Total N generally measured after wet digestion using the well known Kjeldahl method. This procedure involves three steps namely, digestion, distillation and titration. The  air dried (after removing root bits ) rhizosphere soil samples is digested in concentrated sulfuric acid with a catalyst mixture to raise the boiling temperature and to promote the conversion from organic-N to ammonium-N,. Ammonium-N from the digest is obtained by steam distillation, using strong alkali (40% NaOH) to raise the pH. The distillate is collected in saturated boric acid solution containing mixed indicator, and titrated with saturated acid.

Mineralizable N is estimated by Alkali Permanganate method by Subbiah and Asija (1956). Soil is distillated in the presence of excess of alkaline permanganate. The nascent oxygen liberated by KMNO4 in the presence of NaOH oxidizes readily oxidisable fraction of organic matter. During oxidation organic C is lost as CO2 and organic N is transformed in to NH3. The NH3 formed remains unstable in alkaline medium and it is readily distilled at boiling temperature in the distillation unit. The distillate containing NH3 is collected in 2% boric acid solution. The ammonia reacts with boric acid forming ammonium borate which is neutralized by titration with standard acid using mixed indicator.

NH4-N in soil extract you may estimate by Nessler's method: Ammonia react with potassium mercuric iodide (Nessler,s reagent) forming an orange coloured complex. The intensity of the colour is proportional to the concentration of NH4 present in it. The optical density of the coloured complex is measured at 420nm wave length using Spectrophotometer. The concentration of ammonia in soil extract is estimated by calibration curve prepared with standards.

Thanks very much sir for your kind attention and help

Dear Dr.Tarafdar,please see the key words"N fractions in rhizosphere soil'.Apart from routine total,mineralizable (chemical )  and mineral N in soil,are there any specific N fractions (present in labile or moderately labile carbon or specific compounds like aminoacids ,hexoseamine etc) in rhizosphere which can be extracted and estimated?.These fractions may contribute to N mineralization in rhizosphere and N availabilty to plant during crop growth.

Dr. Rao, N content in the rhizosphere are mostly mineral N, the method of which I have already explained. Mineral N (NH4+ and NO3-) can also be determined by using 2M KCl as the extracting solution in 1:5 (soil:KCl solution) ratio. Ammonium and nitrate plus nitrite are determined by steam distillation of ammonia, using MgO for NH4+ and Devarda's Alloy for NO3- (Bremner and keeney, 1965). The distillate is collected in 2% boric acid and titrate with dilute acid. This method determines dissolved and adsorbed forms of NH4+, NO3- and NO2- in the rhizosphere soils/soils. The sum of ammonium and nitrate determined by this method is refereed to as Mineral-N ( Keeney and Nelson, 1982, Buresh et al. 1982).

Sir, determination of ammoniacal and nitrate fractions of N using Bremner and Keeney (1965) method found to be more feasible. 

Thanks dear Dr Meera