Shigematsu Toru, Hayashi Mayumi, Kikuchi Isamu, Ueno Shigeaki, Masaki Haruhiko, Fujii Tomoyuki
Department of Food Science, Faculty of Applied Life Sciences, Niigata University of Pharmacy and Applied Life Sciences, Niigata, Japan.
FEMS Microbiol Lett. 2009 Apr;293(2):240-7. doi: 10.1111/j.1574-6968.2009.01536.x. Epub 2009 Mar 2.
A method for analyzing culture-dependent bacterial community structure by liquid cultivation was established using 96-well microplates. Using 96-well microplates, this method can easily provide accurate enumeration of viable microorganisms and simultaneous separation of bacteria, which allowed us to analyze the bacterial community. Bacteria in diluted surface seawater were separated using 96-well microplates and cultivated with 1/5 ZoBell 2216E liquid medium. The 98 cultures obtained were subsequently applied to phylogenetic analysis based on 16S rRNA gene sequences. The bacterial diversity, evaluated by the Shannon-Weaver index, was relatively small but comparable to previously reported bacterial communities of several environments. The most abundant group was the family Rhodobacteraceae, which has been frequently detected in marine environments. Most bacteria were phylogenetically related to bacteria or uncultured clones detected in marine environments, but distant from published species. The analysis of bacterial community structure by liquid cultivation would be useful as an alternative culture-dependent approach.
利用96孔微孔板建立了一种通过液体培养分析依赖培养的细菌群落结构的方法。使用96孔微孔板，该方法可以轻松地对活微生物进行准确计数并同时分离细菌，这使我们能够分析细菌群落。稀释后的表层海水中的细菌使用96孔微孔板进行分离，并用1/5的ZoBell 2216E液体培养基培养。随后将获得的98个培养物应用于基于16S rRNA基因序列的系统发育分析。通过香农-韦弗指数评估的细菌多样性相对较小，但与先前报道的几种环境中的细菌群落相当。最丰富的类群是红杆菌科，该科在海洋环境中经常被检测到。大多数细菌在系统发育上与在海洋环境中检测到的细菌或未培养克隆相关，但与已发表的物种距离较远。通过液体培养分析细菌群落结构作为一种替代的依赖培养的方法将是有用的。
