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ThesisItem Open Access AEROBIC AND ANAEROBIC DIGESTION OF AGRICULTURAL WASTE FOLLOWED BY VERMICOMPOSTING AND ENRICHMENT(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY. HYDERABAD, 2016) PRASANNA KUMAR, B; Triveni, SIn the present study the experiment was conducted during 2015-16 at Department of Agricultural microbiology and Bioenergy, College of Agriculture, Rajendranagar, PJTSAU, Hyderabad. To expand the range of natural bio-resources the rapidly evolving tools of biotechnology can lower the conversion costs and also enhance target yield of the product of interest. Cellulolytic microorganisms have been isolated from different soil samples and also from waste samples characterized and screened for their efficiency in cellulose degrading activity. The best isolates were further tested for their enzyme activity. These efficient isolates were used for the pretreatment of agricultural waste (Maize straw) and horticultural waste (Banana fruit waste) prior to aerobic and anaerobic digestion determination. Cowdung along with agricultural waste and horticultural waste were used for composting, vermicomposting, biogas production and bioethanol production in lab scale. By the end of the process of aerobic composting and anaerobic digestion of agricultural waste the reduction percentage of Total Solids (TS) (14.99 %), pH (7.58), Chemical Oxygen Demand (COD) (190.82 g l-1), Cellulose (16.20 %), bacterial population count (6.4 x 106 CFU g-1) and actinomycetes (6.1 x 104 CFU g-1) was significantly more in T1 (Composting without pretreatment). Reduction of Organic carbon (31.32 %) was significantly more in T2 (Composting with pretreatment). Reduction in Total Volatile Solids (TVS) (76.48 %), Nitrogen (2.00 %) and Potassium (1.50 %) was significantly more in T3 (Vermicomposting without pretreatment). Reduction of Volatile Fatty Acids (VFA) (0.38 g l-1) was significantly more in T4 (Vermicomposting with pretreatment). Change in pH (7.58) was significantly more in T5 (Biogas production without pretreatment). Reduction in Phosphorus (1.65 %), Biological Oxygen Demand (BOD) (90.65 mg l-1) and fungal population count (3.8 x 104 CFU g-1) was significantly more in T6 (Biogas production with pretreatment). By the end of the process of aerobic composting and anaerobic digestion of agricultural waste the deterioration of Nitrogen (1.05 %), Potassium (0.68 %), Biological Oxygen Demand (BOD) (45.80 mg l-1) and fungal population count (2.1 x 104 CFU g-1) was significantly less in T1 (Composting without pretreatment). Reduction in Phsphorus (0.38 %) was significantly less in T2 (Composting with pretreatment). Reduction in Organnic carbon (22.52 %) and pH (7.21) and bacterial population count (3.5 x 106 CFU g-1) was significantly less in T4 (Vermicomposting with pretreatment). Reduction in Total solids (TS) (7.05 %), Total volatile solids (TVS) (45.29 %) and Chemical Oxygen Demand (COD) (59.46 g l-1) was significantly less in T5 (Biogas production without pretreatment). Reduction in Volatile fatty acids (VFA) (2.26 g l-1) and cellulose (12.85 per cent) and actinomycetes (4 x 104 CFU g-1) was significantly less in T6 (Biogas production with pretreatment). Similarly in horticultural waste by the end of the process of aerobic composting and anaerobic digestion the change in pH (7.90) and actinomycetes (7.2 x 104 CFU g-1) was significantly more in T1 (Composting without pretreatment). Reduction in Total Solids (TS) (16.15 %) and Chemical Oxygen Demand (COD) (96.25 g l-1) was significantly more in T2 (Composting with pretreatment). Reduction in Total Volatile Solids (TVS) (78.72 %), Biological Oxygen Demand (BOD) (85.15 mg l-1) and cellulose (17.36 per cent) was significantly more in T3 (Vermicomposting without pretreatment). Reduction in Phosphorus (1.58 %) and Potassium (1.56 %) was significantly more in T4 (Vermicomposting with pretreatment). Reduction in Organic carbon (45.7 %) and fungi (11 x 104 CFU g-1) was significantly more in T5 (biogas production without pretreatment). Reduction in Volatile Fatty Acids (VFA) (1.25 g l-1) Nitrogen (2.30 %) and bacteria (9.8 x 106 CFU g-1) was significantly more in T6 (Biogas production with pretreatment). By the end of the process of aerobic composting and anaerobic digestion of horticultural waste the deterioration of fungi (7 x 104 CFU g-1) was significantly less in T1 (Composting without pretreatment). The reduction in Nitrogen (1.30 %), Phosphorus (0.31 %), Potassium (0.60 %), Organic carbon (23.2 %) and bacteria (5.7 x 106 CFU g-1) was significantly less in T2 (Composting with pretreatment). Reduction in Volatile Fatty Acids (0.60 g l-1) was significantly less in T3 (Vermicomposting without pretreatment). Change in pH (7.33) was significantly less in T4 (Vermicomposting with pretreatment). Reduction in Total solids (TS) (6.99 %), Total volatile solids (TVS) (53.41 %), Biological Oxygen Demand (BOD) (49.45 mg l-1) and Chemical Oxygen Demand (COD) (59.39 g l-1) was significantly less in T5 (Biogas production without pretreatment). Reduction in cellulose (14.03 %) and actinomycetes (3.7 x 104 CFU g-1) was significantly less in T6 (Biogas production with pretreatment). At the end of the 60th day the methane gas production was significantly more in AW-T2 (Biogas production with pretreatment) 3531.10 ml, compared to AW-T1 (Biogas production without pretreatment) 3381.00 ml, HW-T2 (Biogas production with pretreatment) 2620.70 ml and less in HW-T1 (Biogas production without pretreatment) 2381.40 ml. The content of bioethanol was less in HW-T1 (Bioethanol production without pretreatment) 0.050 g L-1 compared to less in HW-T2 (Bioethanol production with pretreatment) 0.047 g l-1, AW-T2 (Bioethanol production with pretreatment) 0.036 g l-1 and less in AW-T1 (Bioethanol production without pretreatment) 0.034 g l-1. In this present study results revealed agricultural and horticultural waste pretreatment with efficient microbes played good role during aerobic composting, vermicomosting, and anaerobic bioethanol and biogas production capabilities. Hence, composting, vermicomposting, biogas production has proved to be effective and efficient in conversion of wastes into value added products. Hence it can be concluded that horticultural waste was comparatively better in N, P, K and Organic carbon % composition while agricultural waste was good for compost and vermicompost making and also useful for the production of biogas.ThesisItem Open Access BIODIVERSITY OF ALCOHOL PRODUCING YEAST- Saccharomyces cerevisiae(ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, 2011) PISE SATISH AMBADAS; SUMATHI, SEthanol is quantitatively and economically the world’s premier biotechnological commodity. Ethanol can be produced from any fermentable carbohydrates by yeasts. A long term goal of the brewing industry is to identify yeast strains with increased tolerance to the stresses experienced during the brewing process. A combination of high temperature, ethanol and sugar tolerance is clearly desirable characteristics in fermentation processes. Until 1990s, classical tests based on morphology, biochemical and sexual reproduction characteristics were used for wine yeast identification. Such methods only allowed discrimination between species and they were laborious, time consuming and imprecise. In the last decade, the application of several molecular techniques has generated a large number of studies concerning ecology and biodiversity of indigenous Saccharomyces cerevisiae strains. The present investigation is carried out with the objective of isolating ethanologenic yeasts from different sources and location followed by screening for diversity on basis of alcohol production and functional characteristics using cultural and molecular methods. Twenty samples were obtained from different sources and pure isolates were identified as yeast cultures based on their morphological characters, cultural characters, carbohydrate assimilation profile and alcohol producing ability. All isolates had alcohol producing ability but maximum alcohol production was observed with standard yeast culture Saccharomyces cerevisiae MTCC-172 (67.77 g l-1) followed by isolate APY-4 (67.53 g l-1) and AMY-2 (67.50 g l-1) while isolate AMY-7 produced the least (30 g l-1). Twenty yeast isolates and standard yeast culture Saccharomyces cerevisiae MTCC- 172 were screened for ethanol tolerance, thermo tolerance, sugar tolerance and osmotolerance. It was found that isolates exhibited high diversity for tolerance and were grouped according to it. Based on these results it was observed that isolates APY-4, AMY- 2, AMY-9 and standard culture Saccharomyces cerevisiae - MTCC-172 were best among all isolates. These four pure cultures were tested for their alcohol producing ability at 40°C by inoculating them in fermentation medium containing 20% sugar. Four yeast cultures fermented sugar and produced alcohol at 40°C and 20% sugar concentration. Isolate AMY- 2 showed superior capacity for alcohol production and sugar conversion efficiency (SCE) while the standard culture Saccharomyces cerevisiae MTCC-172 exhibited better alcohol production and sugar conversion efficiency (SCE) at 40°C and 20% sugar concentration. Genomic DNA of 21 pure cultures of Saccharomyces cerevisiae were analyzed by RAPD technique using fifteen different primers and a dendrogram was constructed. Only isolates AMY-7 and APY-2 formed separate single element groups in dendrogram. The dendrogram analysis was compared with analysis formed from assimilation profiles, cultural characteristics, morphological