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M. Sc. Dissertations

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2004 - 2009202010 - 201634
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Subject: Microbiology × End date: 2016 × Type: Thesis × Thesis Type: M.Sc ×
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Now showing 1 - 9 of 54
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    ThesisItemOpen Access
    Indole acetic acid production by Pseudomonas species and its effect on plant growth of green gram and black gram
    (CCSHAU, 2004) Om Prakash; Sindhu, S.S.
    In the present studies, IAA producing Pseudomonas strains CPS59 and MPS90 were mutagenized with transposon Tn5 using E. coli strain S17-1. The frequency of mutants with low amount of IAA production varied from 35.14 to 46.75% from Pseudomonas strains CPS59 and MPS90, respectively and only 3.43 to 3.75% mutants produced higher levels of IAA in comparison to respective parent strains in LB medium supplemented with tryptophan. Inoculation of mutants derived from Pseudomonas strain CPS59 showed stunting effect on root development of green gram seedlings at both 5 and 10 days of observation whereas mutants CPS59-138, CPS59-162 and CPS59-231 showed stimulatory effect on root growth at 5 days in black gram. Majority of the mutants caused stimulation of shoot growth at both 5 and 10 days of observation in comparison to uninoculated treatment. MPS90-derived mutants i.e. MPS90-39, MPS90-157, MPS90-102 and MPS90-106 showed slight stimulation of root growth whereas most of the mutants showed stunting effect on shoot growth at 10 days in black gram. Coinoculation studies of Pseudomonas mutants with Bradyrhizobium sp. strain S24 resulted in increased nodule number, nodule fresh weight and shoot dry weight in green gram and black gram under chillum jar conditions. Shoot dry weight gains in green gram after coinoculation with CPS59-derived mutants varied from 107-149 per cent and from 110-137 per cent with MPS90-derived mutants in comparison to Bradyrhizobium-inoculated plants at 60 days of growth. Similarly, shoot dry weight gains in black gram varied from 102-178 per cent and 105-198 after inoculation with mutants derived from CPS59 and MPS90, respectively. The stimulation effect on shoot dry weight in comparison to uninoculated control treatment varied from 280-390 per cent in green gram and 179-357 per cent in black gram. Four mutants CPS59- 138, CPS59-321, MPS90-133 and MPS90-51 showed more enhancements in nodule formation by Bradyrhizobium strain S24 in green gram whereas mutants CPS59-162, CPS59-64, MPS90- 102 and MPS90-280 caused more stimulation for nodule formation in black gram at 60 days of plant growth. Significant enhancements in shoot dry weights of green gram were observed by coinoculation with Pseudomonas mutants CPS59-138, CPS59-231, CPS59-321, MPS90-280, MPS90-14 and MPS90-145. Similarly, coinoculation of Bradyrhizobium strain with Pseudomonas mutants CPS59-162, CPS59-321, CPS59-214, MPS90-102, MPS90-280 and MPS90-51 showed significant gains in shoot biomass of black gram at 60 days of plant growth. The better performance of IAA over producing mutants in relation to nodulation and plant biomass indicated that IAA production by Pseudomonas strains is a beneficial trait for selection of rhizobacterial strains having plant growth promotion ability.
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    ThesisItemOpen Access
    Development of quality assurance method for biofertilizers using LACz marker
    (CCSHAU, 2004) Manoj kumar; Gera, Rajesh
    Biofertilizers are living microorganisms used to supplement nutritional requirement of plants. Strains of Azotobacter, Rhizobium, Azospirillum, Pseudomonas, Bacillus and Acetobacter have been developed as biofertilizers for various crops and are suppled as carrier based inoculants. The responses of biofertilizers are inconsistent and generally related to the poor quality and poor competitive ability of the inoculants. The quality control methods include standard plate count and most probable number (MPN) by plant infection. Both these methods are not very specific and require at least 10-30 days. Thus, the quality control methods are not strictly followed because producers are not willing to hold their product for 30 days and thus substandard biofertilizers are being supplied to the farmers. Thus, there is an urgent need to develop simple method for quality assurance of biofertilizers, which could be completed with in 6-12 hours. lacZ marked strains of Azotobacter (A-2), Rhizobium (T-19CL and PP1021L), Pseudomonas (P-35) and Bacillus (BPL-2) showed relationship between -galactosidase activity with viable cell number at different stages of growth in liquid medium. The Z buffer with 0.1% Triton X-100 was found to be the best diluent for enumeration of viable cell population in charcoal based inoculants. There was relationship in -galactosidase activity and viable cell number of all these above strains in charcoal based inoculants, which remained stable up to three months. Time for development of yellow colour of ONPG filter paper discs was correlated with viable cell number, thus providing the qualitative method for enumeration of viable cell number in the bioinoculants.
