Affiliation:
1. Graduate School of Life Sciences, Tohoku University, Katahira, Aoba-ku, Sendai, Japan
Abstract
ABSTRACT
Anaerobic nitrogen-fixing consortia consisting of N
2
-fixing clostridia and diverse nondiazotrophic bacteria were previously isolated from various gramineous plants (K. Minamisawa, K. Nishioka, T. Miyaki, B. Ye, T. Miyamoto, M. You, A. Saito, M. Saito, W. Barraquio, N. Teaumroong, T. Sein, and T. Tadashi, Appl. Environ. Microbiol.
70:
3096-3102, 2004). For this work, clostridial populations and their phylogenetic structures in a stand of the grass
Miscanthus sinensis
in Japan were assessed by a 16S rRNA gene-targeted terminal restriction fragment length polymorphism (TRFLP) analysis combined with most-probable-number (MPN) counts. PCR primers and restriction enzymes were optimized for analyses of the plant clostridia. Clostridia were detected in strongly surface-sterilized leaves, stems, and roots of the plants at approximately 10
4
to 10
5
cells/g of fresh weight; they made up a large proportion of N
2
-fixing bacterial populations, as determined by MPN counts associated with an acetylene reduction assay. Phylogenetic grouping by MPN-TRFLP analysis revealed that the clostridial populations belonged to group II of cluster XIVa and groups IV and V of cluster I; this result was supported by a culture-independent TRFLP analysis using direct DNA extraction from plants. When phylogenetic populations from
M. sinensis
and the soil around the plants were compared, group II clostridia were found to exist exclusively in
M. sinensis
.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
85 articles.
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