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Dr. Rajendra Prasad Central Agricultural University, Pusa
In the imperial Gazetteer of India 1878, Pusa was recorded as a government estate of about 1350 acres in Darbhanba. It was acquired by East India Company for running a stud farm to supply better breed of horses mainly for the army. Frequent incidence of glanders disease (swelling of glands), mostly affecting the valuable imported bloodstock made the civil veterinary department to shift the entire stock out of Pusa. A British tobacco concern Beg Sutherland & co. got the estate on lease but it also left in 1897 abandoning the government estate of Pusa.
Lord Mayo, The Viceroy and Governor General, had been repeatedly trying to get through his proposal for setting up a directorate general of Agriculture that would take care of the soil and its productivity, formulate newer techniques of cultivation, improve the quality of seeds and livestock and also arrange for imparting agricultural education. The government of India had invited a British expert. Dr. J. A. Voelcker who had submitted as report on the development of Indian agriculture. As a follow-up action, three experts in different fields were appointed for the first time during 1885 to 1895 namely, agricultural chemist (Dr. J. W. Leafer), cryptogamic botanist (Dr. R. A. Butler) and entomologist (Dr. H. Maxwell Lefroy) with headquarters at Dehradun (U.P.) in the forest Research Institute complex.
Surprisingly, until now Pusa, which was destined to become the centre of agricultural revolution in the country, was lying as before an abandoned government estate. In 1898. Lord Curzon took over as the viceroy. A widely traveled person and an administrator, he salvaged out the earlier proposal and got London’s approval for the appointment of the inspector General of Agriculture to which the first incumbent Mr. J. Mollison (Dy. Director of Agriculture, Bombay) joined in 1901 with headquarters at Nagpur
The then government of Bengal had mooted in 1902 a proposal to the centre for setting up a model cattle farm for improving the dilapidated condition of the livestock at Pusa estate where plenty of land, water and feed would be available, and with Mr. Mollison’s support this was accepted in principle. Around Pusa, there were many British planters and also an indigo research centre Dalsing Sarai (near Pusa). Mr. Mollison’s visits to this mini British kingdom and his strong recommendations. In favour of Pusa as the most ideal place for the Bengal government project obviously caught the attention for the viceroy.
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ThesisItem Open Access STRAIN IMPROVEMENT OF BACILLUS VELEZENSIS BY TRADITIONAL TECHNIQUES TO CONTROL PLANT PATHOGENS(Dr.RPCAU, Pusa, 2021) Rehman, Syed Vahida; Meena, Khem RajMicrobial Biological Control Agents (MBCAs) are used to biologically control plant diseases in crops. The first step towards developing an efficient biological control is to identify effective antagonistic strains. Molecular approaches can be used to improve the ability of wild type strains to reduce disease severity and disease incidence of plants. Random mutagenesis induced by physical mutagens like UV, X-rays, and gamma radiation, as well as chemical mutagens like ethyl methane sulfonate have been used to modify organisms genetically. To improve the antagonistic nature of Bacillus velezensis a gram positive, endospore forming, biosurfactant producing bacterium, random mutagenesis has been used. A total 11 mutants were obtained when subjected to UV irradiation and ethyl methane sulfonate for different time intervals on the basis of random morphological characters. Out of all these mutants, 7 mutated isolates have shown biosurfactant production when screened through emulsification activity. The highest emulsification activity against hydrocarbons, benzene and toluene was shown in Luria Bertani broth media of 85.67±0.21 against benzene 85.88±0.20 against toluene. The crude yield of biosurfactant produced by mutants was also improved after mutagenesis. The highest yield of crude biosurfactant of 0.361±0.001 was shown by mutant M3 and followed by mutant M5 of crude biosurfactant yield was 0.332±0.002 Growth profile of all the mutants along with unmutated culture was studied. Some mutants (M3, M7, M9) had took nearly ~36 h to reach stationary phase as the cultures had taken much time in lag phase. The total protein content of all the strains were also analyzed. The highest protein content of 80.88μg was shown by mutant M3 followed by 77.74μg by mutant M5. All the mutants were screened for antagonistic activity against the selected plant pathogens Helminthosporium maydis and Fusarium oxysporum f.sp. lycopersici by dual culture plate technique. 