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    ThesisItemOpen Access
    Evaluation of phosphate solubilizing bacterial strains/mutants with improved phosphate solubilization and their effect on mustard crop
    (CCSHAU, 2005) Manu Dev; Kundu, B.S.
    Sixty eight isolates of phosphate solubilizing bacteria (PSB) from rhizosphere of mustard grown in different region of Haryana showed a large variation in P-solubilization on solid as well as in liquid media. Most of isolates fell in <50% class of P-solubilization while in liquid media in 50-100μg/ml. No correlation was recorded between P-solubilization on solid media and liquid media. The selected isolates 15M, 22M and 25M showed P-solubilization 273.0 μg/ml, 301.0 g/ml, and 276.0 g/ml in liquid PVK, while in NBRIP 288.0 g/ml, 314.0 g/ml and 286.0 g/ml, and were used for subsequent studies. These isolates were identified as Pseudomonas sp. based on morphological and biochemical characteristic. The selected isolates were mutagenesied with NTG (50g/ml) for high P-solubilization. One hundered forty four mutants were selected based on killing rate between 70-80%. These mutants were checked for P- solubilization in solid and liquid PVK and NBRIP medium. The PSE of 15M mutants on solid medium varied from18.2-98.6% on PVK and from 14.1-96.2% on NBRIP media. While PSE of 22M mutants varied from 12.4-98.1% on PVK and from 18.4-100.2% on NBRIP media. The PSE mutant 25M varied from 12.1-77.1% on PVK and 10.2-85.4% on NBRIP solid media. The pH of 15M mutants varied from 3.6-6.0 in PVK in NBRIP it varied from 3.4-5.8, while pH of 22M mutant in PVK varied from 2.9-5.8 in PVK, in NBRIP it ranged from 2.1-5.6. The pH of 25M mutants varied from 2.5-5.9 in PVK and 2.3-5.8 in NBRIP. The 15M mutants showed P-solubilization between 177.1-324.1 g/ml in liquid PVK, while 176.4-336.4 g/ml in liquid NBRIP. 22M mutants showed P-solubilization between 182.2-362.2 g/ml in liquid PVK from 201.3-380.1g/ml in NBRIP. 25M showed P-solubilization between 167.4-351.9 g/ml in liquid PVK, while 175.8-364.5 g/ml in NBRIP. Strains (15M, 22M and 25M) and mutants (15M2, 15M6, 22M28, 22M29, 25M11, 25M30) were selected on basis of P-solubilization for pot culture experiment on mustard cv. RH-30. Total bacterial count in rhizosphere increased with URP at 30 and 45 DAS, while decrease was observed at 60 DAS. The number of these bacteria improved with seed inoculation and was more with URP. The number varied from 10-78, 27-91, and 13-37 at 30, 45, and 60 DAS respectively. The maximum number was observed with 15M6 with URP (50x105 cfu/g) followed by 22M28 with URP (78x105 cfu/g) at 30 DAS. At 45 DAS the maximum number was observed with 22M28 with URP (91 x 105 cfu/g) followed by 15MP with URP (85 x 105 cfu/g) and at 60 DASnumber was observed with 22M28 with URP (37 x 105 cfu/g) followed by 22M29 with URP (27 x 105 cfu/g). The PSB count in the rhizosphere varied from 1-24, 2-20 and 1-11 at 30, 45 and 60 DAS, respectively. The maximum number was observed with 22M28 (24 x 103 cfu/g) followed by 15M6 (19 x 103 cfu/g) at 30 DAS. At 45 DAS the maximum number was observed with 22M29 (20 x 103 cfu/g) followed by 22M29 with URP (19 x 103 cfu/g). The mustard biomass was more with SSP and with URP compared to the control at 30, 45 and 60 DAS. There was increase at 30 and 45 DAS but a 60 DAS with SSP marginal increase was observed. Almost all the mutants with URP showed increase in biomass except 25M30. The maximum biomass was observed with 22M28 (0.682 g/plant) followed by 22M29 (0.456 g/plant) at 30 DAS while at 45 DAS in case of 22M28 with URP (2.114 g/plant) followed by 22M29 (1.651 g/plant). At 60 DAS the maximum plant dry biomass was observed in 22M29 (3.161 g/plant) followed by 22M28 (2.927 g/plant). Phosphate uptake was increased upto 21 and 11% with application of SSP and URP at 30 DAS respectively but at 45 DAS increase was 12% with SSP and 4% with URP. But at 60 DAS there was 53% increase with SSP and 50% with URP. The maximum P-uptake was observed with 22M28 with URP (529%) followed by 22M29 is (429%). However, at 60 DAS the maximum P-uptake was by 22M29 (31%) followed by 22M28 with URP that was 308% it shows that mutants had significant effect on increase in plant dry biomass and P-uptake under pot house conditions.
