title: Bio-Augmentation of Cupriavidus sp. CY-1 into 2,4-D Contaminated Soil: Microbial Community Analysis by Culture Dependent and Independent Techniques.
abstract: 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.
authors.affiliation: Course of Chemical and Biological Engineering, Division of Sustainable and Environmental Engineering, College of Environmental Technology, Muroran Institute of Technology, 27-1 Mizumoto, Muroran, 050-8585, Japan.
authors.forename: Young-Cheol
authors.initials: YC
authors.lastname: Chang
authors.affiliation: Course of Chemical and Biological Engineering, Division of Sustainable and Environmental Engineering, College of Environmental Technology, Muroran Institute of Technology, 27-1 Mizumoto, Muroran, 050-8585, Japan.
authors.forename: M Venkateswar
authors.initials: MV
authors.lastname: Reddy
authors.affiliation: Course of Chemical and Biological Engineering, Division of Sustainable and Environmental Engineering, College of Environmental Technology, Muroran Institute of Technology, 27-1 Mizumoto, Muroran, 050-8585, Japan.
authors.forename: Honoka
authors.initials: H
authors.lastname: Umemoto
authors.affiliation: Course of Chemical and Biological Engineering, Division of Sustainable and Environmental Engineering, College of Environmental Technology, Muroran Institute of Technology, 27-1 Mizumoto, Muroran, 050-8585, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Sato
authors.affiliation: Seoul Metropolitan Government Research Institute of Public Health and Environment, 202-3 Yangjae-dong, Seocho-gu, Seoul, 137-893, Republic of Korea.
authors.forename: Mi-Hye
authors.initials: MH
authors.lastname: Kang
authors.affiliation: Graduate School of Medicine, Kyoto University, Yoshidakonoe-cho, Sakyo-ku, Kyoto-shi, Kyoto, 606-8501, Japan.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Yajima
authors.affiliation: Course of Chemical and Biological Engineering, Division of Sustainable and Environmental Engineering, College of Environmental Technology, Muroran Institute of Technology, 27-1 Mizumoto, Muroran, 050-8585, Japan.
authors.forename: Shintaro
authors.initials: S
authors.lastname: Kikuchi
mesh_terms: D015084:2,4-Dichlorophenoxyacetic Acid / Q000378:metabolism*; D001673:Biodegradation, Environmental; D044822:Biodiversity; D002851:Chromatography, High Pressure Liquid; D049921:Cupriavidus / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics; D058645:Denaturing Gradient Gel Electrophoresis; D017753:Ecosystem; D008401:Gas Chromatography-Mass Spectrometry; D006540:Herbicides / Q000378:metabolism*; D009566:Nitrates / Q000737:chemistry; D012150:Polymorphism, Restriction Fragment Length; D017680:Potassium Compounds / Q000737:chemistry; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012965:Sodium Chloride / Q000737:chemistry; D012987:Soil / Q000737:chemistry; D012988:Soil Microbiology; D012989:Soil Pollutants / Q000378:metabolism*
chemical_list: D004269:DNA, Bacterial; D006540:Herbicides; D009566:Nitrates; D017680:Potassium Compounds; D012336:RNA, Ribosomal, 16S; D012987:Soil; D012989:Soil Pollutants; D015084:2,4-Dichlorophenoxyacetic Acid; D012965:Sodium Chloride; C023844:potassium nitrate