7 mutants have shown nearly 60-80% inhibition against both the pathogens. Crude biosurfactant produced by the cultures were used to screen the pathogens with improved antagonistic activity. Six mutants M3, M4, M5, M6, M7, M9 were shown the improved antifungal activity against the selected pathogens. The highest antagonistic activity of 81.69 % and 83.33 % was shown by mutant M3 against Fusarium oxysporum f.sp. lycopersici and Helminthosporium maydis. These six mutants were selected further to check the biocontrol ability against the selected pathogens through pot experiment. Separate pot experiment was conducted to check the biocontrol ability against Fusarium oxysporum f.sp. lycopersici in tomato crop and Helminthosporium maydis in maize crop. The results obtained has showed the significant increase in growth of shoot length, shoot fresh and dry weight, root length, root fresh and dry weight of maize and tomato crop. It was observed that M3, M5, M7, M9 has shown the maximum enhanced growth parameters than unmutated strain in both maize and tomato crop. The disease severity index and disease incidence were calculated against southern corn leaf blight disease and fusarium wilt disease respectively. Results obtained were shown that M3, M5, M7, M9 has significantly reduced the disease severity in maize crop and mutants M3, M5, M9 has significantly reduced the disease incidence from 65.5 % in control to 29.7%, 32.4%, 33.1% respectively. The present study has been isolated the mutants with improved antagonistic activities could be used as potential biocontrol agents.ThesisItem Open Access STUDY OF PLANT GROWTH PROMOTING ACTIVITIES OF ENDOPHYTIC BACTERIA ISOLATED FROM DIFFERENT VARIETIES OF SUGARCANE(Dr.RPCAU, Pusa, 2021) H S, SPANDANA; Jaiswal, AmanEndophytic bacteria colonizing internal parts of the plants benefit the crop plants by enhancing their growth and development. These beneficial organisms are documented in almost all plants. Sugarcane also colonizes diverse endophytic bacteria and these bacteria benefit the sugarcane crop through plant growth promoting activities. Therefore, a study was conducted to isolate endophytic bacteria from different parts of two sugarcane varieties, BO-91 and Co-P 16437, and to examine them for plant growth promoting activities. Sugarcane samples, roots, stems and leaves were collected and further subdivided into upper, middle and lower portions. One gram of the sample from each sub portion was surface sterilized and subjected to an isolation process on nutrient agar and Luria Bertani agar plates. A total of 71 endophytic bacteria of different morphotypes were isolated from roots, stems and leaves of both varieties, out of which 45 isolates were obtained from BO-91 and 26 isolates from Co-P 16437. Isolates were subjected to morphological characterization on the basis of colony morphology and microscopic features. Most of the isolated colonies were of small size, circular shape with regular margins, smooth texture and yellow or orange color colonies on nutrient agar and Luria- Bertani agar plates. 41 isolates were identified as Gram-negative and 30 isolates as Gram-positive bacteria, out of which 17 were endospore producers based on Gram staining, endospore staining and microscopic observations and most of the isolated cells were of rod shape. Isolates were further tested for biochemical activities, in which most isolates were able to utilize citrate and showed positive response to the oxidase test, whereas few isolates showed positive response to the methyl red test and Voges-Proskauer test. Endophytic bacteria were subjected to plant growth promoting activities in vitro on the basis of nutrient solubilization (P, K and Zn) and production of useful compounds (siderophore and IAA). 16 isolates of BO-91 and 7 isolates of Co-P 16437 were able to solubilize phosphorus, in which the BS8 isolate of the stem part of BO-91 showed the highest value of P-SI (4.22) with a quantitative value of 42.66 ± 1.1 μg mg-1. 13 isolates of BO-91 and 8 isolates of Co-P 16437 were able to solubilize potassium, in which the CS6 isolate of stem part of Co-P 16437 showed the highest value of K-SI (3.16) with a quantitative value of 129.54 ± 0.8 μg ml-1. 12 isolates of BO-91 and 6 isolates of Co-P 16437 were able to solubilize zinc, in which the BR17 isolate of the stem part of BO-91 showed the highest value of Zn-SI (3.43) with a quantitative value of 119.68 ± 0.9 μg ml-1. 4 isolates of BO-91 and 2 isolates of Co-P 16437 were able to produce siderophores, in which the CR9 isolate of the root part of Co-P showed the highest value of SE (179%). 