This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Miyamoto, T. Articles by Minamisawa, K. Search for Related Content PubMed PubMed Citation Articles by Miyamoto, T. Articles by Minamisawa, K. Agricola Articles by Miyamoto, T. Articles by Minamisawa, K.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, November 2004, p. 6580-6586, Vol. 70, No. 11
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.11.6580-6586.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Novel Endophytic Nitrogen-Fixing Clostridia from the Grass Miscanthus sinensis as Revealed by Terminal Restriction Fragment Length Polymorphism Analysis

Graduate School of Life Sciences, Tohoku University, Katahira, Aoba-ku, Sendai, Japan

Received 25 February 2004/ Accepted 18 July 2004

Anaerobic nitrogen-fixing consortia consisting of N₂-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 10⁴ to 10⁵ cells/g of fresh weight; they made up a large proportion of N₂-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.

This article has been cited by other articles:

• Julien, M.-C., Dion, P., Lafreniere, C., Antoun, H., Drouin, P. (2008). Sources of Clostridia in Raw Milk on Farms. Appl. Environ. Microbiol. 74: 6348-6357 [Abstract] [Full Text]  
• Torok, V. A., Ophel-Keller, K., Loo, M., Hughes, R. J. (2008). Application of Methods for Identifying Broiler Chicken Gut Bacterial Species Linked with Increased Energy Metabolism. Appl. Environ. Microbiol. 74: 783-791 [Abstract] [Full Text]  
• Okazaki, S., Sugawara, M., Yuhashi, K.-I., Minamisawa, K. (2007). Rhizobitoxine-induced Chlorosis Occurs in Coincidence with Methionine Deficiency in Soybeans. ANN BOT (LOND) 100: 55-59 [Abstract] [Full Text]  
• Thies, J. E. (2007). Soil Microbial Community Analysis using Terminal Restriction Fragment Length Polymorphisms. Soil Sci. 71: 579-591 [Abstract] [Full Text]  
• Sameshima-Saito, R., Chiba, K., Hirayama, J., Itakura, M., Mitsui, H., Eda, S., Minamisawa, K. (2006). Symbiotic Bradyrhizobium japonicum Reduces N2O Surrounding the Soybean Root System via Nitrous Oxide Reductase. Appl. Environ. Microbiol. 72: 2526-2532 [Abstract] [Full Text]  
• You, M., Nishiguchi, T., Saito, A., Isawa, T., Mitsui, H., Minamisawa, K. (2005). Expression of the nifH Gene of a Herbaspirillum Endophyte in Wild Rice Species: Daily Rhythm during the Light-Dark Cycle. Appl. Environ. Microbiol. 71: 8183-8190 [Abstract] [Full Text]