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  • Diversity of methanotrophic bacteria in tropical upland soils under different land uses.

Diversity of methanotrophic bacteria in tropical upland soils under different land uses.

Applied and environmental microbiology (2005-07-08)
Claudia Knief, Supika Vanitchung, Narumon W Harvey, Ralf Conrad, Peter F Dunfield, Amnat Chidthaisong
ABSTRACT

Three upland soils from Thailand, a natural forest, a 16-year-old reforested site, and an agricultural field, were studied with regard to methane uptake and the community composition of methanotrophic bacteria (MB). The methane uptake rates were similar to rates described previously for forest and farmland soils of the temperate zone. The rates were lower at the agricultural site than at the native forest and reforested sites. The sites also differed in the MB community composition, which was characterized by denaturing gradient gel electrophoresis (DGGE) of pmoA gene fragments (coding for a subunit of particulate methane monooxygenase) that were PCR amplified from total soil DNA extracts. Cluster analysis based on the DGGE banding patterns indicated that the MB communities at the forested and reforested sites were similar to each other but different from that at the farmland site. Sequence analysis of excised DGGE bands indicated that Methylobacter spp. and Methylocystis spp. were present. Sequences of the "forest soil cluster" or "upland soil cluster alpha," which is postulated to represent organisms involved in atmospheric methane consumption in diverse soils, were detected only in samples from the native forest and reforested sites. Additional sequences that may represent uncultivated groups of MB in the Gammaproteobacteria were also detected.

MATERIALS
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Sigma-Aldrich
REDAccuTaq® LA DNA Polymerase, High fidelity Taq with inert dye, 10X buffer included