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KMID : 0364820120480010042
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Korean Journal of Microbiology
2012 Volume.48 No. 1 p.42 ~ p.47
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Identification of Anaerobic Thermophilic Thermococcus Dominant in Enrichment Cultures from a Hydrothermal Vent Sediment of Tofua Arc
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Cha In-Tae
Kim So-Jeong
Kim Jong-Geol
Park Soo-Je
Jung Man-Young
Ju Se-Jong
Kwon Kae-Kyoung
Rhee Sung-Keun
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Abstract
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Hydrothermal vents (HTV) provide special environments for evolution of lives independent on solar energy. HTV
samples were gained from Tofua arc trench in Tonga, South Pacific. We investigated archaeal diversity enriched
using combinations of various electron donors (yeast extract and H2) and electron acceptors [Iron (III), elemental
sulfur (S0) and nitrate. PCR amplification was performed to detect archaeal 16S rRNA genes after the cultures were
incubated 65℃ and 80℃ for 2 weeks. The cultures showing archaeal growth were transferred using the
dilution-to-extinction method. 16S rRNA gene PCR-Denaturing Gradient Gel Electrophoresis was used to identify
the enriched archaea in the highest dilutions where archaeal growth was observed. Most of cultured archaea
belonged to genus of Thermococcus (T. alcaliphilius, T. litoralis, T. celer, T. barossii, T. thoreducens, T. coalescens)
with 98?99% 16S rRNA gene similarities. Interestingly, archaeal growth was observed in the cultures with Iron (III)
and nitrate as an electron acceptor. It was supposed that archaea might use the elemental sulfur generated from
oxidation of the reducing agent, sulfide. To cultivate diverse archaea excluding Thermococcus, it would be required
to use other reducing agents instead of sulfide
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KEYWORD
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Thermococcus, anaerobic archaea, enrichment culture, extinction dilution culture, hydrothermal vent
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