Using glucose as carbon source and mustard cake and yeast extract as nitrogen sources bacterial isolate Pseudomonas B2 exhibited a maximum PHB recovery of 0.620 (in terms of O.D.) and PHB weight of 0.27g/L in 96 h. To determine the possibility of growth potential of Pseudomonas spp., it was grown in different carbon sources like fructose, glucose, maltose, mannitol etc. and it was found that glucose yielded good growth and PHB production. In order to incorporate cost effective nitrogen and carbon source, mustard cake and cotton cake as nitrogen source and molasses as carbon were used in medium. Statistical media optimization design was used to optimize the culture conditions for maximizing the PHB production. A maximum of 0.37 g/L of PHB and 0.746 (O.D.) PHB recoveries were obtained using optimized concentrations. Batch kinetics can be used for model development, which will make possible simulation of nutrient limited cultivation(s) for over accumulation of PHB. FTIR studies confirmed the presence of PHB.
Conjugation between microsymbiont was used to create genetic variations in rhizobia for diverse rabi legumes (chickpea, pea and lentil) with better characteristics in nodulation and nitrogen fixation process. Ten antibiotics were used as selectable markers for the screening of twenty four bacterial strains to be used in mating experiments for obtaining transconjugants. All bacterial strains were sensitive to gentamycin and resistant to streptomycin, kanamycin and sulphanilamide. Total five fusants were obtained from each rhizobial cross combination with the help of electro-poration. Modified transconjugants, rhizobial strains had promiscuous infection with 50-122% more nodules showed significant increase in shoot fresh weight, dry weight and total nitrogen content in chickpea, pea and lentil plants. Electrofusants rhizobial strains improved shoot nitrogen content up to 67% in lentil and 54% in pea and chickpea plants. The amount of nitrogen fixed in chickpea was highest (3.71gm) by transconjugants DP-C6-HLN followed by DP-C6-HP14 (3.56gm). Transconjugants DP-HP14-HLN fixed the highest amount of nitrogen (3.92gm) in pea and 4.06gm in lentil plants. Plasmids were also analyzed in order to characterize their role in the evolution of rhizobial symbionts and their involvement in symbiotic behaviour. The developed Rhizobium strains with improved symbiotic association and ability to infect across strict specificity for host legumes would be of great help for the farming community at large.
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