19 isolates of BO-91 and 10 isolates of Co-P 16437 were able to produce IAA, in which the CL2 isolate of the leaf part of Co-P 16437 showed the highest IAA production value of 29.70 ± 1.5 μg mg-1 protein. The BS16 isolate of stem part of BO-91 showed a positive response to phosphorus solubilization (P-SI of 3.13), potassium solubilization (K-SI of 2.19), zinc solubilization (Zn-SI of 2.30) and IAA production (20.83 μg mg-1 protein) activity. The CR9 isolate of the root part of Co-P 16437 showed positive indications of phosphorus solubilization (P-SI of 3.08), potassium solubilization (K-SI of 2.66), zinc solubilization (Zn-SI of 2.74), siderophore production (SE of 179%) and IAA production (28.15 μg mg-1 protein) activity. Thus, a diverse group of endophytic bacteria are present in different parts of the sugarcane crop and are involved in plant growth promoting activities. Therefore, the promising endophytic bacteria can be used as a biofertilizer which enhances sugarcane growth and development.ThesisItem Open Access BIOCHEMICAL CHARACTERIZATION OF BACTERIA ISOLATED FROM DIFFERENT COMPOST SAMPLES AND GUT OF EARTHWORM USED IN VERMICOMPOSTING(DRPCAU, PUSA, 2021) KUMAR, SAMALA MANOJ; Jaiswal, AmanComposting and vermicomposting are two examples of biological waste disposal systems that are generally regarded as environment friendly and long-term solutions for managing organic waste. Composting mechanism is regulated by complex community of microbes dominated by various bacteria. Earthworms are the driving force behind vermicomposting as the gut of an earthworm is a factory to manufacture the beneficial microbial density and their products to excrete thousand times more to enrich the surrounding soil. Proper understanding of microbiology and biochemistry should be deciphered. Therefore, in present study, bacteria were isolated from three different samples viz., mushroom compost, vermicompost and earthworm gut, which were collected from AICRP on Mushroom, Vermicomposting unit of Dr. Rajendra Prasad Central Agricultural University, Pusa on different media - Nutrient Agar, Trypticase soya agar, Ken knight agar, CMC agar and Kings B media. 20 isolates were obtained from compost, 40 isolates from vermicompost and 20 from earthworm gut. These isolates were checked for their staining ability viz., gram and endospore staining and found that 26 were gram positive, 54 gram negative and 11 were endospore formers. These isolates were further screened for cellulose and hemicellulose degradation activity and found that 10 isolates from mushroom compost, 19 from vermicompost and 8 isolates from earth worm gut were able to perform cellulose degradation activity whereas 8 from compost, 25 vermicompost isolates and 14 isolates from earthworm gut were able to degrade hemicellulose. These isolates were subjected to nutrient solubilization activities like phosphorous, potassium, zinc and siderophore production. 5 from compost, 17 from vermicompost and 10 isolates of earthworm gut respectively showed phosphorous solubilisation whereas 5 from compost, 16 from vermicompost and 11 isolates from earthworm gut showed potassium solubilization. In case of zinc solubilisation 10 from compost, 18 from vermicompost and 14 isolates from earthworm gut. 8 from compost, 21 from vermicompost and 12 from earthworm gut showed siderophore production. One isolate (MC-10) from mushroom compost and seven isolates from vermicompost showed all the six activities whereas none of the isolates from earthworm gut showed all activities together. However, some isolates showed a minimum of three activities and maximum four activities. In term of solubility indices, it was observed that earthworm gut isolates showed the highest Phosphorous, potassium and zinc solubility indices when compared to compost and vermicompost isolates. Those isolates which showed either of cellulose or hemicellulose activity and any three nutrient solubilization activities were further checked for biochemical reactions i.e., IMViC test. A total of 31 promising isolates were subjected to IMViC test in which 15 isolates showed positive for indole test. 16 showed MR test and 24 isolates showed VP test. 14 isolates were able to use citrate as a source of energy. Therefore, the promising bacterial isolates which showed cellulose and hemicellulose degradation activity and other nutrient solubilizing properties can be utilized in composting which will help in better and faster decomposition of waste and reduces the composting time period.ThesisItem Open Access DEVELOPMENT OF PGPR BASED LIQUID INOCULANTS USING DIFFERENT WASTE PRODUCTS(DRPCAU, PUSA, 2021) SRAVANTHI, KUCHIPUDI; Jaiswal, AmanIn the present