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    ThesisItemOpen Access
    Isolation and identification of bacteria having pathogenic interactions with termites (Isoptera)
    (CCSHAU, 2007) Yuvraj singh; Sindhu, S.S.
    In the present study, the population of bacteria in termite nest soil varied from 1.2 x 106 to 90.0 x 106 colony forming units/g soil on nutrient agar medium. On soil extract agar medium, the bacterial population varied from 28.3 x 106 to 103.2 x 106 cfu/g soil and it varied between 2.6 x 106 to 256.0 x 106 cfu/g soil on King’s B medium. Based on colony morphology, gum production, colony characteristics and pigment production, bacterial isolates NNY 1-NNY 84 were selected from nutrient agar medium, isolates NSY 1-NSY 40 from soil extract agar medium and isolates NKY 1-NKY 96 were selected from King’s B medium. From the gut of termites, bacterial isolates GNY 1-GNY 20 were selected from nutrient agar medium plates. Isolates GSY 1-GSY 10 were obtained from soil extract agar medium and isolates GKY 1-GKY 20 were obtained from King’s B medium. A total of 270 bacterial isolates, i.e. 220 isolates from termite nest soil and 50 isolates from termite gut, were screened for the potential to produce different enzymes, i.e. lipase, protease and chitinase on specific media. In the preliminary screening, only 83 isolates were found to express one or more of the enzyme activities. Seven bacterial isolates without any enzyme activity were also selected. For lipolytic activity, the clearance zone to colony growth ratio varied from 1.24 to 4.20 in different bacterial isolates. Bacterial isolates NNY 73 and NSY 19 showed very little lipase activity and eight isolates did not show any lipase activity. Three isolates NNY 19, NNY 39 and NSY 20 showed 3.50, 4.20 and 4.00 enzyme production efficiency, respectively. Proteolytic enzyme production in different cultures varied from 1.24 to 2.29. The bacterial isolates NNY 24, NNY 50 and NNY 62 showed highest ratio of clearance zone to colony size i.e. 2.17, 2.14 and 2.29, respectively. Fourteen bacterial isolates did not show any proteolytic activity. Chitinolytic activity varied from 1.24 to 2.67 in different bacterial isolates. Isolates NNY 18, NNY 52, NNY 73 and GKY 10 showed very little chitinolytic activity. Four isolates, NNY 23, NNY 43, NNY 34 and NKY 91 showed 2.67, 2.50, 2.40 and 2.50 enzyme production efficiency, respectively. The bacterial isolates NNY 58, NKY 48 and NKY 69 expressed only chitinolytic activity. Isolate NKY 66 expressed only proteolytic activity whereas isolates NKY 17, NKY 62 and NKY 79 expressed only lipolytic activity. Twelve bacterial isolates were found to express all three enzyme activities. During studies of pathogenic interactions, only 47 bacterial isolates were found to kill the termites at 5 day of observation. The killing frequency of different bacterial isolates varied from 5.7 to 100 per cent at 5 day. Bacterial isolates NNY 23, NSY 19 and NKY 83 caused 100 per cent killing of the termites whereas 14 other isolates caused more than 82 per cent killing at 5 day. At 7 days of observation, 100 per cent killing of the termites was observed with four more bacterial isolates NNY 19, NNY 43, NSY 3 and NKY 91. Bacterial isolate NNY 23 possessed all the three enzyme activities and caused 100 per cent killing of the termites at 5 day of observation. Another bacterial isolate NKY 64 which did not show any of the enzyme activity but it caused 57.8 per cent killing of termites at 5 day. These results indicated that besides the production of three enzymes, some other metabolites (toxin or siderophore) could also be contributing to the killing of termites. Reisolation of bacteria from dead termites showed that seven bacterial isolates viz. NNY 52, NNY 62, NSY 3, NSY 10, NKY 25, NKY 27 and NKY 31 killed even the gut bacteria of the termites and only colonies of inoculated pathogenic bacteria appeared on nutrient agar, soil extract agar and KB medium plates. In the study of other termites inoculated with other 16 bacterial isolates, more than two types of colonies were observed on medium plates indicating that gut bacteria were not killed after ingestion/infection of antagonistic bacteria. Twenty-one bacterial isolates having more than 80 per cent killing of termites at 7 day of observation were identified by standard procedures up to genus level as described in Bergey’s Manual of Determinative Bacteriology. Gram-positive staining was observed only in bacterial isolates NNY 10, NNY 34, NNY 50, NNY 52, NSY 2, NSY 3, NSY 19, NKY 83, NKY 91 and GNY 17. Based on the comparative analysis of various morphological (colony morphology, pigment production and sporulation) and biochemical characteristics (such as indole production, methyl red test, Voges-Proskauer reaction, citrate utilization, oxidase, catalase and hydrolysis of starch and cellulose), the antagonistic bacteria were found to belong to the genera of Arthrobacter, Bacillus, Chromobacterium, Enterobacter, Micrococcus, Neisseria, Pseudomonas and Serratia.