characteristics and functional characteristics (sugar conversion efficiency, thermotolerance, osmotolerance, ethanol tolerance and sugar tolerance) which revealed great biodiversity among the twenty one pure Saccharomyces cerevisiae cultures. Based on the present study it is established that the important functional diversity found for ethanol tolerance, thermotolerance, sugar tolerance and osmotolerance indicate that the use of wider range of yeast isolates could be outlined and further developed for valorization. These results can be applied for optimization of alcohol production, brewery and baking industries as well in tropical countries such as India. Strain biodiversity represents a fundamental source of specific and also potential rare characteristics, which contributes to increase the economics of fermentation. This natural biodiversity represents an essential source for selection of strains possessing desirable characteristics. The use of molecular method like RAPD helps in elucidation of genetic diversity among the isolated yeast strains thereby establishing their evolutionary relationships. This will enable researchers in designing novel strains carrying all desirable characteristicsThesisItem Open Access BIODIVERSITY OF FLUORESCENT PSEUDOMONADS FROM THE RHIZOSPHERE SOILS OF REDGRAM AND MAIZE CROPS(ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, 2011) SUPRAJA, Y; SUBHASH REDDY, RIn the present investigation, fluorescent pseudomonads were isolated from the rhizosphere of maize and redgram crop plants, since the microorganisms that grow in the rhizosphere provide defense for roots against the attack by plant pathogens. Thirty bacterial isolates were obtained from the 50 rhizosphere soil samples. Out of which, fifteen isolates were identified as fluorescent Pseudomonads based on their fluorescence nature under UV-light on Pseudomonas agar. To test the antagonistic potential of isolated fluorescent pseudomonads they were screened against two common soil borne fungi viz., Fusarium moniliforme and Macrophomina phaseolina under in vitro conditions using dual culture plate technique. In this study, Pseudomonas (RPF-5, MPF-4, RPF-7, RPF-4, MPF-6 and MPF-2) isolates showed highest potential for antagonism against Fusarium moniliforme. Three Pseudomonas isolates (RPF-5, MPF-6 and RPF-8) were found as potential antagonists for Macrophomina phaseolina. The two isolates RPF-5 and MPF-6 had the capacity to inhibit the growth of two fungal pathogens. Under field conditions, in addition to biocontrol agents, farmers use agrochemicals like fungicides, insecticides and herbicides etc. to control plant diseases, pests and weeds. Because of this, the potential antagonistic PF (fluorescent Pseudomonas) isolates were tested for their compatibility with the commonly used agrochemicals. All the agrochemicals were found to be compatible with the ten PF (RPF-1, RPF-2, RPF-3, RPF-4, RPF-5, RPF-7, RPF-8, MPF-2, MPF-4 and MPF-6) isolates. To know the nature of the antagonistic property of PF isolates, tests were conducted for the production of siderophores and Hydrogen cyanide (HCN). They were also examined for the production of Indole acetic acid (IAA) and solubilization of phosphate. It was observed that all the isolates, which showed antagonism against fungal pathogens, were also found positive for IAA and siderophore production. All the isolates showed phosphate solubilization except RPF-7 and MPF-4. Only RPF-5 and RPF-8 were positive for HCN production. In order to know the genetic similarity of isolated fluorescent Pseudomonads, RAPD analysis was carried out by using twenty operan series (OPL) primers. In this study, all the isolates were proved cent per cent polymorphic based on polymorphic bands formed and dendrogram analysis. At 61% similarity level 6 clusters were distinguished with 15 isolates of fluorescent Pseudomonas isolates by RAPD using twenty primers (OPL series). In that the best isolates RPF-5, RPF-8 and MPF-6 were assimilated in to different clusters. RPF-5 formed a separate major subgroup with 59 per cent similarity coefficient. RPF-5 was best isolate in the present study for production of IAA, siderophore, HCN and phosphate solubilization. It exhibited antagonistic activity against Fusarium moniliforme and Macrophomina phaseolina. Another best isolate, MPF-6 belongs to 2nd cluster could produce Indole acetic acid, siderophores and phosphate solubilization ability. RPF-8, isolate formed 5th cluster. It showed similar qualities like RPF-5. Eventhough these three isolates showed similar qualities but variability is higher among these three isolates. It means that the genotypic and phenotypic diversity of P. fluorescens could be based on the geographic origin and functional differences in the genome of the isolates. At 55 per cent similarity coefficient all the fifteen isolates are genetically relatedThesisItem Open Access BIOGAS PRODUCTION FROM AGRICULTURAL WASTES AND FEASIBILITY STUDY TO ENRICH BIOGAS MANURES(ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, RAJENDRANAGAR, HYDERABAD, 2014) KAVYA, YERASI; VIJAYA GOPAL, AThe experiment was conducted during 2013-14 at Department of Agricultural Microbiology and Bioenergy, and Department of Soil Science & Agricultural Chemistry, College of Agriculture, Rajendranagar, ANGRAU, Hyderabad. Cow dung along with other agricultural wastes (press mud, poultry litter, kitchen wastes, maize stalks and fruit wastes) were used for the biogas production in lab scale. Along with the estimation of biogas production different parameters like Total Solids (TS) per cent, Total Volatile Solids (TVS) per cent, Volatile Fatty Acids (VFA), pH, Nitrogen (N), Phosphorous (P), Potassium (K), Organic carbon per cent, Biological Oxygen Demand (BOD), Chemical Oxygen Demand (COD), Electrical Conductivity (EC) and methane percentage was estimated. For each treatment 750 g of substrate and 1500 ml of water was used as inoculum mixture in 3 liters glass bottles. In T1 250 g cow dung + 500 g press mud + 1500 ml water, T2 250 g cow dung + 500g poultry litter + 1500 ml water, T3 250 g cow dung + 500 g kitchen wastes + 1500 ml water, T4 250 g cow dung + 500 g maize stalks + 1500 ml water, T5 250 g cow dung + 500 g fruit wastes + 1500 ml water and T6 750 g cow dung + 1500 ml water were used. The experiment was run for ten weeks and all the parameters were observed in the initial, 7th, 14th, 21st, 28th, 42nd, 56th day of gas production. At the end of tenth week the gas production was significantly more in T1 (Cow dung + Press mud) 9903.31 ml, compared toT6 (Cow dung alone) 8103.31ml, T2 (Cow dung + Poultry litter) 6079.98 ml, T3 (Cow dung + Kitchen waste) 4066.63 ml, T5 (Cow dung + Fruit waste) 3373.32 ml and less in T4 (Cow dung + Maize stalks) 3099.97 ml. By the end of the process the degradation of total solids (TS) per cent was significantly more in T6 (Cow dung alone) 57.16 per cent and less in T5 (Cow dung + Fruit waste) 32.55 per cent. The degradation of total volatile solids (TVS) per cent was significantly more in T6 (Cow dung alone) 37.41 per cent and less in T5 (Cow dung + Fruit waste) 14.32 per cent. pH in the substrate was estimated and it was significantly more in T1 (Cow dung + Press mud) 7.66 and less in T3 (Cow dung + Kitchen waste) 5.77. By the end of the process pH was more in T6 (Cow dung alone) 6.82 and less in T5 (Cow dung + Fruit waste) 3.67. The N per cent and P per cent in the process of anaerobic digestion was reduced while K per cent was increased. At the end of the process the reduction in BOD was observed and significantly more reduction was observed in T5 (Cow dung + Fruit waste) 58.74 per cent and less reduction in T4 (Cow dung + Maize stalks) 27.68 per cent. At the end of the process, decrease in COD was observed and was more in T6 (Cow dung alone) 52.15 per cent and less reduction in T4 (Cow dung + Maize stalks) 18.77 per cent. The second experiment on feasibility to enrich the biogas manures with beneficial microorganisms was carried at Department of Agricultural Microbiology and Bioenergy, College of Agriculture, Rajendranagar, ANGRAU, Hyderabad. The slurry samples from all the six treatments and three replications were collected dried in trays under the sun until the moisture was 50 per cent. The dried manure samples were divided into two and one part was sterilized and the other was used as the same i.e. unsterilized. The population of beneficial microorganisms in the dried manures was estimated and the population of Pseudomonas (46.0×103CFU g-1) and Azospirillum (24.0×103CFU g-1) was more in T6 (Cow dung alone), the population of Rhizobium was more in T1 (Cow dung+ press mud) 46.0×103CFU g-1 and the population of Azotobacter was more in T2 (Cow dung + Poultry litter) and T6 (Cow dung alone) 42.0×103CFU g-1. The beneficial microorganisms used in the study for enrichment were Rhizobium, Pseudomonas, Azotobacter and Azospirillum. The organisms were inoculated individually and also as consortia in to the manure samples of all the six treatments and the viability was monitored. In the unsterilized manure samples enriched individually with beneficial microorganisms the viability of Rhizobium was significantly more in T5 (Cow dung + Fruit waste), for Pseudomonas viability was significantly more in T3 (Cow dung + Kitchen waste), for Azotobacter viability was significantly more in T6 (Cow dung alone) and for Azospirillum viability was significantly more in T5 (Cow dung + Fruit waste). In the sterilized manure samples the viability of Rhizobium was significantly more in T3 (Cow dung + Kitchen waste), for