Journal Article Bio-augmentation of cupriavidus sp. CY-1 into 2,4-D contaminated soil: Microbial community analysis by culture dependent and independent techniques

Chang, Young Cheol  ,  Reddy, M. Venkateswar  ,  Umemoto, Honoka  ,  Sato, Yuki  ,  Kang, Mi Hye  ,  Yajima, Yuka  ,  Kikuchi, Shintaro

10 ( 12 ) 2015-12-28 , Public Library of Science
In the present study, a 2, 4-dichlorophenoxyacetic acid (2, 4-D) degrading bacterial strain CY-1 was isolated from the 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-1) were carried out using CY-1. Effect of NaCl and KNO3 on 2, 4-D degradation was also evaluated. Degradation of 2, 4-D and the metabolites produced during degradation process were analyzed using high pressure liquid chromatography (HPLC) and GC-MS respectively. The amount of chloride ions produced during the 2, 4-D degradation were analyzed by Ion chromatography (IC) and it is stoichiometric with 2, 4-D dechlorination. Furthermore 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.

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