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    ThesisItemOpen Access
    Utilization of pearl millet for ethanol production
    (CCSHAU, 2007) Poonia, Bijender; Leela Wati
    Ethanol is a known feed stock for countless chemicals and potable purposes. Recently the use of ethanol for fuel purpose is catching up faster world over. It is produced by fermentation of molasses by Saccharomyces cerevisiae in Indian distilleries. Molasses has become more expensive due to its decontrol and deteriorated in quality. Therefore, an alternate substrate that would be both cheaper and readily available in quantities, need to be sought and examined. Pearl millet grains were considered as potential substrate for ethanol production because of high starch content (56.9%). xv The optimum conc. of pearl millet flour slurry for liquefaction by a-amylase was found to be 30% which was liquefied by 600ml of enzyme at 85ºC in 10 min under shaking conditions (140 rpm). Saccharification of liquefied starch of pearl millet hybrids by spirizyme (750ml) at 60ºC under shaking conditions resulted in production of maximum 15.9% (w/v) reducing sugars in 2h. Fermentation of hydrolysate of various pearl millet cultivars at 30±2ºC by S. cerevisiae at pitching rate of 0.5% (w/v) generated 10.6-11.4% (v/v) ethanol with in 36 hrs. under stationary conditions. No significant effect of supplementation of hydrolysate with urea, peptone and yeast extract was observed for ethanol production suggesting that the pearl millet flour has sufficient N and P required for ethanolic fermentation.
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    ThesisItemOpen Access
    Isolation and characterization of yeast for glucoamylase production
    (CCSHAU, 2007) Nisha; Kundu, B.S.
    Glucoamylase is the major enzyme required for complete hydrolysis of starch to glucose, which can be used as carbon source in fermentation for ethanol production. It also has large application in food industries and distilleries. The demand for glucoamylase has increased many folds due to utilization of non-conventional substrates (starchy raw material) for industrial ethanol production. The screening of organism that produce large amount of enzyme has been a major area to improve the efficiency of starch processing. Nineteen starch assimilating yeasts were isolated from various sources (ripened fruits, spoiled vegetables, bakery samples, soil, unfermented wort and honey etc). Screening of glucoamylase producing yeast was done by starch plate iodine test and estimating glucoamylase activity. NY-19 and NY-3 isolated from unfermented wort and banana produced 9.2 and 9.6 IU/ml glucoamylase and were finally selected along with Saccharomycopsis fibuligera MTCC-3816 (reference strain). The reaction of 400 ml of cultural filtrate with 1% starch substrate at pH of 4.8 and 30oC temperature for one minute was found optimum for enzyme assay. At optima temperature (30oC), pH (5.0) glucoamylase production by NY-19, NY-3 and MTCC-3816 were 14.40, 13.00 and 15.97 IU/ml, respectively after 48 h of incubation under shake condition. Starch (3%), yeast extract (1%) and peptone (2%) were best carbon and nitrogen source respectively. However MTCC-3816 gave higher activity at 2% starch. The glucoamylase produced from NY-19 was partially purified to 10 fold with specific activity of 43.33 IU/mg of protein using (NH4)2SO4 fractionation. The partially purified enzyme has optimum temperature of 500C and pH 4.8 and was thermostable up to 400C for 8 h. 87% saccharification of rice flour slurry (30%) attained under optimum conditions of temperature (500C) and pH (4.5) in 2 h using Palkozyme HT Plus (liquefying enzyme) and glucoamylase preparation form NY-19. On the basis of morphological and biochemical characteristics isolate NY-19 was identified as Endomycopsis sp.