Pseudomonas viability was significantly more in T3 (Cow dung + Kitchen waste), for Azotobacter viability was significantly more in T2 (Cow dung + Poultry litter) and for Azospirillum viability was significantly more in T1 (Cow dung + Press mud). The viability appears to be good upto the end of fourth week irrespective of treatments and also in unsterilized and sterilized biogas manure samples for Rhizobium, Pseudomonas and Azospirillum whereas for Azotobacter the viability appears to be good only upto the end of third week irrespective of treatments and also in unsterilized and sterilized biogas manures. In the unsterilized manure samples enriched with consortia of beneficial microorganisms the viability of Rhizobium was significantly more in T5 (Cow dung + Fruit waste), for Pseudomonas the viability was significantly more in T5 (Cow dung + Fruit waste), for Azotobacter viability was significantly more in T6 (Cow dung alone) and for Azospirillum viability was significantly more in T6 (Cow dung alone). In the sterilized manure samples the viability of Rhizobium was significantly more in T1 (Cow dung + Press mud) for Pseudomonas viability was significantly more in T5 (Cow dung + Fruit waste), for Azotobacter viability was significantly more in T1 (Cow dung + Press mud) and for Azospirillum viability was significantly more in T2 (Cow dung + Poultry litter). The viability appears to be good upto the end of third week irrespective of treatments and also in unsterilized and sterilized biogas manure samples for Rhizobium and Azospirillum whereas for Pseudomonas, the viability was good upto the end of fourth week irrespective of treatments in unsterilized biogas manures and upto the end of third week in sterilized manure samples. For Azotobacter, the viability appears to be good only upto the end of third week irrespective of treatments in unsterilized biogas manures and upto the end of fourth week in sterilized biogas manures.ThesisItem Open Access DEVELOPMENT AND EVALUATION OF PROMISING BACTERIAL CONSORTIA FOR IMPROVING GROWTH AND YIELD OF RICE(PROFFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2023-02-09) Srija, A.; Latha, P. C.Rice (Oryza sativa) is the most important cereal crop in the world, feeding more than 50% of the world's population. To meet the world's demand for rice, it is imperative to find environmentally sound ways that supplement the need for fertilizers. The use of microbial inoculants is a desirable option since they can complement the fertilizer use. Microbial formulations developed using two or more beneficial plant growth promoting rhizobacteria, colonize the rhizosphere, plant roots and aids in plant development. Therefore, inoculating the plants with microbial consortia produces positive results because microbial consortia can generally accomplish tasks by mutual synergism thus performing better than individual strains. The present study “Development and evaluation of promising bacterial consortia for improving growth and yield of rice” was carried out to identify a potential consortia for rice. A total of 32 (IIRRSS22-1 to 27, P1, R1, M1, O1 and P2) bacteria isolated from the rhizosphere of rice were screened and scored for plant growth-promoting traits (phosphate, potassium, zinc solubilization, siderophore production, indole acetic acid production and HCN production) under in vitro conditions. Three potential (P1, IIRRSS22-1 and IIRRSS22-6) isolates having highest score with regard to plant growth promoting traits were selected and the bacterial performance was evaluated further by conducting germination test. All the isolates were then screened for abiotic stress tolerance such as salinity, temperature and drought tolerance. Scores were assigned to the isolates based on their ability to grow at different NaCl concentrations, temperature of 45℃ and water potential of -0.73MPa and three isolates (IIRRSS22-3, R1 and IIRRSS22-7) with highest score for abiotic stress tolerance traits were selected for in vitro germination studies. Agrichemical compatibility of bacterial isolates with fertilizers (Urea, Single super phosphate and Muriate of potash), insecticides (Cartap, Ferterra, Thiamethaxom), herbicides (Pretilachlor, Bispyribac sodium) and fungicides (Carbendazim, Mancozeb) used for rice cultivation was assessed by disc diffusion method. Based on the tolerance xv exhibited by the bacterial isolates, IIRRSS22-5, IIRRSS22-2 and IIRRSS22-4 with the highest tolerance score were selected for further experimentation. Nine isolates, 3 from each category (plant growth promoting traits, abiotic stress tolerance and compatibility with agrichemicals) were grouped into consortia (Consortium-1, 2 and 3) containing 3 bacterial isolates in each. These bacterial combinations were evaluated for compatibility among each other by cross streak method. All the isolates were compatible with each other and these consortia combinations and individual bacteria were evaluated for germination under in vitro conditions with Telangana sona (RNR-15048) as test variety. Consortium-1 exhibited highest germination percentage, seedling length, vigour index I and II when compared with control, individual inoculations and other consortia (C-2 and C-3). Based on the in vitro germination assay, consortium-1 was selected for further evaluation under pot culture conditions with different treatments tested at 100% RDF except the control treatment. The pot culture experiments were taken up during rabi 2021- 22, the plant morphological traits and yield traits of rice were recorded. The root length, shoot length, leaf area and plant biomass at active tillering and at the harvest stage were found to be highest with treatment T14 (100% RDF + Bacterial consortia (Seed treatment + Seedling root dip). The yield parameters of rice such as tiller number, panicle length, filled grains per panicle, test weight and grain yield as affected by different treatments were found to be highest in treatment T14 (100% RDF + Bacterial consortia (Seed treatment + Seedling root dip). The soil available nutrients and the nutrient uptake by the plants were also highest in the treatment T14 (100% RDF + Bacterial consortia (Seed treatment + Seedling root dip). The molecular characterization of bacterial partners in C-1 by 16S rRNA gene sequencing revealed that the bacterial isolates were Pseudomonas stutzeri (P1), Stenotrophomonas sp. (IIRRSS22-3), Achromobacter sp (IIRRSS22-5). The isolates in consortium-2 and 3 were identified as Achromobacter insuavis (IIRRSS22-6), Rhizobium sp, (IIRRSS22-1), Stenotrophomonas maltophilia (IIRRSS22-2), Ochrobacter anthropi (IIRRSS22-4). Achromobacter xylosoxidans IIRRSS22-7. Based on the results obtained, the present study showed that the consortia (C-1) which consisted of rhizobacterial isolates (Pseudomonas stutzeri, Stenotrophomonas sp, Achromobacter sp) with multiple plant beneficial traits along with tolerance to eco physiological stresses has enhanced rice plant growth and yield parameters under pot culture conditions. This study also suggests that the use of plant-beneficial bacteria as a consortium results in better plant performance rather than as a single strain.ThesisItem Open Access EFFECT OF FYM AND VAM ON DROUGHT TOLERANCE OF SPINACH (Beta vulgaris) (PALAK)(ACHARYA N. G. RANGA AGRICULTURAL UNIVERSITY, 2012) TIRUPATHI RAO HANUMANTHU, HANUMANTHU; SUMATHI, SA poly bag experiment was conducted at Department of Agricultural Microbiology and Bioenergy, College of Agriculture, Rajendranagar, Hyderabad during 2011-12 to study the “Effect of FYM and VAM on drought tolerance of spinach (Beta vulgaris) (PALAK)”. The experiment was laid out in completely randomized design with 3 replications and 8 treatment combinations. Recommended dose of chemical fertilizers (100% RDF, 50% RDF), farm yard manure and mycorrhizae were applied to the soil according to the treatments imposed. All green variety of spinach seeds (20) were sown in poly bags and watered at two levels i.e alternate day and every 4th day. The crop was harvested around 40 days (1st picking) and 60 days (2nd picking) after sowing. At each picking, yield was noted, and fresh leaf samples were used for estimation of total chlorophyll content, proline content,vitamin C content and total carotene content. Dry powdered leaf samples were used for estimation of total antioxidant activity, total phenolic content, nitrogen, phosphorous and potassium uptake. The physico - chemical properties of soil and microbial population in the soil were estimated before sowing and after harvesting of the crop. The results revealed that among the treatments with alternate day irrigation, yield was highest in FYM+100%RDF followed by 100%RDF+FYM+VAM. Addition of FYM to the treatment VAM+100%RDF significantly increased the yield. Significantly highest dry weight and chlorophyll content were observed in FYM+100%RDF followed by 50%RDF+FYM+VAM and 100%RDF+FYM+VAM. The treatment FYM+100%RDF showed significantly maximum number of leaves followed by 50%RDF+FYM+VAM, 100%RDF+FYM+VAM. Highest vitamin C content and phenolic content were recorded in both 50%RDF+FYM+VAM and 100%RDF+FYM+VAM. The vitamin C content was significantly lowest in the treatment with VAM. Addition of VAM to the FYM and chemical fertilizers significantly increased the vitamin C content. Highest antioxidant activity and total carotene content were found in 100%RDF+FYM+VAM treatment. Highest proline content was recorded in FYM+100%RDF followed by FYM+VAM and lowest amount of proline was found in VAM treatment. Highest nitrogen, phosphorous and potassium uptake was observed