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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 Evaluation of plant growth promoting actinomycetes on chickpea (Cicer arietinum L.)(CCSHAU, 2019) Kavita Rani; Leela WatiChickpea is the second most important cold season food legume grown globally on a very wide area. However, global yields of chickpea have been relatively stagnant for the last two decades due to unpredictable climatic changes, incidence of diseases and extensive application of chemical fertilizers and pesticides. Actinomycetes are one of the major components of rhizosphere microbial population and are useful in soil nutrient cycling as well as plant growth-promotion. Since last few years, studies on beneficial traits of actinomycetes regarding plant growth promotion and biocontrol activities had opened new avenues for their applications in sustainable agriculture. In the present investigation, total 40 (AK1-AK40) actinomycete isolates were retrieved from different soil samples and chickpea nodules collected from CCS Haryana Agricultural University, Hisar farms. All the isolates were assessed for different plant growth promoting traits including IAA production, P solubilization (P-SI), Zn solubilization (Zn-SI), siderophore production, HCN production, salt tolerance, biocontrol potential against F. oxysporum, ACC utilization and ammonia excretion. Based on the results of various PGP traits, isolate AK3 with high P-SI (2.78) and Zn-SI (3.92); AK6 with high HCN production; AK11 with good P-SI (2.70) and Zn-SI (3.27) and AK34 with high IAA production (20.95μg ml-1) were evaluated for growth promotion of chickpea under pot culture conditions. Isolate AK3 found promising in terms of nodulation, growth and yield parameters under pot culture conditions was evaluated on chickpea variety HC-5 under field conditions (during 2017-2018). Coinoculation of AK3 along with Mesorhizobium 1233 was beneficial for plant growth and yield of chickpea. The promising actinomycete isolate AK3, identified as Streptomyces griseoruber on the basis of partial 16S rDNA sequencing, can be further tested on different locations for use as biofertilizer.ThesisItem Open Access Biochemical characterization and toxicity Studies of biosurfactants produced by yeast(CCSHAU, 2019) Dolly Rani; Sangwan, SeemaThe biosurfactant production potential of two yeasts, Meyerozyma guilliermondii YK 32 and M. guilliermondii YK 22 was investigated using total eight types of carbon sources followed by extraction and characterization of final product. Maximum biosurfactant production monitored in terms of oil displacement (9.0cm) was obtained using a mixture of olive oil and whey at 4%(v/v) concentration of each in case of M. guilliermondii YK 32. Its cell free supernatant yielded maximum 12.77g/L crude biosurfactant using acetone precipitation method. Total eight types of crude biosurfactants obtained using different carbon sources (BS1-BS8) were subjected to further characterization. The preliminary characterization using TLC revealed yellow spots in case of BS1, BS4, BS5 and BS8 having Rf 0.33, 0.41, 0.45 and 0.38, respectively indicating the presence of carbohydrate and lipids while a ninhydrin spray detected purple to reddish purple spots in BS2, BS3, BS6 and BS7 with Rf value varied from 0.23 to 0.93 depicting protein and lipid contents. The biosurfactant produced using a mixture of olive oil and whey (BS6) as substrate contained highest protein content (28.61 mg/g) followed by BS2 (20.90 mg/g) and BS1 (15.94 mg/g) while a mixture of molasses and whey supported the production of crude biosurfactant (BS8) containing highest carbohydrate (28.82 mg/g) as well as lipid content (A260nm, 16.85). All the crude biosurfactants exhibited neutral behavior during ionic charge determination. Contrary to chemical surfactant (1%SDS) which inhibited the germination of chickpea, the increasing concentration of biosurfactants supported the germination of chickpea giving a maximum germination index equaled to 423 % at 0.8% concentration of biosurfactants BS4.Further, all the biosurfactants were non toxic to yeast multiplication and did not show any antimicrobial activity against Rhizobium, a bioinoculant for nitrogen