study, 32 PGPR based liquid inoculants were prepared using different combinations of carrier (nutrient broth, water + Oil), emulsifier (Tween-80/Triton-X-100), thickeners (carboxy methyl cellulose, corn cob powder and sugarcane bagasse), pH maintainer (eggshell powder) and protective agents (polyvinylpyrolidine, glycerol and gum arabic) among those 24 were nutrient broth based and 8 were water based liquid inoculants. PGPR consortium was added into these formulations, which was prepared with six bacterial cultures viz., Bacillus sp. (BS5), Pseudomonas sp. (PS2, PS3), Azotobacter sp. (AZT1, AZT2) and Azospirillum sp. (AZS3) after finding that they were compatible with each other. Further the individual bacterial cultures and the developed PGPR consortium were screened for plant growth promoting characters viz., phosphorus solubilization, Zn solubilization, siderophore production and IAA production. It was found that plant growth promoting characteristics of PGPR consortium (PSI-3.9cm, ZnSI-3.54cm, IAA-163 μg mg1 protein and siderophore production efficiency- 257%) was higher than the individual cultures. The developed liquid inoculants were stored at three different temperatures such as 4°C, 28°C and 40°C. They were characterized for shelf life (cell count), suspensibility and pH at 1month interval up to 8months. After 8months of storage, the formulation stored at stored at 4°C showed highest cell count,stable pH and low suspensibility followed by formulations stored at 28°C and 40°C in both nutrient both based and water based formulations.When compared to the water based formulations, nutrient broth based formulations showed more fluctuations in pH and cell count.But in terms of survivability nutrient broth based formulations showed highest cell count as compared to water based formulations after 8months of storage period.Among the nutrient broth based formulations, those formulation where 75% nutrient brothwas used, showed highest shelf life and pH followed by 100% (NB) and 50% (NB). Shelf life was highest in the formulations prepared with all types of thickeners (CMC + CP + SB) followed by formulations with two types of thickeners and single thickener at all storage temperatures. Formulation F16 (NB-75% + T-80/T-100 + GA+ PVP + GLY+ EP + CMC+ SB + CP) showed highest cell count i.e., 63 x108 cells/ml, 59 x108 cells/ml and 56 x108 cells/mlat 4°C, 28°C and 40°C respectively. Whereas in case of water-based formulations, formulation F32 (Water + Oil + T-80/T-100 + GA + PVP + GLY + EP + CMC + SB + CP) showed highest cell population. Least cell population was observed in those formulations (F1, F9, F17 and F25) which were prepared without any thickeners. The formulations have been checked and found to possess a shelf life of 8months and this new PGPR based liquid formulations using waste products can play significant role in sustainable utilization waste management and help in restoring the soil health and productivity on long term basis with its availability and applicability.ThesisItem Open Access MICROBIAL BIOCONVERSION OF AGRI-WASTE INTO COMPOST: AN ECO-ENTERPRISING MODEL(DRPCAU, PUSA, 2021) S, AGILA; Meena, Khem RajBacterial isolates were collected from Department of Microbiology, Dr. Rajendra Prasad Central Agricultural University, Pusa. Total 28 isolates were collected and then subculturing were done in different media and broth. Based on 16S rDNA sequence analysis VC2, VC4, VC6, VC7, VC12, C10 and CC6 were identified as Bacillus sonorensis, Bacillus subtilis, Bacillus inaquosorum, Bacillus cabrialesii, Bacillus subtilis, Bacillus paramycoides and Bacillus stercoris. Out of 28 isolates, 27 isolates showed cellulose hydrolysing ability indicated by the formation of clearance zone in the range of 12.62 mm to 32.05 mm and the highest activity was shown by VC2 (32.05 mm). In the case of hemicellulose, 18 isolates showed positive results with hydrolysing zone formation in the range of 12.4 mm to 33.5 mm and the highest activity was shown by M18 (33.51 mm). As the enzyme activity concerned, cellulase production ability was reported by 16 isolates. Among them isolate VC2 had highest enzyme activity of 0.5428 U/ml with specific activity of 0.0037 U/mg and the lowest activity was indicated by C10 was 0.0668 U/ml with specific activity 0.0009 U/mg. Xylanase production ability was reported by 10 isolates, isolate VC7 showed highest enzyme activity 8.2479 U/ml with specific activity 0.0341 U/mg and the lowest activity was reported by C8 was 0.0368 U/ml. Bacterial strains that were observed with high enzymatic activity further tested for antagonistic activity. All the isolates were compatible with each other without showing any antagonistic