Phylogenetic analysis of methanogens in sheep rumen ecosystem and detection of Methanomicrobium mobile By fluorescence in situ hybridization.

Bioscience, biotechnology, and biochemistry ·Vol. 64 ·No. 8 ·2000-08-00 ·Pages 1737-42

Yanagita K, Kamagata Y, Kawaharasaki M, Suzuki T, Nakamura Y, Minato H

Abstract

The population of methanogens in the sheep rumen microbial ecosystem was studied by using 16S rDNA cloning analysis, epifluorescence microscopy (which detects autofluorescence of a specific cofactor F420 in methanogens) and the 16S rRNA-targeted in situ hybridization technique. The 16S rDNA clone libraries were constructed by PCR amplification with an Archaea-specific primer set and partial sequencing of the clonal 16S rDNAs was done. Phylogenetic analysis indicated that the clones were affiliated with Methanomicrobium ruminantium and mobile, Methanobrevibacter smithii. Epifluorescence microscopy (F420 autofluorescence) and in situ hybridization by using a newly designed M. mobile-specific 16S rRNA-targeted oligonucleotide probe found that methanogens accounted for approximately 3.6% of total ruminal microorganisms and approximately 54% of the total methanogens were M. mobile.

MeSH Terms

Animals DNA, Ribosomal/chemistry Ecosystem Euryarchaeota/classification,genetics Gene Library In Situ Hybridization, Fluorescence/veterinary Methanomicrobiaceae/isolation & purification Molecular Sequence Data Phylogeny RNA, Ribosomal, 16S/genetics Rumen/microbiology Sheep

Chemicals

DNA, Ribosomal RNA, Ribosomal, 16S

Authors & Affiliations

6 authors, click to expand affiliations / ORCID

Yanagita K

National Institute of Bioscience and Human Technology, Agency of Industrial Science and Technology, Tsukuba, Ibaraki, Japan.

Kamagata Y

Kawaharasaki M

Suzuki T

Nakamura Y

Minato H

Article Info

Journal

Bioscience, biotechnology, and biochemistry

Abbr.

Biosci Biotechnol Biochem

ISSN

0916-8451

DOI

10.1271/bbb.64.1737

Published

2000-08-00

Pages

1737-42

Language

English

Region

England

NLM ID

9205717

Subset

Databases

GENBANK

AB035622, AB035623, AB035624, AB035625, AB035626, AB035627, AB035628, AB035629, AB035630, AB035631, AB035632, AB035633, AB035634, AB035635, AB035636, AB035637, AB035638, AB035639, AB035640, AB035641

External Links

PubMed source DOI: 10.1271/bbb.64.1737 View journal details

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Phylogenetic analysis of methanogens in sheep rumen ecosystem and detection of Methanomicrobium mobile By fluorescence in situ hybridization.

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