in the treatment with 50%RDF+FYM+VAM followed by FYM+100%RDF. The uptake was significantly increased when FYM was added to the VAM+100%RDF treatment. Lowest uptake of N, P and K was found in VAM treatment. Maximum pH and high EC values were recorded in the treatment 100%RDF. The organic carbon content was highest in FYM and bulk density was found to be maximum with 100%RDF+FYM+VAM. Highest Nitrogen and Phosphorous content were observed in the treatment 100%RDF. Significantly lowest nitrogen and potassium content was observed in VAM treatment. Higher population of Bacteria, Actinomycetes, PSB and Fungi were found to be associated with the FYM treatment while Azotobacter population was maximum in FYM+VAM and VAM+100%RDF treatments. Among the treatments given irrigation at every 4th day, significantly higher yield and dry weight was found in 100%RDF+FYM+VAM which were immediately followed by 50%RDF+FYM+VAM. The results indicate that VAM as such did not influence the dry weight of the spinach crop. The influence of VAM was observed only in combination with FYM and chemical fertilizers. Highest number of leaves and chlorophyll content were recorded in 100%RDF+FYM+VAM and 50%RDF+FYM+VAM respectively. Highest vitamin C content, antioxidant activity and phenolic content were found in 100%RDF+FYM+VAM treatment. On addition of VAM to FYM+100%RDF treatment vitamin C content was significantly increased. Carotene content was highest in FYM+100%RDF followed by 100%RDF+FYM+VAM and 100%RDF. Higher proline content was recorded with 100%RDF+FYM+VAM treatment and lowest proline content was recorded in VAM. The uptake of nitrogen and phosphorous was highest in 100%RDF+FYM+VAM. The treatments with VAM, FYM and RDF helped in higher uptake of nitrogen under water stress. Phosphorous uptake recorded high in 100%RDF+FYM+VAM. Highest Potassium uptake was found in 50%RDF+FYM+VAM and 100%RDF+FYM+VAM. Lowest uptake of nitrogen, phosphorous and potassium were found in the treatments VAM, VAM+FYM and FYM respectively. The higher pH and EC were observed in100%RDF. Highest organic carbon content recorded in FYM, FYM+100%RDF and FYM+VAM treatments. Highest bulk density was observed in 100%RDF+FYM+VAM. Highest Nitrogen and Phosphorous content was observed in 100%RDF treatment. Potassium content was highest in VAM+100%RDF treatment. However lowest nitrogen, phosphorous and potassium content were observed in VAM. Highest number of bacteria was found in FYM+VAM. Actinomycetes were highest in number in the treatment FYM+VAM and lowest in 100%RDF. Highest number of PSB was observed in the treatment 50%RDF+FYM+VAM. Azotobacter population was highest in FYM+VAM followed by 50%RDF+FYM+VAM treatment and lowest in 100%RDF. Fungal population was highest in the treatment FYM and lowest in 100%RDF. The results obtained in the present experiment indicate that the FYM or VAM or RDF alone did not help in either higher yield or withstanding drought stress. Though FYM with RDF gave the best results than RDF alone, use of VAM with FYM and RDF gave the best results in terms of yield, antioxidant activity, vitamin C content both under normal and drought stress conditions. The % difference in yield was lowest in VAM treatments when compared between the two irrigation levels indicating that the VAM treatment influences yield under drought stress.ThesisItem Open Access EVALUATION OF CARRIER AND LIQUID BIOFERTILIZER USING DIFFERENT METHODS OF APPLICATIONS IN MUNGBEAN CROP(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY. HYDERABAD, 2016) BHAVYA, K; R. Subhash ReddyThe experiment conducted during 2015-16 at department of Agricultural Microbiology and Bioenergy, College of Agriculture, Rajendranagar, PJTSAU. Different carrier and liquid based biofertilizers are brought from different production units for evaluation of their quality. In case of carrier based biofertilizers, population of beneficial bacteria, pH, consistency, moisture content, level of contaminants were estimated. In case of liquid based biofertilizers, population of beneficial bacteria, pH, consistency and level of contaminants were estimated. The shelf life of biofertilzers were estimated using suitable media for viable count. Microbial population of beneficial bacteria was monitored in liquid and carrier based biofertilizers at monthly intervals. In liquid based biofertilizers viable count was constant for four months but a gradual decrease was observed in fifth and sixth months. Quality parameters of liquid based biofertilizers are good and constant for six months. In carrier based biofertilizers, the quality was too low, moisture content was high and count was decreased more. The field experiment was conducted following complete randomized design with control and 12 treatments. Treatments consisted of Control : 100% RDF, T 1 – T12 : 75% RDF +25% biofertilizer application, T 1 : CBF (seed treatment ) at the time of sowing, T2 : CBF (soil application) before sowing, T3 : CBF- liquid culture as soil application at the time of sowing, T4 : LBF - liquid culture (soil application) before sowing, T5 : LBF (seed treatment) at the time of sowing, T6 : LBF (soil application) before sowing, T7 : CBF (seed treatment at the time of sowing) + soil application at 40DAS, T8 : CBF (soil application before sowing) + soil application at 40 DAS, T9 : LBF (seed treatment at the time of sowing) + soil application at 40 DAS, T10 : LBF (soil application before sowing) + soil application at 40 DAS, T 11 : CBF-liquid culture(soil application at the time of sowing soil application at 40DAS), T 12 : LBF- liquid culture (Soil application before sowing) + soil application at 40 DAS.The growth parameters viz., plant height, root length, nodule number per plant, nodule dry weight (gm per plant), dry matter accumulation (gm per plant), were observed at different crop growth stages. Likewise observation on yield attributes viz., number of seeds per pod, test weight of seeds, seed yield were taken at harvest and nutrient uptake was estimated at harvest stage. Also available soil N, P and K were recorded at initial and at harvesting stage. Microbial population was recorded at initial, flowering and harvest stage. Application of carrier and liquid based biofertilizers in mungbean showed best result for the treatment T9 - Seed treatment with LBF at the time of sowing + soil application at 40 DAS. The treatment recorded maximum yield, number of seeds per pod and 100 seed weight than all other treatments. Treatment T10 - soil application with LBF at the time of sowing + soil application at 40 DAS was on par with T9. Highest nitrogen, phosphorous, potassium uptake, and higher available nitrogen, phosphorous, potassium in the soil were observed in the treatment T9 - LBF as seed treatment at the time of sowing + soil application at 40 DAS which was significantly higher than the biofertilizers applied through seedtreatment and soil application only once. At flowering and harvesting stage, treatment T9 - seed treatment with LBF at the time of sowing + soil application at 40 DAS recorded significantly higher bacterial, fungal, Rhizobium, Bacillus and actinomycetes population as compared to all other treatments. The significantly lowest bacterial, fungal, Rhizobium, Bacillus and actinomycetes population recorded with the treatment T3 - CBF Liquid culture as soil application at the time of sowing than all other treatments. .ThesisItem Open Access EVALUATION OF CARRIER BASED AND LIQUID BASED BIOFERTILIZERS AND THEIR APPLICATION METHODS IN GREENGRAM(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2018) SRAVANI, K; TRIVENI, SThe field experiment was conducted during kharif 2016 - 17 at Water Technology Centre fields, College farm with simple randomized block design which consisted 10 treatemnts and 3 replications. The laboratory work pertaining to this experiment was carried out at Department of Agriculutrual Microbiology & Bioenergy, College of Agriculture, Rajendranagar, PJTSAU, Hyderabad, Telangana. Evaluated the shelf life of CBBF and LBBF brough from different production centers by using specific media for Rhizobium – YEMA with congored and for PSB - pikovskayas agar. The significantly highest viable count with no contaminants were found in K.N. Biosciences produced liquid based biofertilizers i.e Rhizobium and PSB and they formed the maximum no. of cells in initial count was 8.38, 8.15 log no. of cells respectively. The plant growth parameters like plant hight and total plant dry matter production was significantly influenced by the seed inoculated with LBBF (Rhizobium and PSB) at 30, 45, 60 DAS and at harvest stage.The plant height was registered 27.40, 39.00, 46.33 and 47.07 cm respectively and plant dry matter production was noticed 8.60, 15.80, 22.30 and 18.30 g plant-1 at 30, 45, 60 DAS and at harvest respectively over control at all crop growth stages. The Seed inoculated with CBBF noticed significantly highest root length 12.63, 17.60, 22.83 and 23.50 cm at 30, 45, 60 DAS and at harvest stage of crop when, compared to other treatment combinations. The no. of root nodules and nodule dry weight was significantly influenced by the soil application of LBBF at 30 and 45 DAS. The significantly lowest no. of noules and their dry weight was recorded with treatment T10 (Control) i.e. only biofertlizers application. The total soil microbial population at flowering, harvesting stage and soil microbial biomass carbon, soil enzyme activities viz., soil dehydrogenase, acid and alkaline phosphatase and urease activity were significantly influenced by the drip fertigation with 100 % RDF and