behaviour. Compatible cultures were grown separately in enrichment media then they were mixed to produce bacterial consortia which was used for preparation of formulations. Formulations of different concentrations were prepared using various substrates like wheat straw (2g), glucose (6g), lime (3%), Urea (2%), microbial consortia and distilled water of varying quantities but final volume was made to 200 ml. 18 formulations were prepared with 3 replications. Further analysis of wheat straw for degradation was done using different formulations. There was observed that the pH of the formulations varies related to various concentrations of different substrates. Highest pH was observed in formulation containing lime and glucose it is because lime increases pH of the solution. A15 formulation showed showed highest pH (11.13) followed by A3 (10.67) then the lowest pH was shown by A16 (2.54). It was observed that all the formulations at the initial showed decreased pH until 14 days then increased upto 42 days then decreased to attain pH constant (56 days). pH at the initial stage was high (>7.5) found to have less amount of nitrogen. At the next stage (i.e., after 20- 24 days) there occurs sharp increase in nitrogen content due to reduced pH (<6.5) causes increased accumulation of N and at the end of composting, pH again starts increasing slightly to maintain optimum level (i.e.,6.5 - 7) but N content slightly decreased or increased based on nutrient availability and decomposition rate. High N content was reported in formulation containing microbial consortia with pH in optimum level (i.e., 6.5-8). Formulations maintaining pH range 6.5 to 8 were observed to produce optimum N content and the formulations containing pH in the too acidic or alkaline range leads to contain very less or no N content.ThesisItem Open Access Study of microbes in button mushroom compost production(DRPCAU, Pusa, 2020) Kumar, Chandra Mohan; Meena, Khem RajMushrooms are rich source of nutrients and have exceptional medicinal properties. The most important commercially grown genus of edible mushroom is Agaricus bisporus (button mushroom) and A. bisporus (Lange) Imbach, which accounts 86% of the overall mushroom yield in India. The cultivation of mushroom requires nutritionally rich substrate i.e., compost but microbiology and biochemistry of composting is not understood properly yet. The microbial consortium can be help as a booster for bio-composting agricultural residues and fasten the rate of composting for mushroom production. Therefore, in the recent studies, the compost degrading bacteria were isolated from the button mushroom compost (degraded compost and spent mushroom substrate) which was collected from AICRP on mushroom of Dr. Rajendra Prasad Central Agricultural University, Pusa on different media viz. Nutrient Agar media, T3 Media, Kings‘s B agar media, Trypticase soya agar media and Kenknight agar media. A total of 40 isolates were obtained based on different colony characteristics and which were screened for cellulose and hemicellulose degradation activities. Out of the 40 isolates, 18 isolates showed cellulose degradation ability with the zone of clearance was in the range of 12.6 mm to 35.3 mm and isolate C-7 showed highest zone of clearance (35.3 mm). Among the cellulose degrader, 10 isolates were positive for hemicellulose degradation with the zone of clearance in the range of 12.4 mm to 33.5 mm and isolate M-18 showed highest zone of clearance (33.5 mm). Those isolates which showed both cellulose and hemicellulose degradation abilities were further characterized morphologically and biochemically by gram staining, endospore staining and IMViC test. It was observed that out of 10 cellulose and hemicellulose degrading isolates, 4 isolates viz. M-3, M-18, C-4 and C-6 were gram negative and 6 isolates viz. M-1, M-10, M-11, M-19, C-10 and C-12 were gram positive and among gram positive isolates, only two isolates viz. C-12 and M-1 showed endospore formation. Those isolates which showed positive result for cellulose and hemicellulose degradation were further assayed for IMViC test. It was observed out of 10 isolates, isolates M1, M6, M13 and C11were positive for Indole test whereas in Methyl red test, all the 10 isolates showed positive result. For Voges-Proskauers (VP) test, it was found that except 2 isolates viz. M6 and C19,all the isolates showed positive result whereas all the 10 isolates were negative for citrate utilization test. These ten isolates were also tested for antagonistic behavior for consortia formation and found that none of the isolates showed any antagonistic ability against each other. Therefore, these isolates were used for the