fixation. Being supportive to chickpea germination and nontoxic to yeast, the neutral biosurfactant produced by M. guilliermondii YK 32 may be tested for application in agricultural practices, cosmetics and pharmaceutical industries.ThesisItem Open Access Bioremediation of phenolic compounds in sewage water using bacterial consortia(CCSHAU, 2019) Diksha; Rakesh KumarSewage water is a rich source of organic waste and nutrients in the form of nitrates and phosphates, due to which it can be used in agriculture after proper treatment. Sewage water contains various harmful compounds like phenolics which need to be treated before discharge. The direct discharge of sewage water into natural water sources leads to the obstructive impacts on the water sources and aquatic life, sewage water becomes an open source of harmful microbes and pollutants. To demolish these problems the treatment of domestic waste water is must and its reuse for the domestic as well as agriculture purpose can become an eco-friendly solution for the conservation of water. A total of 29 bacterial isolates were retrieved from seven sewage water samples collected from Sewage Treatment Plant, CCS HAU, Hisar. All bacterial isolates were screened for phenol degradation in mineral salt medium supplemented with 250, 500 and 750 ppm concentration of phenol. Out of 29, only 8 bacterial isolates were able to grow in mineral salt medium having 750 ppm concentration of phenol. These eight bacterial isolates were further observed for growth upto 1500 ppm concentration of phenol, out of eight, only four bacterial isolates (NI6, NI5, NR1 and JC1) were able to grow upto 1250 ppm concentration of phenol. Cultural conditions, inoculum size and aeration were optimised in culture medium as well as sewage sludge. Fifteen percent inoculum and 2 LPM aeration rate were found to be optimum for efficient phenol degradation. Least residual phenol in culture medium and sewage sludge at 15% inoculum was left with bacterial isolate NI6 (265.2 ppm and 607.30 ppm respectively). Same was observed in culture medium and sewage sludge at 15% inoculum and 2 LPM aeration in lab scale fermenter for bacterial isolate NI6 (352.3 ppm and 761.27 ppm respectively). A consortium of four bacterial isolates (25% each of NI6, NI5, NR1 and JC1) was found to leave least 276.93 ppm residual phenol in sewage sludge as compared to individual isolates. Consortium was further tested for removal of phenolic compounds, COD, BOD and Heavy metal at STP, CCS HAU, Hisar. At 9th day of application, highest phenolic compounds degradation was observed in sewage sludge (120 ppm) and outlet of clarifier (172.9 ppm), thereafter phenolic compounds concentration started to increase. At 9th day COD reduced from 460 mg/L to 60 mg/L and 100 mg/L to 40 mg/L while BOD decreased from 153 mg/L to 18 mg/L and 28 mg/L to 17 mg/L in sewage sludge and outlet of clarifier respectively. Removal of Zinc and Chromium was observed in outlet of clarifier and sewage sludge.ThesisItem Open Access Characterization of thermotolerant rhizobacteria to control root rot disease in clusterbean (Cyamopsis tetragonoloba L.)(CCSHAU, 2019) Sachin; Pathak, D.V.Cluster bean [Cyamopsis tetragonoloba (L.) Taub.] is an under exploited leguminous crop which is commonly known as guar, chavli kayi, guari, khutti etc. It is grown in India, especially in South Haryana, Rajasthan, Andhra Pradesh and Maharastra in the summer season. India contributes around 80% of cluster bean to the world. Rhizoctonia solani Kuhn, a basidomycetes fungus, is the main causal organism of this disease. The control of fungus is difficult because of wide range of its host, large sclerotia are insensitive to chemical fungicides. A total of fifty eight bacterial isolates were retrieved from rhizosphere of mainly two crops (Wheat and congress grass). All the bacterial isolates were screened for Gram’s staining, spore staining, IAA production, NH3 excretion, growth on chitin as carbon source, antifungal activity against R. solani, Germination index, pre emergence mortality and Relative root elongation index (RREI). On the basis of Gram’s staining 45 isolates were found to be Gram’s +ve, remaining were Gram nagetive. Endospore staining showed 45 