along with LBBF as biofertigation were registered maximum microbial population and their activity at crop harvest stage. Whereas, lowest was observed with control treatment. The maximum nutrient uptake (N P K) and their availability in soil was significantly recorded highest in treatment with T6 (100 % RDF and LBBF drip fertigation).The highest N P K uptakes values were 62.23, 24.10 and 51.33 kg ha-1 respectively at crop harvest stage. The highest yield & yield attributing characteristics of greengram like number of pods plant-1, number of seeds pod-1, test weight and seed yield (1019.50) kg ha-1, haulm yield (2543.50) kg ha-1 were noticed significantly highest by the application of liquid based biofertilizers along with 100 % RDF as fertigation at harvest satge. It was recorded significantly increased highest seed and haulm yield were (23.93 %) and (28.16 %) over control. The results showed that LBBF has significantly influenced the yield of greengram due to having more shelf life and when it is applied as biofertigation the microbial inoculants significantly influence the crop growth and yield of greengram during the crop period.ThesisItem Open Access EVALUATION OF EFFICIENCY AND PERSISTENCE OF DIFFERENT PHOSPHATE SOLUBILIZING BIOFERTILIZERS WITH MAIZE CROP(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY. HYDERABAD, 2016) VINOD BABU, S; TRIVENI, SThe beneficial microbes like phosphate solubilizing bacteria (PSB) are known to play an important role in supply phosphorous (P) to plants in a sustainable manner in P deficient soils. The mineral nutrition of plants mainly depends on soil P content that can be assimilated as a soluble phosphate. P is involved in all major metabolic processes in plants such as biosynthesis of macromolecules, energy transfer, photosynthesis, signal transduction and respiration. Keeping in view the importance of Phosphate solubilizing biofertilizers an experiment entitled “Evaluation of efficiency and persistence of different Phosphate solubilizing biofertilizers with maize crop” was conducted at Department of Agricultural Microbiology and Bioenergy, College of Agriculture, Rajendranagar, Hyderabad. Twenty four (24) phosphate solubilizing bacteria (i.e., sixteen Bacillus and eight Pseudomonas) isolated from Maize research station and college farm, Rajendranagar, PJTSAU, Telangana. The isolated bacterial isolates (24) were screnned for solubilization of insoluble forms of phosphorus, zinc and potassium. The solubilization zone for phosphate ranged from 15.50 mm to 6.10 mm. The isolate PSB 6 showed maximum solubilization zone of 15.50 mm. The solubilization efficiency (%) ranged from 258.33 % to 114.28 % which was maximum for PSB 6 (258.33 %). The solubilization zone for zinc ranged from 14.00 mm to 6.40 mm. The isolates both PSB 6 and PSB 19 showed maximum solubilization zone of 14.00 mm. The solubilization efficiency (%) ranged from 233.30 % to 110.34 % which was maximum for PSB 6 (233.30 %). The solubilization zone for potassium ranged from 14.00 mm to 6.00 mm. The isolate PSB 11 showed maximum solubilization zone of 14.00 mm. The solubilization efficiency (%) ranged from 280.00 % to 126.98 % which was maximum for PSB 11 (280.00 %). All the phosphate solubilizing bacterial isolates were checked for their purity and then studied for the morphological, cultural and biochemical characters. All the phosphate solubilizing bacterial isolates were subjected to further study to understand their Plant Growth Promoting Properties (PGPR) under in vitro conditions. Pure isolates were screened for IAA production, exopolysacharide production, siderophore production and HCN production. The best performed isolates were further tested for their compatibility with the fungi in vitro, for efficient biofilm formation. Persistency of Phosphate solubilizing biofertilizers in soil was recorded at different growth stages viz., vegetative, flowering and harvesting stages of crop. There was an increasing trend followed from the vegetative to flowering stages and also a gradual decrease was observed from flowering stage towards harvesting stage. Similar trend was observed for all the treatments studied. But significant counts were recorded in the treatment T7 (Carrier + Liquid + Biofilmed PSB biofertilizer) at vegetative (22.40x107cfu g-1), flowering (26.60x107cfu g-1) and at harvesting (22.03x107cfu g-1) respectively compared to all other treatments. High enzyme activity was observed at all the three growth stages for the T7 treatment, flowering stage being the highest for dehydrogenase (81.53 μg TPF d-1 g-1), acidic (123.00 μg p - nitro phenol g-1 h-1) and alkaline (146.20 μg p - nitro phenol g-1 h-1) phosphatase activity, urease (11.40 μg of NH4+-N 5g soil-1 2h-1) activity as a result of high persistency of PSB at this stage. This maximum enzyme activity increased plant growth, plant biomass and availability of nutrients. The major outcome of this study was the Carrier + Liquid + Biofilmed PSB biofertilizer treated Maize (Zea mays) plants showed best performance in all the biological parameters measured than untreated and individual culturesThesisItem Open Access EVALUATION OF QUALITY AND PERFORMANCE OF LIQUID AND CARRIER BASED BIOFERTILIZERS(ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, RAJENDRANAGAR, HYDERABAD, 2014) HIMA BINDU, JUTTU; SUBHASH REDDY, RThe experiment was conducted during 2013-14 at college farm, Department of Agricultural Microbiology and Bioenergy, and Department of Soil Science & Agricultural Chemistry, College of Agriculture, Rajendranagar, ANGRAU, Hyderabad. Different carrier and liquid based biofertilizers were brought from different firms for evaluation of their quality. In case of carrier based biofertilizers, population of beneficial bacteria, pH, consistency, moisture content, level of contaminants were estimated. In case of liquid biofertilizers, population of beneficial bacteria, pH, presence of contaminants were estimated. The shelf life and purity of liquid biofertilizers were tested using suitable media for viable count. Microbial population of beneficial bacteria was monitored in liquid and carrier based biofertilizers at monthly intervals. Liquid based biofertilizers were having more shelf life than carrier based biofertilizers. Microbial count in LBF was constant over a long period than CBF. In CBF, the microbial count was decreasing at a fast rate. Quality parameters of LBF are good and constant for 8 months as per the experiment. In CBF, the quality was too low and the moisture content is too high. The field experiment was conducted following randomized block design with 11 treatments and three replications. Treatments consisted of T1: 100 % RDF, T2: 75 % RDF + carrier based biofertilizers (seed treatment), T3: 75 % RDF + carrier based biofertilizers (soil application), T4: 75 % RDF + carrier based biofertilizers (soil application + FYM), T5: 75 % RDF + liquid based biofertilizers (seed treatment), T6: 75 % RDF + liquid based biofertilizers (soil application), T7: 75 % RDF + liquid based biofertilizers (soil application + FYM), T8: carrier based biofertilizers (seed treatment at sowing and soil application after 6 weeks), T9: carrier based biofertilizers (soil application at sowing and after 6 weeks), T10: liquid based biofertilizers (soil application at sowing and after 6 weeks), T11: liquid based biofertilizers (seed treatment at sowing and soil application after 6 weeks). The growth parameters viz., plant height, root length, nodule no. plant-1, nodule dry weight (g plant-1), dry matter accumulation (g plant-1), were recorded at different crop growth stages. Likewise observation on yield attributes viz., number of seeds pod-1, weight of 100 seeds, seed yield were taken at harvest and nutrient uptake was estimated at harvest stage. Also available soil nitrogen, phosphorus and potassium and microbial population were recorded at initial, flowering and harvest stage. Application of carrier and liquid based biofertilizers in soybean showed best result for the treatment T7 (75 % RDF + Liquid based biofertilizers soil application with FYM). The treatment recorded maximum yield, number of seed per pod-1 and 100 seed weight than all other treatments. Treatment receiving only liquid based biofertilizers and carrier based biofertilizers with FYM was on par with the treatment T7. Highest nitrogen, phosphorus and potassium uptake and higher available nitrogen, phosphorus, potassium in the soil were observed in the treatment receiving 75 % RDF + Liquid based biofertilizers soil application with FYM which was significantly higher than the treatments of only seed treatment or only soil application. At harvest, treatment T7 (75 % RDF + Liquid based biofertilizers soil application with FYM) recorded significantly higher bacterial, fungal, actinomycetes, Rhizobium and Pseudomonas population as compared to the other treatments. The significantly lowest bacterial, fungal, actinomycetes, Rhizobium and Pseudomonas population was found in the treatment T1 (100 % RDF).ThesisItem Open Access FERMENTATIVE PRODUCTION OF BIO-ETHANOL FROM OVER RIPENED SAPOTA FRUITS(ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY RAJENDRANAGAR, HYDERABAD, 2010) YELPALE MANOJ MARUTI; SUBHASH REDDY, RFossil fuels depletion is one of the major issues, the world is facing at present. The use of fossil fuels has an adverse impact on environment due to the oxidation of CO2, SO2 and NO2 emissions. To overcome such problems, alternative fuels such as bioethanol and biodiesel can be used as replacement for fossil fuels. Bioethanol can be derived from biomass