preparation of microbial consortia by growing them in enrichment culture media which were further inoculated in developed formulations. Different formulations were prepared using different combinations of substrates i.e., lime (3%), urea (2%), glucose (10g) and distilled water at different concentration and then inoculated with microbial consortium and 3g sterilized wheat straw and stored at 280 C. These formulations were further characterized for pH, microbial growth (optical density) and cell count at different time interval i.e., 0 day, 14days, 28 days, 42 days and 56 days. A total of 18 formulations were developed with three replicates. It was reported that pH varied according to the substrates added to the formulations. The addition of lime increased the pH when compared to the other formulations. In almost all the formulations, the pH values initially (14 days) decreased then increased and became almost static at the end of incubation i.e., after 56 days. It was found that in all the formulations, where glucose was added, there was lowest pH recorded as compared to the other formulations. Formulation F18 showed highest pH (11.78) followed by F17 (11.32) when consortia was inoculated and least (2.65) in case of formulation F1. When the optical density (at 660 nm) of formulations were recorded at various time intervals, it was observed that, in all the formulations there was increase in optical density gradually and the optical density of the formulations did not show a specific pattern and varied independently. At 0 day, the optical density was highest i.e., 2.57 in formulation F18 followed by formulation F7 (2.26) and lowest was recorded in formulation F3 (0.68). Whereas at 56 days of incubation, formulation F16 showed the highest optical density (2.97) and the lowest was observed in formulation F3 (0.80). The cell count of formulations varied according to the change in the pH i.e., the cell count was higher at neutral pH as compared to the acidic and alkaline pH of the formulations. As compared to the initial cell population (6.3 X 106) added to each formulation, the cell population in most of the formulation declined initially up to 14 days and then increased and became static at 56 days of incubation. Formulation F5 showed the highest cell count (6.0 X 106) followed by formulation F7 and F8 (5.8 X 106) formulation F1 showed the lowest cell count (5.1 X 106) after 56 days of incubation. Therefore the developed microbial consortium based formulations can be used to hasten the rate of composting for button mushroom production.ThesisItem Open Access Evaluation of Tal Rhizobium strain for its efficacy under different soil system in Mono and consortium mode(DRPCAU, Pusa, 2020) Pathak, Devashish; Jha, M. N.Chemical fertilizer has to be supplemented with bio-fertilizer to prevent the deterioration of the soil quality, surface water, ground water pollution, reduction of biodiversity, imbalance microbial load and disturbance in the ecosystem functioning. Thus there is an immediate need to amalgate chemical fertilizer with bio-fertilizer to sustain the agricultural productivity. A major demand for bio-fertilizer industry is to supply of such microbial strain either in mono and consortia mode having wider agro-ecological application. Accordingly, this study was carried out with isolation of Rhizobium and rhizobacteria from lentil plant root and rhizosphere of Mokama Tal land (pulse basket of Bihar), having negligible chemical fertilizer application to formulate a microbial consortia for various plant promoting and nutrient solubilisation characteristics. Among 28 isolates, 12, 15, 26 and 9 isolates were tested positive for acetylene reduction assay, ammonia production, IAA production and HCN production respectively. Also 20, 21, 17 and 9 isolates were positive for phosphorus solubilisation, siderophore production, Zn solubilisation and potassium solubilisation respectively. The best isolates were screened for carbon source utilization and found DPR2, DPB4, DPP4 and DPA1 had diverse nature for C-source utilization, thus were selected for consortia preparation. Morphological and molecular identification of selected isolates DPR2, DPB4, DPP4 and DPA1 were done, and identified as Rhizobium leguminosarum bv. viciae, Bacillus teliquencis, Pseudomonas fluorescens and Azotobacter salinestris respectively. A pot experiment was carried out to evaluate the prepared consortia with respect to the chemical fertilizer in lentil plant in different soil conditions like Tal soil of Mokama, Diara soil of Begusarai, Chaur soil of Darbhanga and calcareous soil of Pusa. In plant growth and yield attributing characteristics, result of consortia for root