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    ThesisItemOpen Access
    Isolation and characterization of poly--hydroxybutyrate producing Bacillus sp. from soils
    (CCSHAU, 2008) Verma, Nisha; Goyal, Sneh
    Plastics and polymers are an integral part of our daily existence because of their properties of durability and resistance to degradation. Plastic wastes are considered to be severe environmental contaminants causing waste disposal problems. Recently, the problems concerning global environment and solid waste management have created much interest in the development of biodegradable plastics with desired physical and chemical properties. The PHB and other biodegradable polyesters are promising candidates for the development of environment friendly, totally biodegradable plastics. Polyhydroxy-alkanoates (PHAs) are intracellular carbon and energy reserves that are accumulated by many bacteria when surplus carbon is available, under certain nutrient limited conditions of growth. Different biodegradable plastics have been developed by microbial fermentations. However, limitations of these materials still exist due to high cost of production. For minimization of cost for the production of biodegradable plastics, the waste biological sludge generated at waste water treatment plants is one of the substrate used for the production of PHB. These compounds can be produced in large quantities from renewable resources by well known fermentation processes and have potential to replace petroleum derived thermoplastics. A total of 115 bacterial isolates differing in their morphology were isolated from various soils and were screened using Nile blue A and was verified by using Sudan black B dye. Nine PHB producing bacilli were selected depending upon their fluorescence on Nile blue A plates and PHB production under minimal medium broth was studied. High PHB producing Isolates ZTL-31, ZTL-32 and Sld-110 were selected for subsequent studies. Conditions for higher PHB production were optimized for temperature, time, aeration conditions and different carbon and nitrogen sources. Three different carbon sources milk whey, sewage sludge, molasses were used. Isolate No. Sld-110 produced highest amount of PHB (322.2 mg/g dry weight of cells) when 1 % of molasses was used as carbon source at 27C after 72 h under stationary conditions. Isolate Sld-110 produced up to 310.6 mg/g (w/w) PHB with ammonium phosphate as nitrogen source. However, it produced 356.4 mg/g (w/w) PHB without nitrogen source in minimal medium containing 1 % molasses at 27ºC after 72 h under stationary conditions.
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    ThesisItemOpen Access
    Development of microbial inoculant for the growth of Ashwagandha (Withania angustifolia)
    (CCSHAU, 2007) Dinesh Kumar; Yadav, K.S.
    A total of 104 isolates of rhizobacteria were obtained from rhizosphere and rhizoplane of Ashwagandha plants collected from three locations of Haryana. Of these 36 were from rhizosphere and 68 were from rhizoplane. All the isolates were screened for their growth promoting activities in terms of biomass production in sandy soil without addition of any organic or inorganic fertilizers. Inoculation with these isolates produced more plant biomass than control to a varying level. Only four isolates (HRP-7, RRP-8, and RRP-26 & YRP-11) produced plant biomass more than 10 g /plant. Highest plant biomass was produced by inoculation of isolate YRP-11 (16.49 g/plant) followed by RRP-26 (15.86 g/plant). Isolate HRP-7 produced least plant biomass. Isolates selected on the basis of higher plant biomass production showed useful traits like nitrogen fixation, and production of growth promoting substances like indole acetic acid. Inoculation with selected isolates increased plant biomass more in presence of farmyard manure than without farmyard manure. Mixture of these isolates produced biomass at par with mixed biofertilizer formulation containing Azotobacter and phosphorus solubilizing bacteria.
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    ThesisItemOpen Access
    Enrichment of vermicompost with agriculturally beneficial microorganisms and its impact on horticultural plants
    (CCSHAU, 2008) Khare, Neetu; Pathak, D.V.
    Vermicompost was enriched with Azotobacter chroococcum, Pseudomonas spp. P-36 , Pseudomonas maltophilia PM4 and AM fungi (Glomus mossae). The microbially enriched vermicompost was analysed for organic C, total N, total P, ammonical N and nitrate N and microbial population. Organic C was decreased in the microbially enriched vermicompost after 30 days due to microbial activity. Total N and total P content increased due to inoculation of Azotobacter chroococcum, Pseudomonas spp., and AM fungi (Glomus mossae). Microbially enriched vermicompost showed higher count of Azotobacter chroococcum, Pseudomonas spp., Pseudomonas maltophilia upto 30 days. Seedling of Aonla and Bael grown in microbially enriched vermicompost for 60 days showed increase in shoot and root length and plant dry weight. Seedling grown in AM fungi (Glomus mossae) enriched vermicompost had higher infection of AM fungi (Glomus mossae) as compared to control or vermicompost enriched with Azotobacter chroococcum, Pseudomonas spp., Pseudomonas maltophilia