with different materials such as corn, sugarcane, wood and fruits wastes that are renewable and sustainable resources, which are easily accessible and reliable, and can help in avoiding environmental pollution due to dumping of wastes. Currently, there is a growing interest for ecologically sustainable bio-fuels, such as bio-ethanol produced by fermentation from renewable sources as fuel or fuel additive. The present investigation is carried out with the objective of isolating ethanologenic yeasts from different sources, and alcohol production from over ripened sapota fruits. Ten samples were obtained from different sources and pure isolates were identified as yeast cultures based on their morphological, cultural characters and alcohol producing ability. All isolates had alcohol producing ability but maximum alcohol production was observed with Saccharomyces cerevisiae MTCC-172 (67.85g/L) and isolate AMY-2 (67.80g/L) while isolate AMY-7 produced the least (29.80g/L). Over ripened sapota fruit pulp was subjected to pectinase treatment to increase the flowability of pulp. Different pHs, temperatures and yeast cultures were used for optimizing the fermentation conditions for bioethanol production from over ripened sapota fruits. It was found that pH (5.0) was suitable for higher alcohol production than lower pH (4.0 & 4.5) with both the cultures. Higher alcohol production was observed at controlled temperature (260C) with both cultures at different pH conditions. It was also found that alcohol fermentation of over ripened sapota fruits could be completed within 24 h. Alcohol production was found to be higher (67.00g/L) with both cultures Saccharomyces cerevisiae MTCC-172 and culture AMY-2 (58.19g/L) when the fermentation conditions pH 5.0 and 260C temperature were used with over ripened sapota pulp as substrate. Based on present study, it is established that over ripened sapota fruits can be used effectively for fermentative production of alcohol with pH 5.0 and temperature 260C as fermentation conditions in 24 h.ThesisItem Open Access IDENTIFICATION OF EFFECTIVE ISOLATE OF PSEUDOMONAS FROM THE SOILS OF SOUTHERN TELANGANA ZONE AGAINST CASTOR WILT PATHOGEN(ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY RAJENDRANAGAR, HYDERABAD, 2009) JAYAMMA, P.; SUBHASH REDDYWilt of castor caused by Fusarium oxysporum f.sp. ricini which was earlier considered as a disease of minor importance, now it became a major disease and an impediment in the cultivation of the castor. Continuous use of agrochemicals in pest and disease control leads to environment, soil and water pollution, where as biological control of diseases is long lasting, less expensive, eco-friendly and harmless to non target organisms. Keeping in view the benefits of biological control of plant pathogens the present investigation is carried out. Symptoms of the disease on seedlings showed characteristic yellowing and bronzing of cotyledonary leaves associated with degenerated root system. Symptoms on grownup plants manifested as stunting of the plant, yellowing of leaves, production of black streaks on stems and discolouration of roots. The fungus associated with the disease was isolated and identified as Fusarium oxysporum f.sp. ricini and its pathogenicity was proved under greenhouse conditions. Microorganisms are known to grow in the rhizosphere and provide frontline defence for roots against the attack by plant pathogens. In the present study, fluorescent Pseudomonads were isolated from the rhizosphere of crop plants like sorghum, maize, groundnut, chilli, tomato, castor and also from various forest plants. Twenty seven isolates were obtained from the 70 rhizosphere soil samples. Out of which, eighteen isolates were identified as fluorescent Pseudomonads based on their fluorescence nature under UV- light on Pseudomonas agar. Isolated fluorescent Pseudomonads were screened against Fusarium oxysporum f.sp. ricini under in vitro conditions using dual culture plate technique to test the antagonistic potential. Three fluorescent Pseudomonas isolates (STPF-11, STPF-14 and STPF-18) were found to be potential antagonists against the tested fungal pathogen. Under field conditions, agrochemicals like fungicides, insecticides and herbicides etc. are used to control plant diseases and pests. In addition to agrochemicals biocontrol agents are also used by farmers for controlling some diseases. Because of this, the potential antagonistic PF (fluorescent Pseudomonads) isolates were tested for their compatibility with the commonly used agrochemicals. Among the different agrochemicals tested carbendazim, chlorpyriphos and pendimethalin were found to be more compatible with PF isolates. Disease management strategy was developed for wilt of castor under greenhouse conditions. Among the treatments, treatment T3 (seed treatment with STPF-14 isolate + soil application of Farm Yard Manure (FYM) incubated STPF -14 isolate) was highly effective with least per cent disease incidence of 26.56 per cent at 45 days after sowing. This treatment not only reduced the disease incidence to a maximum extent, but also recorded maximum plant height (27.32 cm) root length (6.36 cm) and maximum fresh (21.32 g) and maximum dry weight (5.19 g) of castor when compared to other treatments.ThesisItem Open Access IMPACT OF BIOFERTILIZERS ON NUTRIENT CONTENT AND ANTIOXIDANT ACTIVITY OF SPINACH BEET (Beta vulgaris)(ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, 2011) REVATHI, K; SUMATHI, SA poly bag experiment was conducted at Department of Agricultural Microbiology and Bioenergy and Inter faculty Department of Biochemistry, College of Agriculture, Rajendranagar, Hyderabad during 2010-11 to study the Impact of biofertilizers on nutrient content and antioxidant activity of spinach beet (Beta vulgaris)”. The experiment was laid out in completely randomized design with 3 replications and 12 treatment combinations. Recommended dose of chemical fertilizers (100% RDF, 50% RDF), vermicompost and biofertilizers (nitrogen biofertilizer (NBF): Azotobacter and phosphate solubilizing biofertilizer (PSBF): Bacillus) were applied to the soil according to the treatments imposed. All green variety of spinach beet seeds (20) were sown in poly bags and watered regularly. The crop was harvested at 30 (1st picking), 45 (2nd picking) and 60 (3rd picking) days after sowing. At each picking, yield was noted, and fresh leaf samples were used for estimation of vitamin C content and total carotene content. Dry powdered leaf samples were used for estimation of total antioxidant activity, total phenols and mineral content. The physico - chemical properties of soil and microbial population in the soil were estimated before sowing and after harvesting of the crop. The shelf life of the leaf samples was assessed both at refrigeration and ambient temperature in 150 gauge polythene covers. The results revealed that the treatment receiving 50% RDF + NBF + PSBF recorded higher yield followed by 100% RDF, 50% RDF + NBF, 50% RDF + PSBF and vermicompost + 50% RDF. Dry weight was significantly higher on application with 50% RDF + NBF + PSBF which was on par with 100% RDF, 50% RDF + PSBF, 50% RDF + NBF and vermicompost with 50% RDF. The application of 50% RDF + NBF + PSBF resulted in higher vitamin C content over the other treatments at 30, 45 and 60 days after sowing. Low vitamin C content was recorded with vermicompost + NBF application and with PSBF application. Total carotene content was significantly higher with application of 50% RDF + NBF + PSBF at 30, 45 and 60 DAS over all other treatments. Calcium content was found to be significantly higher with the application of 50% RDF + NBF + PSBF followed by vermicompost with biofertilizers alone or combined and with 50% RDF. The lowest calcium levels were observed with only vermicompost treatment followed by 100% RDF. Iron content was found to be significantly higher with the application of 50% RDF + NBF + PSBF followed by vermicompost + 50% RDF and with two biofertilizers i.e., NBF and PSBF. Among all treatments total antioxidants were increased significantly with 50% RDF + NBF + PSBF which was on par with vermicompost + 50% RDF treatment. There was significant difference among 30, 45 and 60 days after sowing in the total antioxidant content. Significantly maximum total phenolic content was recorded in 50% RDF + NBF + PSBF followed by vermicompost + NBF + PSBF, vermicompost + 50% RDF and vermicompost + NBF. The lowest phenolic content in spinach beet leaves was found with vermicompost treatment and all other treatments showed significantly higher total phenol content. Shelf life was higher in the 50% RDF + NBF + PSBF treatment followed by vermicompost + NBF + PSBF, NBF + PSBF and vermicompost + 50% RDF both at refrigeration and room temperature in polythene cover. The soil pH was decreased with application of organic fertilizers and biofertilizers and EC was increased. The organic carbon content, nitrogen, phosphorus and potassium levels in the soil increased with 50% RDF + NBF + PSBF combination. Significatly higher soil microbial population was recorded with 50% RDF + NBF + PSBF treatment. Azotobacter and Bacillus population were higher in the treatments with biofertilizers alone or in combination, and with vermicompost. Lowest soil microbial population was recorded in 100% RDF. 50% RDF with NBF and PSBF was found to be the best among the treatments used in the present study as it resulted in the higher yield, vitamin C, total carotene, nutrient contents and antioxidant activity.ThesisItem Open Access INFLUENCE OF MICROBIAL