isolates as spore formers. A total of 56 isolates were capable of IAA production and maximum IAA producer was NAA4 (25.03 μg ml-1). Out of 58 isolates 53 isolates were capable of ammonia excretion and maximum NH3+ excretion was 4.16 μg ml- 1 by isolate no. KMI1. Only five isolates (KMI1, NAB1, NAE2, NAD1 and NAC2) were showed chitinase activity using chitin as carbon source. Ten bacterial isolates were showed antifungal activity against R. solani under in vitro conditions. Impact of 20 best cultures selected on the basis of biochemical characterization, was studied for germination in cluster bean under in vitro conditions. The maximum root length was observed with isolate NAG6 (7.86 cm) and maximum shoot length was 8.82 cm with NAG6. Maximum RREI % was observed with NAG6 which was 242.59 %. Minimum pre emergence mortality was observed with KME2. It was only 3 %. Germination index varied from 50 to 90 % and maximum GI was 90% in five cultures KMB7, KMH3, KML2, NAB1, NAD1. The best five cultures on the basis of antifungal activity, chitinase activity and other biochemical characterization were selected to evaluate under pot house conditions. Maximum root length was observed with NAE2 along with RDF (18.11cm) and shoot length was observed with NAE2 along with RDF (20.76 cm). Maximum GI was observed in case of NAE2 along with RDF (82.96 %). Minimum pre emergence mortality % observed was 8.00 % in NAE2 along with RDF. Minimum post emergence mortality % observed was 9.04 % in NAE2 along with RDF. The maximum dry shoot weight was observed in case of NAE2 along with RDF (5.075 mg). The maximum dry root weight was 0.403 mg in NAE2 along with RDF followed by 0.392 mg in NAD1 along with RDF. The maximum N uptake was observed with NAE2 along with RDF (64.75 mg/pl.). NAE2 culture performed better for root rot control and most of plant growth parameter under pot house conditions. This culture was identified as Bacillus subtilis subtilis using 16sRNA RFLP.ThesisItem Open Access Effect of conservational practices on physico-chemical and microbiological properties of soil under different cropping systems(CCSHAU, 2019) Yadav, Dhinu; Leela WatiTillage is the mechanical disturbance of soil through plowing, cultivation or digging and has been used by the farmers since ancient time that influence physical, chemical and biological properties of soil. Conventional tillage practices may adversely affect long-term soil productivity due to erosion and loss of organic matter in soils. Conservation tillage is the practice in which at least 30% of crop residues are left in the field during sowing, thus reduce soil erosion. The advantages of conservation tillage practices over conventional tillage include reducing cultivation cost, and building up soil organic matter. Conventional tillage can lead to soil microbial communities dominated by aerobic microorganisms, while conservation tillage practices increase microbial population and activity as well as microbial biomass. Long-term no tillage application increases organic carbon content, positively affecting not only soil structure, but also microbiota activity. The effect of tillage on soil microbial population has generally been studied by comparing microbial numbers, soil microbial community enzyme activities (dehydrogenase, phophatase, cellulase and urease) and microbial biomass. Enzymes are responsible for carrying out mineralization of different soil nutrients such as phosphatase for P, urease for N and dehydrogenase represents total microbial activity of soil. In the present study, soil samples (0-15 and 15-30 cm depths) were collected after harvesting of wheat from different fields under different crop rotations and analyzed for various physico-chemical and microbiological properties. The long- term zero tillage in different cropping systems affected the soil physical properties such as bulk density at 0-15 and 15-30 cm depths in the different textured soils and it was comparatively higher at 0-15 cm depth as compared to 15-30 cm depth under zero-tillage and vice-versa under conventional tillage. Chemical properties of soil such as EC, pH were not significantly affected at different depths under zero and conventional tillage but CaCO3 content of