RRC ID 5861
著者 Ashida N, Ishii S, Hayano S, Tago K, Tsuji T, Yoshimura Y, Otsuka S, Senoo K.
タイトル Isolation of functional single cells from environments using a micromanipulator: application to study denitrifying bacteria.
ジャーナル Appl Microbiol Biotechnol
Abstract We developed a novel method to isolate functionally active single cells from environmental samples and named it the functional single-cell (FSC) isolation method. This method is based on a combination of substrate-responsive direct viable counts, live-cell staining with 5-carboxyfluorescein diacetate acetoxymethyl ester, and micromanipulation followed by cultivation in a medium. To evaluate this method, we applied it to study a denitrifying community in rice paddy soil. Similar denitrifier counts were obtained by the conventional most probable number analysis and our FSC isolation method. Using the FSC isolation method, 37 denitrifying bacteria were isolated, some of which harbored copper-containing nitrite reductase gene (nirK). The 16S rRNA gene analysis showed that members belonging to the genera Azospirillum and Ochrobactrum may be the major denitrifiers in the rice paddy soil. These results indicate that the FSC isolation method is a useful tool to obtain functionally active single cells from environmental samples.
巻・号 85(4)
ページ 1211-7
公開日 2010-1-1
DOI 10.1007/s00253-009-2330-z
PMID 19936739
MeSH Azospirillum / classification Azospirillum / isolation & purification Bacteria / classification Bacteria / genetics Bacteria / isolation & purification* Bacteriological Techniques* DNA, Bacterial / genetics DNA, Ribosomal / genetics Fluoresceins Nitrates / metabolism Ochrobactrum / classification Ochrobactrum / isolation & purification Oryza* Phylogeny RNA, Ribosomal, 16S / genetics Soil Microbiology*
IF 3.53
引用数 52
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
リソース情報
一般微生物 JCM 20522 JCM 1224 JCM 2505 JCM 21446 JCM 21315 JCM 12830 JCM 21032 JCM 20264 JCM 5963 JCM 5965