Isolation and Evaluation of Nitrogen Fixing Bacteria in Tea Soils for the Production of Biofertilizer

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Date
2013
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Uva Wellassa University of Sri Lanka
Abstract
The field practices are very important and soil is very significant factor that consider in the agricultural crop production and management. Though each and every field practices are important, the soil fertility management is most significant one. Synthetic and organic fertilizers generally used to provide nutrient elements. Nitrogen is an essential nutrient component that has several beneficial effects on the crop growth. And also, nitrogen is a limiting nutrient which can be identify in the soil in cultivable lands for growth and yield of crops. Several kinds of microorganisms engage with Biological Nitrogen Fixation, Azospirillum bacteria is an associative micro aerophilic nitrogen fixer commonly found in association with the rhizosphere and roots of a variety of plants including cereals and grasses. They have the ability to produce usable form of nitrogen for the plants when they functioning in the soil biosphere. The present study was aimed to isolate Azospirillum spp.from tea soils, root parts of rehabilitation grasses in tea plantations and maize and study their nitrogen fixing abilities aiming at applying the Azospirillum as Biofertilizer in Tea lands. Methodology This research was carried out at the Microbiology laboratory of Uva Wellassa University, Soil samples and parts of root samples of Mana (Cymbopogan confertiflorus), Guatamala (Tripsacum laxum), Guinea (Panicum maximum) grasses and Maize (Zea mays) were collected from the university premises, farmer fields and tea estates closer to Uva Wellassa University, Badulla. Sample collection was done from well grown, healthy plants. All samples were collected in polythene bags to prevent the contamination and water evaporation. Collected soil samples were sieved. 10g of even sized soil samples were mixed with 100ml of distilled water and kept on an orbital shaker for 30 minutes. 1ml of aliquot was taken from the above sample and ten-fold soil dilution series was followed up to10 The 0.1ml of aliquot taken from the 10 soil dilute was inoculated into semi solid medium and also washed solution of roots, leaf parts and distilled wash root parts were inoculated in to test tubes containing Nitrogen free Bromothymole blue (NfB) semisolid media. All the tubes were inoculated at ambient temperature 30 C to 32 C for 48 hrs and the growth was observed by formation of subsurface white pellicles. The pellicles of each sample were streaked on NfB solid media and incubated at ambient temperature (28 C). Purified Azospirillum isolates were named as 100 series and stock cultures were prepared for further studies. Isolated Azospirillum spp were examined for identification of their Gram reaction, shape and motility. 0.2 ml of isolated bacterial broth were inoculated into 20 ml of NfB semisolid medium and allowed to incubate at 32 C for 10 days. The efficiency of Nitrogen fixation was determined after10 days using micro kjeldahl analysis.
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Keywords
Agriculture, Export Agriculture, Tea Industrials, Tea Technology, Soil Science, Biofertilizer
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