enzymes because of their vast distribution, high reproduction, and potential to be genetically altered for higher productivity. Both bacteria (Bacillus subtilis, Bacillus licheniformis etc.) and fungi (Aspergillus oryzae, Aspergillus niger etc.) are capable of producing large amounts of protease enzymes. In industries, enzymes lack reusability during hydrolytic process, which influences overall economy of the process. This problem can be addressed by immobilization of enzyme on inert material (Wu et al., 2020). Immobilization convert enzyme from soluble form to insoluble form thus enhances stability and reusability. The immobilized enzyme has many advantages over the use of free enzymes such as: increased functional efficiency, reusability, increased product stability etc. The hydrolysis of poultry feed with protease aids in the digestion of dietary proteins in the gastrointestinal system by breaking down ingested proteins into their constituent amino acids for absorption.In the present study, protease enzyme was produced from bacterial isolate KTP9 under submerged fermentation having activity 17.43 IU/mL unit and 3.626 mg/ml of protein content under submerged fermentation. Ammonium sulphate precipitation increased the specific activity from 4.80 to 22.45 IU/ ml with 4.67 purification fold. Bacterial KTP9 free protease have optimum pH 8.0, temperature 35°C and found stable upto 28 days on storage at 4°C. The covalent immobilization of protease over aluminium oxide beads was performed. The immobilization yield and immobilization efficiency were found as 40.54% and 89.72%, respectively. Optimum conditions of immobilization were 1% Glutaraldehyde as linker with 1h and 1.5h cross-linking and coupling time, respectively. Upon immobilization, shift in temperature (35°C to 45°C) and pH optima (8.0 to 10.0) was observed as compared to free protease. The immobilized protease was found to retain 50% residual activity upto 35 days of storage at 4°C. Enzyme after immobilization can be reused upto 6 cycles with 50% residual activity. The immobilized protease showed 76% protein hydrolysis in 120 minutes as compare to bacterial KTP9 free protease.ThesisItem Open Access Isolation and characterization of plant growth promoting salt tolerant bacteria from Suaeda fruticosa L. rhizosphere(CCSHAU, Hisar, 2023-06) Somu Nagajyothi; Kayasth, MonikaSoil salinity is one of the most severe environmental factors that limits the productivity of crops in arid or semi-arid regions around the world. The present investigation was undertaken to isolate and characterize plant growth promoting salt-tolerant bacteria. Rhizospheric soil samples were collected from Suaeda fruticosa halophyte in Hisar and were screened for plant growth promoting salt tolerant bacteria. Eighteen isolates were retrieved and analyzed for salt tolerance at various NaCl concentrations (2% to 10% NaCl). Twelve isolates were found salt tolerant at varied salt concentrations and evaluated for the plant growth promoting traits. All the salt tolerant isolates showed IAA production with maximum in isolates SA3, SR1, SR2, and SS1. Ammonia excretion was maximum in isolate SR1 and minimum in isolate SR5. Ten isolates showed phosphate solubilization and eight isolates showed Zn solubilization. All twelve salt tolerant bacterial isolates were found negative for siderophore production. Four isolates SA3, SR1, SS1 and SR2 with significant salt tolerance and PGP traits were characterized biochemically. Based on biochemical results, the tentative genus identified as Bacillus (SA3, SR1, and SR2) and Serratia (SS1). The effect of salt-tolerant PGP bacterial isolates on seed germination of wheat, pearl millet, mung bean, and chickpea under different salt concentrations was determined. By increasing the salt concentration, salt tolerant PGP isolates showed better germination than the control. However, isolates SA3 and SR1 showed maximum seed germination and were further identified by 16s rDNA gene sequencing. 16S rDNA gene sequencing of isolate SA3 showed 99.64% similarity with Bacillus safensis and SR1 isolate showed 99.78% similarity with Bacillus cereus.ThesisItem Open Access Impact of co- inoculation of rhizobium and trichoderma on nodulation efficiency, plant growth parameters and biocontrol efficacy in mung bean crop (Vigna radiata L.)