different soil samples was affected by different tillage practices at different locations to different extent. Total N, P and K and available N, P, K and S and ammonical-N, nitrate- N contents were higher at surface layer under zero-tillage. Various microbiological properties like microbial biomass carbon and nitrogen, enzymatic activities i.e. dehydrogenase, alkaline phosphatase, cellulase and urease and viable counts of bacteria, fungi, actinomycetes, Azotobacter and P solubilising bacteria were affected by tillage system under different cropping patterns. The C and N mineralization rate was found more pronounced at 0-15 cm depth but variable at different locations. Functional microbial diversity at different locations was changing with passage of time, indicating that microbial diversity changed with adoption of no-tillage practices.ThesisItem Open Access Green synthesis and characterization of plant derived nanoparticles for enhancement of biogas production(CCSHAU, 2019) Dilbag; Malik, KamlaEnergy crisis and environmental pollution are serious problems of sustainable development of the world. The problem is far more acute in the perspective of developing economies in countries like India where there is significant pressure on the available natural sources of energy. One of effective approaches to solve these energy problems is developing biomass energy such as biogas and biofuel. Biogas production is a well-established conversion technology to obtain energy from biomass that is alternative to fossil fuel. Nanotechnology can pave the way for betterment of biogas production through the use of nanocatalysts which ensure more efficient bioconversion strategies, better breakdown of substrates and more optimized output delivery. Generally, nanoparticles are prepared by a variety of chemical and physical methods which are quite expensive and potentially hazardous to the environment which involve use of toxic and perilous chemicals that are responsible for various biological risks. This may be the reason which leads to the biosynthesis of nanoparticles via green route that does not employ toxic chemicals and hence proving to become a growing interest to develop environment friendly processes. Therefore, in the present investigation was carried out green synthesized plant derived iron nanoparticles used for enhancement of biogas production from cattle dung. The nanoparticles were synthesized from neem (Azadirachta indica) and curry (Murraya koenigii) leaves with various conc. (30-70 ml) supplemented with cattle dung for biogas production. Characterization of the synthesized iron nanoparticles was done by UV-visible spectrophotometer, PSA, zeta potential analyzer (ZPA) and transmission electron microscopy (TEM). The highest absorption peak was observed at 432 nm.TEM image reveals that the FeNPs were in smaller size and spherical in shape with a diameter of 72.5 nm (NE) and 25 nm (CE), respectively. The maximum degradation (43.1%) of total solids was observed in T-5. Similar trend was observed in VS degradation and maximum degradation (15.7%) was observed in T-5. Biogas production from FeNPs (NE & CE) T-5 after 8 weeks of digestion was 104.3 litres and in T-9, it is found to be 98.6 litres as compared to control in which biogas production was found to be 85.2 litres. There was 22.4% increase in biogas production as compared to control after 8 weeks of batch anaerobic digestion. Cumulative biogas production with the addition of FeNPs attained the highest biogas yield (2537 l) in J2 as compared to J1 (2201 l) and methane contents were 77.8% as compared to control 72.3% after 8 week of digestion.ThesisItem Open Access Growth stimulation of pigeon pea (Cajanus cajan L.) by using antagonistic rhizospheric bacteria(CCSHAU, 2019) Sharma, Ruchi; Sindhu, S.S.Pigeon pea (Cajanus cajan L.) is one of the important grain legume crop in the developing countries of the tropical and subtropical regions of the world. It is an excellent source of protein (20-22%), supplementing energy rich cereal diets in a mainly vegetarian population. One of the major factors adversely affecting pigeon pea productivity is poor nodulation in fields due to prevalence of poor nodulating native rhizobia. Moreover, the crop