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In the present study, a 2,4-dichlorophenoxyacetic acid (2,4-D) degrading bacterial strain CY-1 was isolated from forest soil.Based on physiological, biochemical and 16S rRNA gene sequence analysis it was identified as Cupriavidus sp.CY-1.Further 2,4-D degradation experiments at different concentrations (200 to 800 mg/l) were carried out using CY-1.On the other hand, two different types of soils collected from two different sources were used for 2,4-D degradation studies.The isolated strain CY-1 was bio-augmented into 2,4-D contaminated soils to analyze its degradation ability.Culture independent methods like denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP), and culture dependent methods like colony forming units (CFU) and most probable number (MPN) were used to analyze the survivability of strain CY-1 in contaminated soil.Results of T-RFLP were coincident with the DGGE analysis.From the DGGE, T-RFLP, MPN and HPLC results it was concluded that strain CY-1 effectively degraded 2,4-D without disturbing the ecosystem of soil indigenous microorganisms.