Luo Y R, Tian Y, Huang X, Yan C L, Hong H S, Lin G H, Zheng T L
Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystems, School of Life Sciences, Xiamen University, Xiamen 361005, China; State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361005, China.
Key Laboratory of the Ministry of Education for Coastal and Wetland Ecosystems, School of Life Sciences, Xiamen University, Xiamen 361005, China; State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361005, China.
Mar Pollut Bull. 2009 Aug;58(8):1159-1163. doi: 10.1016/j.marpolbul.2009.03.024. Epub 2009 May 5.
A microbial consortium was obtained by enrichment culture of sea water samples collected from Botan oil port in Xiamen, China, using the persistent high concentration of a mixture of polycyclic aromatic hydrocarbons enrichment strategy. Denaturing gradient gel electrophoresis (DGGE) was used to investigate the bacterial composition and community dynamic changes based on PCR amplification of 16S rRNA genes during batch culture enrichment. Using the spray-plate method, three bacteria, designated as BL01, BL02 and BL03, which corresponded to the dominant bands in the DGGE profiles, were isolated from the consortium. Sequence analysis showed that BL01, BL02 and BL03 were phylogenetically close to Ochrobactrum sp., Stenotrophomonas maltophilia and Pseudomonas fluorescens, respectively. The degradation of benzo(a)pyrene (BaP), a model high-molecular-weight polycyclic aromatic hydrocarbon (HMW PAH) compound was investigated using individual isolates, a mixture of the three isolates, and the microbial consortium (BL) originally isolated from the oil port sea water. Results showed that the order of degradative ability was BL>the mixture of the three isolates>individual isolates. BL degraded 44.07% of the 10 ppm BaP after 14 days incubation, which showed the highest capability for HMW PAH compound degradation.Our results revealed that this high selective pressure strategy was feasible and effective in enriching the HMW PAH-degraders from the original sea water samples.
采用多环芳烃混合物持续高浓度富集策略，对采集自中国厦门博坦油港的海水样品进行富集培养，获得了一个微生物群落。在分批培养富集过程中，基于16S rRNA基因的PCR扩增，利用变性梯度凝胶电泳（DGGE）研究细菌组成和群落动态变化。采用涂布平板法，从该群落中分离出与DGGE图谱中优势条带相对应的三株细菌，分别命名为BL01、BL02和BL03。序列分析表明，BL01、BL02和BL03在系统发育上分别与慢生根瘤菌属、嗜麦芽窄食单胞菌和荧光假单胞菌相近。使用单个菌株、三株菌株的混合物以及最初从油港海水中分离出的微生物群落（BL），研究了典型的高分子量多环芳烃（HMW PAH）化合物苯并(a)芘（BaP）的降解情况。结果表明，降解能力顺序为BL>三株菌株的混合物>单个菌株。培养14天后，BL降解了10 ppm BaP的44.07%，显示出对HMW PAH化合物最高的降解能力。我们的结果表明，这种高选择性压力策略在从原始海水样品中富集HMW PAH降解菌方面是可行且有效的。