(CCS HAU, Hisar, 2022-08) Pallaganti Sowmya; Ajay KumarMung bean (Vigna radiate L.) belongs to the Leguminosae family and one of the most popular short duration pulse crops of India. It is one of the essential food legumes cultivated worldwide. is an excellent source of high-quality protein and contains about 22-25% protein, 1.0-1.5% oil, 3.5-4.5% fibre, 4.8% ash and 62-65% carbohydrates on dry weight basis. It is easily digestible and hence it is suitable for all age group of people and green fodder to feed animals. Rhizobia are Gram-negative bacteria that can form root nodules in leguminous plants and provides the nitrogen through fixation process which can limit the use of chemical fertilizer. Trichoderma belongs to the Deuteromycetes class of fungi and has been exploited as biocontrol agents against a diversity of plant pathogenic fungi. In the present investigation, mung bean cultures and Trichoderma isolates were screened for their compatibility. Trichoderma asperellum and Rhizobium MB 17a were found compatiblemwere then optimized for their growth interaction. The coculture was found to be more effective when tested for their antagonistic activity against Macrophomina phaseolina. The coculture recorded higher plant growth promoting traits including IAA production, ammonia excretion phosphate solubilization, , zinc solubilization, siderophore production, HCN production and ACC utilization. were evaluated for growth promotion of mung bean under pot culture conditions during 2020. Rhizobium and Trichoderma coculture gave best results in improving growth and seed yield of mung crop.ThesisItem Open Access Antioxidative and antimicrobial activity of bioactive peptide from Quinoa (Chenopodium quinoa) fermentation(CCSHAU Hisar, 2022-08) Khan, Sameer; Sindhu, MeenaQuinoa (Chenopodium quinoa Wild) also known as Inca rice is a pseudocereal having immense nutritional benefits (Dakhili et al., 2019). It is also called perfect food because of its nutritional content which contains proteins, vitamins, minerals (manganese, phosphorous, iron, zinc), dietary fibre, and the dietary minerals. It is gluten free, so can be used as food for people having gluten allergy (Galante et al., 2020). The proteins present in the quinoa can be broken down in to smaller peptide of 2-20 amino acid chain having beneficial effect on human health. Present investigation targeted bioactive peptide production by fermenting quinoa using three probiotic bacterial strains i.e. Lactobacillus plantarum, Lactobacillus rhamnosus and Lactobacillus fermentum. Submerged fermentation was performed using these probiotic bacterial cultures for different time intervals (24h, 48h and 72h). Maximum decrease in pH 6.72 to 3.72 was observed during fermentation of Quinoa with Lactobacillus fermentum. Maximum peptide concentration (13.6 mg/mL) was observed during 48 h of Quinoa by L. fermentum using OPA (o-phthaldialdehyde) spectrophotometric assay and it was selected for further analysis. SDS –PAGE segregated the peptides as per molecular weight and maximum peptides obtained were belong to size range 3-20 kDa. Peptides were separated from the supernatant of fermented quinoa using TCA (tri chloro acetic acid) precipitation and categorized on the basis of different size using membrane filter having cut off size 30kDa, 20kDa and 5kDa. The antioxidant scavenging activity were checked using DPPH (2,2-diphenyl picrylhydrazyl) radical assay (Yen and Duh 1994), ABTS (2,2 azino-bis (3 ethylbenzthiazoline-6sulphonic acid) assay (Joshi et al., 2015) and ferrous ion chelating ability (Do et al., 2014). Maximum DPPH radical scavenging ability (16 %) and maximum ABTS (26.2%) scavenging ability was observed in peptide below 5 kDa produced by Lactobacillus fermentum as compare to control. Ferrous ion chelating ability were maximum (26.83 mg AE/gmDW) in <5 kDa peptides formed by L. plantarum as compare to control (14.69 mg AE/DW). Antimicrobial activity of these different size peptide were also observed against Staphylococcus aureus, E.coli, Fusarium oxysporum and Candida sp.. Maximum zone of inhibition was observed in 5-20 kDa peptide (27mm) produced by Lactobacillus plantarum against Staphylococcus aureus followed by below 5 kDa peptides (26mm) by Lactobacillus fermentum. Maximum antimicrobial activity against E.coli (26mm) and Candida sp. (24 mm) was observed in peptides below 5 kDa produced by Lactobacillus fermentum. No antifungal activity was shown by peptides against Fusarium oxysporum. Significant antioxidant and antimicrobial activity were observed in peptides produced by submerged fermentation using Lactobacillus fermentum and Lactobacillus planatrum hence their application can be explored in fat