Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
AMB Express. 2012 Sep 18;2(1):50. doi: 10.1186/2191-0855-2-50.
We recently established a method for isolating functional single cells from environmental samples using a micromanipulator (Functional single-cell (FSC) isolation), and applied it to the study of denitrifying bacteria in rice paddy soil (Ashida et al. 2010. Appl Microbiol Biotechnol 85:1211-1217). To further examine the advantages and possible disadvantages of the FSC method, we isolated denitrifying bacteria from the same rice paddy soil sample using both FSC and standard agar plate dilution (APD) methods and compared in this study. The proportion of denitrifying bacteria in the total isolates was more than 6-fold larger with FSC isolation (57.1%) compared with the APD method (9.2%). Denitrifying bacteria belonging to Alphaproteobacteria and Bacilli were commonly isolated using both methods, whereas those belonging to Betaproteobacteria, which had been found to be active in the denitrification-inductive paddy soil, were isolated only with the FSC method. On the other hand, Actinobacteria were only isolated using the APD method. The mean potential denitrification activity of the FSC isolates was higher than that of the APD isolates. Overall, FSC isolation was confirmed to be an excellent method for studying denitrifying bacteria compared with the standard agar plate dilution method.
我们最近建立了一种使用微操作器从环境样本中分离功能单细 胞的方法（功能单细胞（FSC）分离），并将其应用于稻田土壤中脱 氮细菌的研究（Ashida 等人，2010. Appl Microbiol Biotechnol 85:1211-1217）。为了进一步检查 FSC 方法的优点和可能的缺点，我们 使用 FSC 和标准琼脂平板稀释（APD）方法从同一稻田土壤样本中 分离脱氮细菌，并在本研究中进行了比较。与 APD 方法（9.2%）相 比，FSC 分离法获得的总分离物中脱氮细菌的比例高 6 倍以上（57.1%）。两种方法都能分离到属于 Alphaproteobacteria 和 Bacilli 的脱氮细 菌，而属于 Betaproteobacteria 的脱氮细菌在诱导稻田中具有活性，但只 能用 FSC 方法分离。另一方面，放线菌只能用 APD 方法分离。FSC 分离物的平均潜在脱氮活性高于 APD 分离物。总的来说，与标准 琼脂平板稀释法相比，FSC 分离法被证实是研究脱氮细菌的一种优 秀方法。
