Application of Molecular Approach Towards Biological Nitrogen Fixation (BNF) in Azad Kashmir

Abstract

Increasing the agricultural production from marginal soils of Azad Jammu and Kashmir represent the real challenge for closing the wide gap between production and consumption of food. Most of the area of the state is hilly, the soils are shallow and devoid of vegetative cover. Due to indiscriminate deforestation and over grazing the soil erosion is destroying the soil, an important natural resource apart from other constituent of the environment, like soil micro-fauna and flora. Because of soil erosion and other environmental problem, the soil is subjected to low fertility, drought, low temperature, and low pH. The land holdings are small and the use of fertilizers is limited because of economical, atmospheric and environmental reservation. Biological nitrogen fixation (BNF) using biological agents like Rhizobium and pasture legumes in conjunction with pasture species of perennial grasses as a cover crop, holds the key to maximize the agricultural production, to ameliorate the soil conditions and to reduce the water erosion. Conventional uneconomic agricultural practices need diversification as the tillage practices on steep hills are not desirable. The establishment and the improvement of pasture lands using species of grasses and legumes that provide year-round green fodder with improved strains of Rhizobium for BNF look very much promising for the establishment of diary and sheep farming will help to establish the allied industries and the creation of job opportunities for the local population on suitable basis. Efforts to improve the species of grasses and legumes were already under way and the present project was initiated to collect, characterize, test and to manipulate the locally established strains of Rhizobium from various ecological conditions of the area. The Rhizobium isolates collected from various ecological conditions were characterized using convectional and molecular techniques. The conventional techniques like the antibiotic resistance and in vitro and in vivo nodulation tests proved that their exists diversity among the isolates, the isolates showed great potential for BNF under natural condition, however, their potential could only be exploited if their genetic and biochemical markers are identified. The molecular techniques like plasmid profiles the cellular protein profiles were investigated for identify some marker genes and their protein products for practical BNF and for the manipulation of DNA based markers for better activity of the bacteria in plant nodules. Comparisons based on plasmid and protein profiler indicated very interesting information. The isolates differed in nodulation test also showed variability I their plasmid and profile profiles. The relation between plasmid and protein profiles was interesting because its decrepit the presence of the variable gene on plasmids, which produce their influence in the form of protein products. These proteins may have their role against the environmental stresses in which the bacteria are adopted. The status of the gene once confirmed would be of great importance because plasmid is easy to handle, easy to manipulate and easy to transfer from one strain to another strain which may have otherwise desirability in BNF programme.