rich food to prevent rancidity and also in post harvest disease management to enhance shelf life of perishable vegetables/fruits.ThesisItem Open Access Bioconversion of microbially hydrolysed Water hyacinth (Eichhornia crassipes) into ethanol using Saccharomyces cerevisiae HAU-1(Chaudhary Charan Singh Haryana Agricultural University hisar, 2022-12) Sonia; Mehta, ShikhaBioethanol is considered as a potential liquid fuel due to limited natural resources. Lignocellulosic biomass is investigated as a potential substrate for bioethanol production. Water hyacinth (Eichhornia crassipes) is one of the most abundant lignocellulosic materials, which consist of about 35% hemicellulose, 26% cellulose and 11 % lignin. A pre-treatment process is essential to remove lignin, reduce crystallinity and increase the porosity of material making cellulose desirable to hydrolysis. To hydrolyse the cellulose, cellulases are required which increase the cost of ethanol production. Therefore, to make the process cost-effective, there is need to utilize microbial cellulase. In the present study, a total 43 bacterial and 16 fungal isolates were retrieved from 3 different samples such as soil from CCS HAU, Hisar, decaying woody material from village Mirchpur, Hisar and cattle dung from CCS HAU, Hisar. All the bacterial and fungal isolates were screened for cellulase production on the basis of zone of hydrolysis on carboxymethyl cellulose (CMC) agar medium. Out of 43 bacterial isolates, 13 bacterial isolates showed zone formation on CMC agar medium and three isolates i.e. SB2, SB4 and SB10 showed large zone. Out of 16 fungal isolates, 10 fungi showed the zone formation on CMC agar medium and WF1 isolate showed large zone of hydrolysis. Cellulase activity of SB4 and WF1 was determined and found to be 276.83 IU/mL and 230.62 IU/mL, respectively. Bacterial isolate SB4 was biochemically characterized and stained using Gram’s staining. SB4 was found to be Gram positive. Pre-treatment of water hyacinth was done using acid (1% H2SO4) as well as alkali (2% NaOH). Acid pre-treated water hyacinth consisted of 30.23% cellulose, 23.50% hemicellulose and 5.83% lignin and alkali pre-treated water hyacinth consisted of 29.86% cellulose, 24.30% hemicellulose and 6.13% lignin. Crude enzymes of bacterial isolate SB4 and fungal isolate WF1 were used for enzymatic hydrolysis. Hydrolysis of acid pre-treated water hyacinth with crude enzyme obtained from SB4 isolate released 65.74 mg/g total reducing sugars after 120 min of incubation. Fermentation of hydrolysate was carried out using Saccharomyces cerevisiae HAU-1. Different conditions such as pitching rate, slurry concentration and nitrogen sources were optimized for maximizing ethanol production. Fermentation of hydrolysate of acid pre-treated water hyacinth supplemented with 0.15% urea resulted in production of 1.77% (v/v) ethanol after 72 h of incubation using crude enzyme obtained from SB4 isolate.ThesisItem Open Access Microbial valorization and fortification of agricultural wastes through composting process(Chaudhary Charan Singh Haryana Agricultural University, 2022-12) Amisha; Malik, KamlaWorldwide, agricultural residue burning has become a serious environmental issue that has a very bad effect on human and animal health. Burning of agri-residues not only disturbs the soil structure but causes of huge loss of plant nutrients, effects microflora and causes greenhouse gas emissions (Sarkar et al., 2020). Paddy straw, Sugarcane bagasse, and saw dust are agricultural waste and are composed of lignocellulose compounds that cannot be degraded easily (Perez et al., 2002). Their direct use causes the immobilization of nutrients, so composting arises as a safe option, resulting in the reusability of nutrients contained in these residues. So, the present investigation was planned with the objectives of managing paddy straw, sugarcane bagasse, and sawdust along with cattle dung and microbial consortia in different ratios using pits with an initial C/N ratio of these wastes (Raj and Antil, 2011). In log (CFU/ml), the treatment with paddy straw + sugarcane bagasse + sawdust + cattle dung + Microbial consortia (4:1:1:1) (T6) had the highest bacterial and fungal counts, while the treatment with sugarcane bagasse + sawdust + cattle dung + Microbial consortia (4:1:1:1) (T9) had the highest actinomycetes count. Alkaline phosphatase, cellulase, dehydrogenase (DHA), and protease were maximum at 30 days of composting in T6 followed by T9. The pH value is also neutral