productivity of pigeon pea is also adversely affected by biotic stresses such as Fusarium wilt (FW), Alternaria blight and Rhizoctonia root rot disease. In the present study, 83 rhizobacterial isolates and 24 rhizobial isolates were isolated from different soil samples. Screening of the isolates for antagonistic activity against three Alternaria tenuissima, Fusarium udum and Rhizoctonia bataticola showed that only 21% of the rhizobacterial isolates inhibited the growth of all the three pathogenic fungi. Four isolates i.e., HPR6, HPR22, HPR71 and HPR78 showed more than 4.0 halo zone to growth ratio against Alternaria tenuissima wheareas three isolates i.e., HPR37, HPR41 and HPR42 showed more than 4.0 halo zone to growth ratio against Fusarium udum and 3 isolates i.e., HPR5, HPR71 and HPR78 showed more than 4.0 halo zone to growth ratio against Rhizoctonia bataticola. Out of the 24 rhizobial isolates tested only 2 isolates i.e., RPR17 and RPR24 showed inhibition activity against all the three pathogenic fungi. Bradyrhizobial isolate RPR16 showed maximum halo-zone to growth ratio i.e., 4.78 against Fusarium udum. Five rizobacterial isolates i.e., HPR49, HPR77, HPR80, HPR83 and HPR17 showed significant ACC utilization activity. Eight isolates i.e. HPR54, HPR58, HPR59, HPR65, HPR67, HPR71 and HPR83 produced IAA ranging from 12.3-26.31 μg/ml. Rhizobacterial isolates showed ALA production in the range of 0.1-23.45 μg/ml. Potassium solubilization studies showed that 28 isolates showed significant activity in the range of 4.1-6.0 and 42 rhizobacterial isolates showed KSI in the range of 2.1-4.0. The ability of siderophore production was found in only 41% of the rhizobacterial isolates. Significant cyanide production was observed in isolate HPR40 i.e., 76.54 μl/ml. Among the 83 rhizobaterial isolates, only six isolates i.e., HPR1, HPR36, HPR39, HPR15, HPR48 and HPR69 showed growth up to 8% NaCl salt concentration, whereas 26 isolates showed growth up to 4% salt with different colony size. Maximum increase in SDW (101.85%) was observed by co-inoculation of isolates RPR17+HPR78 followed by 97.68% increase in SDW by inoculation of isolate RPR17+HPR78 at 45 days of pigeon pea growth under chillum jar conditions. Coinoculation of isolates RPR17 + HPR78 and RPR17 + HPR17 resulted in 86.95 and 83.18% increase in RDW, respectively. Likewise, 108.59% increase in SDW was observed on inoculation of RPR17 + HPR78 isolates followed by 118.3 and 94.05% increase in SDW on inoculation of RPR17+HPR17 isolates, respectively at 60th day of observation. The inoculation effect of four antagonistic i.e., HPR17, HPR40, HPR49 and HPR78, and two bradyrhizobial isolates i.e., RPR19 and MRH46 was studied under pot house conditions on pigeon pea growth. At 45 DAS, the coinoculation treatment of RPR19 + HPR40 alongwith F. udum showed maximum nodulation i.e., 26 nodules/plant and 79.59 and 98% increase in SDW and RDW, respectively. At 60 DAS, the coinoculation treatment of RPR19 + HPR17 showed maximum nodulation i.e., 31 nodules/plant and 37.10 and 21.70% increase in SDW and RDW, respectively. At 75 DAS, coinoculation of RPR19 + HPR40 showed maximum nodulation i.e., 52 nodules /plant and showed significant increase in root and shoot dry weight of pigeon pea under pot house conditions. Rhizobacterial isolates HPR78 showed 100% disease control of F. udum at different stages of pigeon pea growth under pot house conditions. Coinoculation treatments i.e., RPR19 + HPR49 showed 100% disease control of Fusarium wilt at different stages of pigeon pea growth under pot house conditions. Rhizobacterial isolates HPR40 and HPR78 were identified as Pseudomonas aeruginosa strain SSR207 and Paenibacillus polymyxa strain SSR2_207. These plant growth promoting rhizobacterial isolates can be used for the enhancement of crop productivity under salinity stress in field conditions.ThesisItem Open Access Evaluation of thermotolerant bacteria for growth and yield of summer mungbean (Vigna radiata L.)