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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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20215
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Subject: Microbiology × End date: 2021 × Start date: 2021 × Type: Thesis ×
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    ThesisItemOpen Access
    STRAIN IMPROVEMENT OF BACILLUS VELEZENSIS BY TRADITIONAL TECHNIQUES TO CONTROL PLANT PATHOGENS
    (Dr.RPCAU, Pusa, 2021) Rehman, Syed Vahida; Meena, Khem Raj
    Microbial 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.
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    ThesisItemOpen Access
    STUDY OF PLANT GROWTH PROMOTING ACTIVITIES OF ENDOPHYTIC BACTERIA ISOLATED FROM DIFFERENT VARIETIES OF SUGARCANE
    (Dr.RPCAU, Pusa, 2021) H S, SPANDANA; Jaiswal, Aman
    Endophytic 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.
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    ThesisItemOpen Access
    BIOCHEMICAL CHARACTERIZATION OF BACTERIA ISOLATED FROM DIFFERENT COMPOST SAMPLES AND GUT OF EARTHWORM USED IN VERMICOMPOSTING
    (DRPCAU, PUSA, 2021) KUMAR, SAMALA MANOJ; Jaiswal, Aman
    Composting 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.
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    ThesisItemOpen Access
    DEVELOPMENT OF PGPR BASED LIQUID INOCULANTS USING DIFFERENT WASTE PRODUCTS
    (DRPCAU, PUSA, 2021) SRAVANTHI, KUCHIPUDI; Jaiswal, Aman
    In 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.
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    ThesisItemOpen Access
    MICROBIAL BIOCONVERSION OF AGRI-WASTE INTO COMPOST: AN ECO-ENTERPRISING MODEL
    (DRPCAU, PUSA, 2021) S, AGILA; Meena, Khem Raj
    Bacterial 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.