Microbial community of a volcanic mudspring in the Philippines as revealed by 16S rDNA sequence analysis and fluorescence in situ hybridization

• Published in: World journal of microbiology & biotechnology Vol. 27; no. 4; pp. 859 - 867
• Main Authors: Lantican, Nacita B, Diaz, Ma. Genaleen Q, Cantera, J. Jason L, de los Reyes, Francis L. III, Raymundo, Asuncion K
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : Springer Netherlands 01-04-2011; Springer Netherlands; Springer; Springer Nature B.V
• Subjects: 16S rDNA; Applied Microbiology; Archaea; Bacteria; Biochemistry; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Cloning; Crenarchaeota; Environmental conditions; Environmental DNA; Environmental Engineering/Biotechnology; Euryarchaeota; Fluorescence; Fluorescence in situ hybridization; Fundamental and applied biological sciences. Psychology; Genetic testing; Hybridization; Laboratories; Leaching; Life Sciences; Microbiology; Microorganisms; Mudspring; Original Paper; Phylogenetics; Plasmids; Soil sciences; Studies; Sulfolobus tokodaii; Thermophiles; Volcanology; Environmental DNA; Mudspring; 16S rDNA; Thermophiles; Ribosomal DNA; Fluorescence in situ hybridization; 16S-RNA; Thermophily; Microbial community
• ISSN: 0959-3993; 1573-0972
• DOI: 10.1007/s11274-010-0528-y

Mt. Makiling Mudspring in Laguna, Philippines is a thermophilic, acidophilic environment that previously has been shown to harbor novel microorganisms. We assessed the microbial community that exists at this volcanic mudspring using 16S rRNA-based approaches. DNA was extracted from solfataric soils and sediments taken from Mudspring. The 16S rDNA was PCR amplified using universal (519F-1392R) and archaeal-specific (23FPL-1391R) primer pairs, cloned, and sequenced. Phylogenetic analysis of the cloned 16S rDNA showed that eleven clones clustered with, and therefore related to Sulfolobus tokodaii 7 and two clones clustered with S. solfataricu, S. shibatae and S. islandicus. Three clone sequences were related to those found in thermophilic chalcopyrite (CuFeS₂), a copper sulfuric ore from bioleaching reactors. One clone had low similarity (95% identity) with uncultured archaeon clone KOZ184. Fluorescence in situ hybridization (FISH) analysis revealed that about 71% of the microbial community present in the Mudspring belong to domain Archaea of which 63% were Crenarchaeota and 8% were Euryarchaeota. Seventeen percent (17%) of the population consisted of bacteria as indicated by the positive hybridization with the BACT338 probe, and the remaining 12% are unidentified. This study is the first attempt to use molecular techniques in any environment in the Philippines.