Huang X, Tian Y, Luo Y R, Liu H J, Zheng Wei, Zheng T L
College of Oceanography and Environmental Science, State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361005, China.
Mar Pollut Bull. 2008;57(6-12):538-43. doi: 10.1016/j.marpolbul.2007.12.013. Epub 2008 May 27.
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.
升华法由艾利和布朗于2000年开发,用于分离能够降解水不溶性化合物的细菌菌株。在本研究中,对升华法进行了改进,使用营养琼脂平板而非无机盐琼脂,从在高菲含量的选择压力下富集的混合培养物中分离菲降解细菌。获得了五株形态和降解能力不同的菌株。基于16S rDNA序列,其中两株被归类为鞘氨醇单胞菌属的物种;其他的为伯克霍尔德菌属的物种。引入变性梯度凝胶电泳(DGGE)来检测富集分批培养过程中细菌群落的动态变化,并确定这五株分离菌与菲降解菌群之间的任何相关性。DGGE图谱表明这五株分离菌对应于该菌群的四个优势条带。与传统的分离方法相比,我们得出结论,改进后的升华法有效且更方便。