length, 100 seed weight and yield per pot were higher in Tal soil among three other soil but for no. of branches, days of germination, no. of pods/plant and no. of seeds per pod were found insignificantly differ for Tal soil, Chaur soil and Diara soil followed by calcareous soil. In shoot length and no. of nodules/plant result of consortia were similar for Tal and Chaur soil followed by Diara soil and least for calcareous soil. In nutrient acquisition, result of consortia for nitrogen and Fe content in seed were highest in Tal soil followed by Chaur soil. In phosphorus and Zn content in seed, result of consortia was higher in Tal soil followed by Diara soil. Whereas for potassium content in seed, result of consortia were similar for Tal and Chaur soil followed by Diara soil. Among all 13 treatments, best result was found in T13 consortia (Rhizobium + Bacillus + Pseudomonas + Azotobacter + 50% RDF) compared with the 100%RDF and mono mode application of Rhizobium + 50% RDF, across the soil type. Thus, the consortia of Rhizobium + Bacillus + Pseudomonas + Azotobacter can be used as a universal strain under different soil system of Bihar for lentil crop production.ThesisItem Open Access Development of effective microbial consortia for secondary treatment of wastewater(DRPCAU, Pusa, 2020) Behera, Dhrupad Kumar; Jaiswal, AmanNaturally occurring bacteria present in the wastewaters have a massive potential to solve the problem of wastewater bio-remediation. These bacteria with biodegrading capabilities can be exploited to develop a consortium and can be used for efficient biological treatment of the wastewaters. In the present study, 20 bacterial strains were isolated from the dairy industry, sugar industry and hostel sewage water of Pusa campus from the state of Bihar and were screened for starch, protein and fat degradation and reduction Biological Oxygen Demand and Chemical Oxygen Demand. Out of the total 20 isolates, 18 isolates exhibited starch degradation, 10 exhibited protein degradation and 11 reported positive for the fat degradation. Among all these isolates, only 7 isolates (DS-13, DS-27, DS-57, DS-58, DS-67, DS-68, and DS-72) were found positive for all three degradation activities which were further screened for BOD and COD reduction. Out of the 7 isolates, only 3 isolates (DS-58, DS-67 and DS-72) showed higher reduction in both BOD and COD parameters when inoculated into autoclaved effluents i.e. dairy, sugar industry and hostel sewage as compared to the blank i.e., un-autoclaved effluents without inoculation of bacterial isolates which were recorded BOD 835 mg/L, 1035mg/L and 620mg/L and COD 1680 mg/L, 2280 mg/L and 1470 mg/L for the dairy, sugar industry and hostel sewage effluent respectively. Therefore, these 3 isolates were combined in different combinations and 4 different consortia were prepared and their biodegradation and BOD and COD reduction abilities were analysed. The consortium C4 (DS-58, DS-67 and DS-72) showed highest results in starch (14 mm), protein (24 mm) and fat (18 mm) degradation and BOD (115 to 353 mg/L) and COD (407 to 641 mg/L) reduction which was used further in the developed formulation. A total 10 liquid formulations were developed using several combinations of emulsifier (Tween 20), thickener (Acacia gum and CMC) and protective agents (PVP and glycerol) and stored at three different temperatures (40C, 280C and 400C) and characterized for physiological stability viz. pH and suspensibility and cell count at an interval of 30 days up to 180 days. In the case of pH, it was found that most of the formulations showed a slow fall in pH with increasing time of incubation. At 4ºC, the pH varied between 4.72 and 7.0 among different formulations during 6 months of incubation and formulations F7 and F8 were found most stable in terms of pH where the pH changes were of very less magnitude over time. At 280 C, the pH range was 4.74 to 7.0 and formulations F6 and F5 were found to be most stable whereas, at 400 C, the pH range was 4.71 to 7.0 and formulations F7 and F 8 were found to be most stable. In the case of Suspensibility, the width of suspension was inconstant and fluctuated from 0 to 1.2 cm between different formulations across different temperatures. Initially, it increased with increase in incubation period till 4 months and later decreased with further increase in the incubation period and became stable irrespective of the type of formulations at all the three stored temperatures. The reduction observed was comparatively more at a higher temperature of 400C than at a lower temperature of 40C. In case of the cell population, as compared to initial cell population of 6.8 x 107 cells/ ml that was added