in the mature compost after 90 days. EC values are minimum with T9 followed by T6. Total organic carbon of the compostable material in different treatments decreased more in T6 followed by T9 due to losses of C and total N increased due to accumulation of nitrogen up to 90 days. The maximum reduction in C/N ratio was in T6 - 26.50 followed by T9. Total nitrogen (%) increased from 0.65 to 1.48. The same pattern was observed in total phosphorus, potassium in T6 was followed by T9. The temperature of compostable material increased from 300C to 55°C after 40 to 45 days of composting and then decreased. The quality of compost was tested by C: N ratio, CO2 evolution, humic substances, and germination index. The C/N ratio of compostable materials ranged from 78.20 to 80.05 initially and dropped down to 23.25 to 31.97 after 90 days of composting. The amount of humic substances was highest in treatment T6 compared to all other treatments. The germination index of wheat and mung bean at different treatments are as follows: T1-71-74%, T2-72-75%, T3-70-71%, T4-80-82, T5- 57-65, T6- 90-91, T7- 83-81, T8-83-85, T9-87-89, T10-83-85, T11-84-85 and T12-88-89% and germination index of mustard is less. Among all treatments, the germination index was highest in T6 followed by T9. These two treatments (T6 and T9) were selected for further fortification with poultry and vegetable waste. Among the fortified treatments, T6 + Poultry showed Organic carbon -30.40%, C/N ratio-18.54, humic acid of 142 mg/gm, CO2 evolution -101 mg per 100 gram of compost and germination index - 90 to 100% leachate of compost extract, which indicates it doesn’t have any phytotoxic effect on seedThesisItem Open Access Development of biocontrol agent against phytopathogenic fungi of tomato (Lycopersicon esculentum Mill.) crop.(CCSHAU, Hisar, 2020-07) Kapoor, Divya; Rakesh KumarIn the present study, seventy-eight bacterial isolates were retrieved from the tomato rhizosphere soil and screened for their antagonistic activity against Fusarium oxysporum f. sp. lycopersici, Alternaria solani and Rhizoctonia solani under in vitro conditions. Thirteen isolates, NAU5, NAU10, NAU13, NAU20, NAU21, NAU23, NAU29, NAU32, NAU36, NAA59, NAA60, KBA43 and KBA44 showed broad inhibition zones against A. solani measuring more than 20 mm. Six isolates, NAU19, NAU20, NAU23, NAU24, NAU27 and NAU30 resulted in broad inhibition zones against F. oxysporum f. sp. lycopersici measuring 10.1-20 mm. Six isolates, NAU20, NAU30, NAU35, LBV58, NAV66 and NAV73 resulted in broad inhibition zones against R. solani measuring 15.1-25 mm. Among the retrieved 78 isolates, 28% of rhizobacteria produced high amount of IAA and seven isolates, NAU4, NAU5, NAU10, NAU20, NAU28, KBA43 andKBV47 produced maximum IAA. The amount of IAA production by different isolates varied from 3.62μg/ml (NAV69) to 26.96μg/ml (NAU5). Among 78 isolates tested for chitinase assay, two isolates, NAU13 and NAV68 showed maximum activity. Out of 78 isolates, siderophore production was observed in 42% of retrieved bacteria and five isolates, NAU19, KBV45, KBV46, LBV56 and NAV75 produced maximum orange halo zone. HCN was produced by 10% of isolates and highest HCN was observed by 6 isolates, NAU4, NAU6, NAU20, NAU29, NAU41 and NAA59, qualitatively. Seven isolates, NAU5, NAU20, NAU30, NAU35, NAA59, NAV68 and NAV75 were selected and total of 27 treatments were prepared on the basis of antagonistic and plant growth promoting attributes tested in vitro for further evaluating under pot house conditions. Isolate NAU5 showed maximum shoot length, shoot weight, root weight, fruit yield and disease control (100%) against F. oxysporum f. sp. lycopesici. Isolate NAV75 showed maximum shoot length, shoot weight, root weight, fruit yield and disease control (87%) against A. solani. Isolate NAU35 showed maximum shoot length, shoot weight, root weight, fruit yield and disease control (100%) against R. solani. Irrespective of the fungi, isolate NAU5 showed biocontrol activity against all the three phytopathogens. The seven promising isolates showed the PGPR activities in the order NAV75> NAV68> NAA59> NAU5> NAU35> NAU20> NAU30 irrespective of the fungi in pot house studies. The isolates NAU5 against all the three fungi, NAU35 against R. solani and NAV75 against A. solani can be further tested on the tomato fields as potent biocontrol agents. On the basis of biochemical characterization isolate NAU5 was found to belong to Alcaligenes, NAU35 to Bacillus and NAV75 to Burkholderia genus.
