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Modified sublimation to isolate phenanthrene-degrading bacteria of the genera Sphingomonas and Burkholderia from Xiamen oil port
Authors:X. Huang   Y. Tian   Y.R. Luo   H.J. Liu   Wei Zheng  T.L. Zheng  
Affiliation:aCollege of Oceanography and Environmental Science, State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361005, China;bSchool of Life Sciences, Key Lab for Subtropical Wetland Ecosystem Research, Ministry of Education, Xiamen University, Xiamen 361005, China
Abstract:Sublimation was developed by Alley and Brown (2000) in order to isolate bacterial strains that were capable of degrading water insoluble compounds. In this study, sublimation was modified by the use of nutritional agar plates, instead of mineral salt agar, to isolate phenanthrene-degrading bacteria from a mixed culture that had been enriched under the selective pressure of high phenanthrene content. Five strains were obtained with different morphology and degradation ability. Based on the 16S rDNA sequence, two of them were classified as species of the genus Sphingomonas; the others as species of the genus Burkholderia. Denaturing gradient gel electrophoresis (DGGE) was introduced to detect dynamic changes in the bacterial community during enrichment batch culture, and to determine any correlation between the five isolates and the phenanthrene-degrading consortium. The DGGE profile indicated that these five isolates corresponded to four dominant bands of the consortium. Compared to traditional means of isolation, we concluded that modified sublimation is effective and more convenient.
Keywords:Sublimation   Denaturing gradient gel electrophoresis   Sphingomonas   Burkholderia   Phenanthrene
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