to each formulation, all formulations maintained a cell population of 2.6 x 107 to 5 x 107 cells/ ml after 6 months of incubation across all the stored temperature. It was also observed that the decline was also observed which was less during the early incubation period and decreased on further storage. After 6 months of incubation, F7 maintained maximum cell population of 5 x 107 cells/ml followed by F5 and F8 at 40C whereas, at 280C, formulation F6 and F8 maintained maximum cell population of 4.8 x 107 cells/ml followed by F4 which was 3.7 x 107 cells/ml. At 400C, formulation F8 and F7 maintained maximum cell population of 4.0 x 107 cells/ml closely followed by formulation F6 (3.7 x 107 cells/ml) after 6 months of incubation. It was found that formulation F10 i.e. formulation in which pH was not adjusted and no protective agents were added and only mixed cultures were inoculated showed minimum cell population at the end of six months at all the stored temperatures of 40C, 280C and 400C respectively. It was also observed that those formulations where the mixture of protective agents was used showed higher cell count as compared to those formulations where protective agents used as alone. The results suggested that this liquid formulation of consortium be able to play a significant part in the biological treatment of wastewater and help in reducing water pollution.ThesisItem Open Access Prospecting of multitasking strain with potential of biofertilization, biostimulation and biocontrol of Turcicum leaf blight of maize(DRPCAU, Pusa, 2020) Akhila, Pole; Jaiswal, AmanRhizobacteria have vital role in agriculture as they enhance the plant growth, biomass, yield, nutrient content and suppress the phytopathogens. These multitasking abilities of rhizobacteria were exploited to see their role in maize crop. A total 48 rhizobacterial isolates were isolated from soil samples collected from different regions of Bihar and were screened for various biofertilizing, biostimulating and biocontrolactivities viz., nitrogen fixation, phosphorus solubilization, potassium solubilisation, Zn solubilization, Siderophore production, HCN production, Ammonia production, IAA production, chitinase production and antagonistic ability against Turcicum leaf blight i.e., Exserohilum turcicum. Out of 48 rhizobacterial isolates,10 isolates showed nitrogen fixation, 37 were positive for P-solubulization, 22 were positive for K-solubulization, 33 were positive for Zn-solubulization, 29 showed siderophore production, 27 showed IAA production, 11 showed HCN production, 17 showed ammonia production, 19 showed chitinase production and 9 isolates showed antagonistic ability against Exserohilum turcicum. Among all these screened isolates, only two isolates i.e., AK-9 and AK-21were positive for all the activities. So, these two isolates were selected as multitasking strains and further used in pot experiments against maize crop. Two pot experiments were conducted. First pot experiment was conducted in maize variety-Shaktiman-5 to evaluate the biofertilization and biostimulation ability of these multitasking strains along with different levels of chemical fertilizers (N, P, K, Fe, Zn) on plant growth parameters (shoot length, root length, shoot fresh and dry weights, root fresh and dry weights), yield parameters (Number of grains per cob, 100 seed weight and yield per pot) and nutrient contents (N, P, K, Fe and Zn) of grain and plant. All these parameters were enhanced when multitasking strains were used in combination with different dosages of chemical fertilizer as compared to control and 100% RDF. Maximum enhancement was recorded in treatments containing 100%RDF+Fe+Zn+AK-9 followed by treatment containing 100%RDF+Fe+Zn+AK-21. Second pot experiment was conducted to evaluate the biocontrol ability of these multitasking strains along with different levels of fungicide in maize variety-Shaktiman-5 along with one susceptible variety CML-186 and plant growth and yield parameters, nutrient content and disease severity index were analysed. It was observed that there was enhancement in the crop growth, yield and nutrient contents and decrease in disease severity index in both the maize varieties when multitasking strains along with fungicide were applied as compared to control and 100 % fungicide.Disease severity index of the treatment containing Pathogen inoculation+50% Fungicide+AK-9/ AK-21was on par with the treatment containing Pathogen inoculation+100% Fungicide.Therefore, these two isolates AK-9 and AK-21, having multitasking ability, can play a significant role in biofertilization, biostimulation and biological control of maize crop against Turcicum leaf blight disease.