(CCSHAU, 2019) Parashar, Atul; Leela WatiMungbean (Vigna radiata (L.) Wilczek), also known as green gram, is one of the important pulse crops of India. Mungbean is grown during spring, summer and kharif season in Northern India, while grown during Rabi season in Southern India. Summer mungbean being a crop of very short duration is grown after the harvesting of wheat and before sowing autmn crops.Various environmental stresses hamper the productivity of mungbean among which temperature plays key role.Plant growth promoting bacteria can play important role in increasing crop productivity and relieving environmental stress.In the present study, mungbean rhizobial and non-rhizobial bacterial isolates available in Plant Microbial Interaction Laboratary Deptt. of Microbiology, CCSHAU, Hisar were assessed for temperature tolerance and various plant growth promoting traits viz.IAA production,Ammonia excretion, P-solubilization, Siderophore production, ACC utilization and HCN production at different temperatures. Out of nine rhizobial isolates, only five isolates and out of five non rhizobial bacterial isolates four isolates were able to grow up to 450C.Under in vitro conditions rhizobial isolates MRH59 and MRH46 and non rhizobial isolates E13 and E17 were adjudged as promising plant growth promoting thermotolerant bacteria. All thermotolerant rhizobial isolates were tested for nodulation efficiency under sterilized conditions. Maximum nodule no.(16.67/plant) was, observed in isolate MRH 59.On the basis of various plant growth promoting traits and nodulation test isolates MRH59,MRH46 ,E13 and E17 were selected for plant growth promotion of mungbean (MH-421) under pot culture as well as field conditions during summer season. Nodulation efficiency and seed yield was better when thermotolerant rhizobial isolate MRH 59 was coinoculated with thermotolerant nonrhizobial bacterial isolate E13 in comparison to coinoculation of Rhizobium (MB703 reference culture) and phosphate solubiliser P-36 (reference culture).Under field conditions seed yield was 27.84% higher on coinoculation of rhizobial isolate MRH59 and non rhizobial isolate E13 suggesting that thermotolerant bacteria exerted beneficial effect on mungbean due to multiple plant growth promoting traits.ThesisItem Open Access Biocontrol of root-knot nematode (Meloidogyne javanica) in brinjal (Solanum melongena L.) using plant growth promoting rhizospheric bacteria(CCSHAU, 2018) Antil, Sonam; Rakesh KumarEgg plant (Solanum melongena L.) or brinjal is a nightshade species belonging to family Solanaceae and is the second most important vegetable crop after tomato and shows high susceptibility to infection by the root knot nematode Meloidogyne javanica. Management of the root-knot nematodes via biological methods such as PGPR is a novel technology emerging for sustainable agriculture. In this context, a total of 63 isolates were isolated from nematode affected rhizospheric soil of brinjal, tomato crops and earthworm culture media. All the 63 isolates were characterized for various plant growth promoting traits. Out of 63, fifteen isolates produced siderophores and maximum production was observed by isolate LBB-1and KMT-2. Chitinase activity was observed in 14 isolates and isolates KMT-2 and KMT-8 showed maximum chitinase activity. Sixteen isolates were found to be positive for HCN production and maximum production was seen in isolates NAE-1, KMB-3, KMT-2 and KMS-6. Fifty seven isolates were capable of excreting ammonia and maximum ammonia excretion was observed in KMT-2 (4.8μg/ml) and LBB-1 (4.7 μg/ml). All the bacterial isolates except one were producing IAA and high level of IAA production was observed in KMT-1 (45.9 μg/ml), KMB-2 (38.8 μg/ml), KMB-4 (27.7μg/ml) and KMT-5 (26.4μg/ml). A total of 39 isolates solubilized phosphate and highest P-SI was observed in isolate KMT-1(2.0) and LBB-4 (1.7). Twelve isolates- NAB-4, NAS-1, NAE-1, LBB-1, LBS-1, KMB-3, KMT-2, KMT-4, KMT-5, KMT-8, KMS-3 and KMS-6 resulted in minimum hatching in egg masses and higher mortality rate in J2 of M. javanica (upto 92%). Based on all the screening tests, a total of 12 were selected for pot house experiment on brinjal crop. Rhizospheric microbial count increased in all treatments from 0 DAP to 45 DAP. Under pot house conditions, T14 (RDF +KMT-4), T15 (RDF +KMT-5), T16 (RDF + KMT-8) and T17 (RDF +KMS-6) showed maximum plant growth, reduced galls and eggs in roots, and final nematode population in soil.