Urakawa H, Yoshida T, Nishimura M, Ohwada K
Ocean Research Institute, University of Tokyo, Japan.
Environ Microbiol. 2000 Oct;2(5):542-54. doi: 10.1046/j.1462-2920.2000.00137.x.
Depth-related changes in whole-community structure were evaluated in a coastal marine sediment using a molecular fingerprinting method, terminal restriction fragment length polymorphism (T-RFLP) analysis, and a chemotaxonomic technique (quinone profiling). Dendrograms derived from both T-RFLP analysis and quinone profiling indicated a significant variation in microbial community structure between the 0-2 cm layer and deeper layers. This corresponded to the dramatic change in the redox potential, acid-volatile sulphide-sulphur and bacterial numbers observed at 0-2 cm and 2-4 cm depths. A significant change in the number of terminal restriction fragments (T-RFs) was also detected at this transition depth. However, the change in major T-RFs with depth was not seen in electropherograms. The population changes were primarily variations in minor ribotypes. Most quinone homologues were detected at all depths, although the quinone composition changed with depth. Therefore, quinone profiling also suggested that the depth-related variation was primarily attributable to minor bacterial groups rather than change in the major population structure. 16S rDNA clone library analysis revealed that clones belonging to the genera Vibrio and Serratia predominated as major bacterial groups at all depths. Our data suggested that the sediment community might result from sedimentation effects of sinking particles. Overall, our results demonstrated that the combined methods of T-RFLP analysis and quinone profiling were effective for assessing depth-related microbial populations.
使用分子指纹识别方法、末端限制性片段长度多态性(T-RFLP)分析和化学分类技术(醌谱分析),对沿海海洋沉积物中与深度相关的整个群落结构变化进行了评估。从T-RFLP分析和醌谱分析得出的树形图表明,0-2厘米层和更深层之间的微生物群落结构存在显著差异。这与在0-2厘米和2-4厘米深度处观察到的氧化还原电位、酸挥发性硫化物-硫和细菌数量的剧烈变化相对应。在这个过渡深度也检测到末端限制性片段(T-RFs)数量的显著变化。然而,在电泳图中未观察到主要T-RFs随深度的变化。种群变化主要是次要核糖体类型的变化。尽管醌的组成随深度变化,但在所有深度都检测到了大多数醌同系物。因此,醌谱分析也表明,与深度相关的变化主要归因于次要细菌群体,而不是主要种群结构的变化。16S rDNA克隆文库分析表明,属于弧菌属和沙雷氏菌属的克隆在所有深度作为主要细菌群体占主导地位。我们的数据表明,沉积物群落可能是下沉颗粒沉降效应的结果。总体而言,我们的结果表明,T-RFLP分析和醌谱分析相结合的方法对于评估与深度相关的微生物种群是有效的。