Archaeal Community Revealed by 16s rRNA and Fluorescence in situ Hybridization in a Sulphuric Hydrothermal Hot Spring, Northern Taiwan |
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Authors: | Chang-Chai Ng Chen-Chin Chang Yuan-Tay Shyu |
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Institution: | (1) Institute of Horticulture, National Taiwan University, No.140, Keelung Road Section 4, Taipei, 10600, Taiwan ROC;(2) Department of Home Economics, Tainan Woman College of Arts and Technology, Tainan, Taiwan |
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Abstract: | Summary The archaeal community composition of Yangmingshan National Park in northern Taiwan was investigated by 16S rRNA and fluorescence
in situ hybridization (FISH). Optimization of tetrameric restriction enzyme (TRE) was performed to achieve efficient digestion and
differentiation in the restriction fragment length polymorphism (RFLP) fragments, and AciI, BstUI and RsaI were shown to be the optimal TREs for TRE-RFLP. Nine clones were obtained in the studies, with clones M70 and M6 being found
to be phylogenetically affiliated to Sulfolobus and Caldisphaera in domain Crenarchaeota, respectively, whereas seven other clones were found to be affiliated to an uncultured and unidentified
archaeon isolated from thermoacidic environments. In FISH, soil and water region cells were hybridized with DAPI (4′, 6-diamidino-2-phenylindole)
and specific fluorescently labelled probes. 15.69 and 7.16% of the DAPI-stained cells hybridized with universal archaeal probe
ARC915 and sulphate-reducing bacterial probe SRB385, respectively. |
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Keywords: | 16S rRNA archaea FISH microbial community thermoacidic hot spring |
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