CULTURES AND ORGANIC MANURE ON LEAFY VEGETABLES GROWN IN POLLUTED SOILS AND POLLUTED WATER(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2018) NISSI PAUL. MARAPATLA; Triveni, TThe experiment was conducted during 2016 -17 at Department of Agricultural Microbiology and Bioenergy, College of Agriculture, Rajendranagar, PJTSAU, Hyderabad. Heavy metal pollution is currently a major environmental problem because metal ions persist in the environment due to their non-degradable nature. The toxicity and bioaccumulation tendency of heavy metals in the environment is a serious threat to the health of living organisms. Bioremediation is a state-of-the-art technique used for heavy metal removal or recovery from polluted environments. The technique utilizes inherent biological mechanisms to eradicate hazardous contaminants using microorganisms and plants, or their products. Microorganisms are omnipresent that dominate in heavy metalcontaminated soil and can easily convert heavy metals into non-toxic forms. A total of 28 isolates were collected from different polluted sites of Hyderabad where green leafy vegetables were grown. The isolates were screened for their PGPR properties, the isolates were also tested for their heavy metal tolerance. The best five potential heavy metal tolerant PGP bacteria were studied for their molecular identification and were identified by 16S rRNA gene sequencing. BLAST search through NCBI revealed 97 % similarity of SfS-5 with Bacillus megaterium, 98 % similarity of AfS-8 with Pseudomonas aeruginosa, 98 % similarity of SfS-18 with Rhizobium meliloti, 99 % similarity of SfS-24 with Azotobacter chroococcum, 97 % similarity of SfW-28 with Azotobacter chroococcum. An experiment entitled “Influence of microbial cultures and organic manure on leafy vegetables grown in polluted soil and polluted water” was conducted in pot culture with 14 treatments and three replications during Rabi, 2016 and Rabi, 2017. The crops grown were spinach and coriander and seed treated with nitrogen fixer (SfS -24) and phosphate solubilizer (AfS - 8). The treatments imposed were T1: (Control- without fertilizers), T2: (Soil + Water + Biofertilisers), T3: (Soil + water + OM), T4: (Soil + water + RDF), T5: (Soil + water + Biofertilisers + OM), T6: (Polluted Soil+ water + Biofertilisers), T7: (Polluted Soil+ water + OM), T8: (Polluted Soil+ water + RDF), T9: (Polluted Soil+ water + Biofertilisers + OM), T10: (Soil + Polluted water + Biofertilisers), T11: (Soil + Polluted water + OM), T12: (Soil + Polluted water + RDF), T13: (Soil + Polluted water + Biofertilisers+ OM), T14: (Sterilized soil - control). The influence of various inorganic fertilizers, organic fertilizers and biofertilizers on plant height, root length, fresh weight and dry weight, available NPK, soil enzyme, soil microbial population was recorded at 20, 40, 60 DAS of the crop. The number of leaves and yield were determined. The concentrations of macro and micronutrients in plant at harvest were estimated. At the time of Harvest, highest plant growth parameters, dry matter accumulation, number of branches per plant, number leaves per plant yield were recorded in the treatment which received 100 % RDF (T8: Polluted Soil+ water + RDF). Plant height at 60 DAS was found highest in T8 with 30.4 cm in spinach and 45.4 cm in coriander; fresh weight at 60 DAS was registered significantly highest in T8 with 30.4 g in spinach and 11.0 g in coriander. Soil biological quality was high in treatments supplied with biofertilizer along with organic manure application. Bacterial population count was enumerated and found to be significantly highest in T9: Polluted soil + water + Biofertilisers + OM with 101.1 × 107 CFU g -1 of soil with spinach and 92.1 × 107 CFU g -1 of soil with coriander at 60 DAS , Similarly, soil enzyme activity (dehydrogenase, urease and phosphatase) was significantly higher in treatment T9 for both the crops at the time of harvest. Higher heavy metal concentration in the soil was recorded in pots irrigated with polluted water (T10, T11, T12 and T13) with increasing concentration of available NPK. The heavy metal concentration was recorded significantly higher in T12: Soil + Polluted water + RDF (Ni-4.084, Cd-1.573, Co-1.789 and Pb-13.850 mg kg-1 ) in spinach grown soil, whereas, in coriander grown soil was (Ni-3.72, Cd-2.143, Co-0.830 and Pb-9.65 mg kg-1 ). With increase in soil heavy metal content, the accumulation in plant also increased but with decrease in soil microbial population and enzyme activity. Significantly lower levels of heavy metal concentration in both plant shoot and soil was observed in the treatments receiving biofertlisers and organic manures, among all T9 is the treatment showed highest removal even though its initial heavy metal concentration was high. Based on the results obtained in the present study, it is observed that the consortia of heavy metal tolerant PGPR bacteria with soil application of FYM can serve as cost effective, ecofriendly bioremediating source in heavy metal polluted urban areas.ThesisItem Open Access INFLUENCE OF MICROBIAL CULTURES ON BIOLOGICAL QUALITY OF POLLUTED SOILS AND SPINACH YIELD(ACHARYA N.G. RANGA AGRICULTURAL UNIVERSITY, RAJENDRANAGAR, HYDERABAD, 2014) UMA RAJASHEKHAR, ALAVALA; SUBHASH REDDY, RThe experiment was conducted during 2012-13 at Department of Agricultural Microbiology and Bioenergy, College of Agriculture, Rajendranagar, Professor Jayashankar Telangana State Agricultural University, Hyderabad. The treatments for poly bag experiment were fixed and twelve treatments each treatment with three replications were designed. The cleaned poly bags were filled with 8 kg of soil and this soil was mixed with chemical fertilizers (0.14: 0.24: 0.37 g poly bag-1 NPK), farm yard manure (78.75 g poly bag-1) and Vesicular Arbuscular Mycorrhizae (100 to150 g of infected propagules poly bag-1) according to the treatments which were neatly arranged in the net house. The poly bag experiment was conducted following complete randomized block design with 12 treatments and three replications. Treatments consisted of T1 SF Soil + FYM @ 12 T/ha, T2: SF Soil + FYM + VAM + Pseudomonas, T3: SF Soil + RDF, T4: SF Soil + RDF + FYM + VAM + Pseudomonas in polluted soil with supply of fresh water T5: Soil + FYM+ Pseudomonas, T6: Soil + FYM + VAM + Pseudomonas, T7: Soil + RDF , T8: Soil + RDF + FYM + VAM + Pseudomonas in unpolluted soil with supply of fresh water, T9: Soil + FYM, T10: Soil + FYM + VAM + Pseudomonas, T11: Soil + RDF, T12: Soil + RDF + FYM + VAM + Pseudomonas in unpolluted soil with supply of polluted water. Microbial population and enzyme activity of enzymes urease, dehydrgenase, acid and alkaline phosphatases were recorded at initial, 30 and 60 DAS. Available soil nitrogen, phosphorus and potassium were recorded at initial and 60 DAS. Dry matter accumulation i.e. fresh and dry weight (g plant-1) were recorded at 30 and 60 DAS. Microbial biomass carbon and leaf area measured at 60 DAS. The Ni, Co, Cd in stem & leaf portion of spinach plants analyzed by Atomic absorption spectrophotometer. The water samples were collected and analysed for BOD and COD. Effect of microbial cultures on microbial population at 30 and 60 DAS (107CFU g-1 of soil in polluted and unpolluted soils of spinach beet showed best result for the treatment T9: Soil + FYM and T12: Soil + RDF + FYM + VAM + Pseudomonas. Effect of microbial cultures on enzymes urease, dehydrogenase, acid and alkaline phosphatases were recorded maximum enzyme activity in Treatment T8: Soil + RDF + FYM + VAM + Pseudomonas in unpolluted soil with supply of fresh water than all other treatments. Higher available nitrogen, phosphorus, potassium in the soil were observed in the treatment receiving T3: SF Soil + RDF which was significantly higher than the treatments. Highest leaf area and fresh and dry weight of leaf per plant were recorded in treatment T8: Soil + RDF + FYM + VAM + Pseudomonas in unpolluted soil with supply of fresh water. Heavy metal content in soil, leaf and shoot were highest for the treatments receiving SF Soil + RDF + FYM + VAM + Psuedomonas in polluted and unpolluted soils.ThesisItem Open Access INFLUENCE OF MULTIPLE APPLICATION OF MICROBIAL INOCULANTS ON YIELD OF BLACKGRAM(ACHARYA N. G. RANGA AGRICULTURAL UNIVERSITY, 2013) NAVEEN KUMAR, REDDI; SUBHASH REDDY, R.The experiment was conducted during 2012-13 at college farm, Department of Agricultural Microbiology and Bioenergy, and Department of Soil Science & Agricultural Chemistry, College of Agriculture, Rajendranagar, ANGRAU, Hyderabad. The field experiment was conducted following randomized block design with 9 treatments and three replications. Treatments consisted of T1 - 100% RDF, T2 - Uninoculated control, T3 - Only seed coating with inoculant, T4 - Only soil application with inoculant, T5 - Seed coating + soil application of inoculant with FYM, T6 - Seed coating + soil application of inoculant with FYM at 25 DAS, T7 - Soil application + soil application of inoculant with FYM at 25 DAS, T8 - Seed coating + soil application with FYM + soil application of inoculant with FYM at 25 DAS, T9 - Only soil application at 25 DAS. The growth parameters viz., plant height, root length, nodule no. plant-1, nodule dry wt. (g plant- 1), dry matter accumulation (g plant-1), were recorded at different crop growth stages. Likewise observation on yield attributes viz., number of seeds pod-1, weight of 100 Seeds, seed yield were taken at harvest. The protein % in seed and nutrient uptake were estimated at harvest stage. Also available soil nitrogen, phosphorus and potassium and microbial population were recorded at initial, flowering and harvest stage. At 25 DAS and 50 DAS, treatment T9 (only soil application at 25 DAS) recorded significantly higher plant height, whereas treatment T8 (seed coating + soil application with FYM + soil application of inoculant with FYM at 25 DAS) recorded significantly maximum root length as compared to all other treatments. At flowering stage treatment T8 (seed coating + soil application with FYM + soil application of inoculant with FYM at 25 DAS) recorded significantly higher number of root nodule and nodule dry weight and also total dry matter production and seed protein content at harvest stage compared to all other treatments. Multiple application of microbial inoculants i.e seed treatment + soil application of inoculants with FYM + at 25 DAS soil application of inoculants with FYM recorded maximum yield, number of seed per pod-1 and 100 seed weight than all other treatments. Highest nitrogen, phosphorus and potassium content and uptake, and higher available nitrogen, phosphorus, potassium in the soil were observed in the treatment receiving seed treatment + soil application of inoculants with FYM + at 25 DAS soil application of inoculants with FYM which was significantly higher than the inoculants applied through only seed treatment or only soil application or seed treatment + soil application only once. At harvest, treatment T8 (seed coating + soil application with FYM + soil application of inoculant with FYM at 25 DAS) recorded significantly higher bacterial, fungal, actinomycetes, Rhizobium and Pseudomonas population as compared to the other treatments. The significantly lowest bacterial, fungal, actinomycetes, Rhizobium and Pseudomonas population was found in the treatment T2 (un-inoculated control).ThesisItem Open Access ISOLATION AND CHARACTERIZATION OF ENDOPHYTIC BACTERIA FROM RICE ROOTS(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2018) HEMA, G; TRIVENI, SRice (Oryza sativa L) is the world’s most important staple food for two-thirds of the human population, especially in Asia. In India, 65 % of the population depends on rice as their staple food. Increases in the demand for rice, as a result of increase in population, creates the need to improve rice productivity. Application of plant growth promoting associative bacteria which can be a potential option for enhancing growth and yield of rice in sustainable manner. Plant growth promoting endophytic bacteria (PGPEB) are those bacteria living within plant tissues without causing any visible symptoms and increase plant growth by production of various plant growth hormones, phosphate solubilizing activity, nitrogen fixation and biocontrol activity. Present investigation was carried out to isolate and characterize PGPEB of rice. A total of 149 endophytic bacterial isolates were isolated from four rice genotypes collected from IIRR, Rajendranagar, Telangana. The isolates were purified, colony characters were recorded. The genotype BPT 5204 supported significantly higher mean endophytic bacterial population of 5.34 log CFUg-1 root among the genotypes while the lowest population was observed in IRGC 1220 (4.95 log CFUg-1root). All the 149 isolates were screened for nine PGP activities under in vitro conditions. No isolates among the 149 isolates produced hydrogen cyanide (HCN), which is proven biocontrol activity. Results revealed that among 149 endophytic isolates, 81.20 % isolates produced IAA, 34.89 % isolates solubilized phosphorous, 67.78 % isolates solubilized zinc, 40.93 % isolates solubilized potassium, 44.29 % produced siderophore, 44.29 % isolates were antagonistic against Rhizoctonia solani and 52.34 % isolates were antagonistic against Pyricularia oryzae. One isolate from each media having highest PGP activity score were selected for further evaluation through germination and seedling vigour studies. Compatibility test between bacterial isolates revealed that all the isolates were compatible against each other without inhibiting the growth of other endophytic isolates. Nine endophytic bacterial isolates which increased the vigour index of plants when compared to un-inoculated control were selected for pot culture studies. The nine isolates BN5, IN6, BT1, ST11, IT9, BR1, BRen1, BM3, MA1 were further evaluated as a consortia under pot culture studies. Bacterial consortium consisting nine best isolates (2.8 × 108 CFU ml-1) was used to treat rice seeds (MTU 1010) for pot culture study. Endophytic bacterial consortia significantly improved rice growth parameters like germination, plant height, root length, seedling biomass and number of tillers per plant. The plant growth parameters at 90 DAT were significantly higher in the treatment consisting of bacterial consortium + organic fertilizer + 75% RDF when compared to untreated control demonstrating the ability of saving in fertilizer application. The nine promising isolates were characterized morphologically and biochemically (IMVIC tests, oxidase test, catalase test, carbohydrate utilization test, starch hydrolysis and gelatin liquefaction) and by molecular characterization through sequencing of 16S rRNA gene. Based on the sequencing, the endophytic isolates were identified as Bacillus altitudinis (BN5), Bacillus altitudinis (IN6), Agrobacterium fabrum (BT1), Enterobacter cloacae (ST11), Agrobacterium fabrum (IT9), Bacillus altitudinis (BR1), Agrobacterium fabrum (BM3) and Halobacillus sp., (MA1). The isolate BRen1 was unidentified because of lack of BLAST hits and could be a novel bacterium. Based on the results obtained in the present study, the consortia of endophytic bacteria with multiple plant beneficial activities when formulated into a bioinoculant can serve as cost effective, ecofriendly bioproduct that can also help in minimising chemical application in rice production.ThesisItem Open Access ISOLATION AND CHARACTERIZATION OF Pseudomonas fluorescens FROM RICE FIELDS FOR THE BIOLOGICAL CONTROL OF Rhizoctonia solani CAUSING SHEATH BLIGHT IN RICE(PROFESSOR JAYASHANKAR TELANGANA STATE AGRICULTURAL UNIVERSITY, 2015) BIYYANI SUMAN; VIJAYA GOPAL DEPARTMENT, ASheath blight is a devastating disease caused by Rhizoctonia solani Kuhn in rice crop. Biocontrol agents have great demand now-a-days as they are replacing chemical pesticides to a large extent as they are cost effective, ecofriendly and easily available. Pseudomonas fluorescens is one among them which not only enhances the plant growth but also controls the fungal pathogens by production of anti fungal metabolites. Present investigation is focussed towards isolation of fluorescent Pseudomonas from rice rhizosphere and in vitro, in vivo evaluation of antagonistic activity of these isolated fluorescent Pseudomonas against Rhizoctonia solani causing sheath blight in rice crop. Thirty Pseudomonas fluorescens isolates were isolated from rice rhizospheric soils of twenty two villages of Parigi and Doma mandals of Rangareddy district, Telangana as the district has severe sheath blight incidence. Latitude, longitude, elevation, GPS numbers of the place of collection of soil samples are recorded and point map has been generated by using Arc GIS software and rectification of the toposheets was done by Erdas image 9.3 software. The isolates are purified by observing under UV light and they were culturally, morphologically and biochemically identified as Pseudomonas fluorescens according to Bergey’s manual of Determinative Bacteriology. Biochemical characterization revealed that all the isolates were positive for catalase, oxidase, citrate utilization, gelatine liquefaction, denitrification and negative for indole tests. These isolates were screened in vitro for Plant growth promoting attributes like phosphate solubilizatin, siderophore, IAA, ammonia and HCN production. Results revealed that 76.67% isolates solubilized phosphorous and produced siderophores. 93.33% isolates produced IAA and all the thirty isolates i.e., 100% produced ammonia and HCN. All the isolates were further screened in vitro for antagonistic activity against the fungal pathogen Rhizoctonia solani causing sheath blight and found that all isolates inhibited the fungal pathogen and highest inhibition was found with the organism DMP1 (53.43%). Pot culture experiment was conducted for in vivo evaluation of Pseudomonas fluorescens against the challenge inoculated rice crop by using various methods of inoculation. Results revealed that seed treatment method of application of Pseudomonas fluorescens with the isolate DMP1 effectively controlled and reduced the disease severity of 42.56% at 60 DAT, 41.12% at 70 DAT and 40.81% at 80 DAT caused by the Rhizoctonia solani compared with the other methods of inoculation. Seed treated plants especially with the organism DMP1 not only controlled the disease but improved the growth parameters like plant height (56 cm, 74 cm and 82.6 cm at 30,60 and 90 DAT respectively), Leaf area index (4.71 at 60 DAT), chlorophyll content (39.60 and 48.17 at 30 and 60 DAT ) and yield parameters like number of tillers (13.7, 17.3 and 17.3 at 30,60 and 90 DAT), panicles (13.7 at 90 DAT) and grain yield per plant (32.69 gms) followed by root dipping and foliar method of application of biocontrol agents. When the methods of inoculation are compared seed treatment method improved the growth and yield parameters in the challenge inoculated rice crop. When the challenge inoculated plants are scored for the disease, it was observed that the seed treated method of inoculation to the rice showed resistance towards the disease with a score of 1.2.