Phylogenetic Analysis of Methanogenic Enrichment Cultures Obtained from Lonar Lake in India: Isolation of <Emphasis Type="Italic">Methanocalculus</Emphasis> sp. and <Emphasis Type="Italic">Methanoculleus</Emphasis> sp. |
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Authors: | Venkata Prasad Surakasi Aijaz Ahmad Wani Yogesh S Shouche Dilip R Ranade |
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Institution: | (1) Division of Microbial Sciences, Agharkar Research Institute, G. G. Agarkar Road, Pune, 411004, India;(2) Molecular Biology Unit, National Center for Cell Science, Ganeshkhind, University of Pune, Pune, 411007, India |
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Abstract: | The diversity of methanogenic archaea in enrichment cultures established from the sediments of Lonar Lake (India), a soda
lake having pH ≈ 10, was investigated using 16S rDNA molecular phylogenetic approach. Methanogenic enrichment cultures were
developed in a medium that simulated conditions of soda lake with three different substrates viz., H2:CO2, sodium acetate, and trimethylamine (TMA), at alkaline pH. Archaeal 16S rRNA clone libraries were generated from enrichment
cultures and 13 RFLP groups were obtained. Representative sequence analysis of each RFLP group indicated that the majority
of the 16S rRNA gene sequences were phylogenetically affiliated with uncultured Archaea. Some of the groups may belong to
new archaeal genera or families. Three RFLP groups were related to Methanoculleus sp, while two related to Methanocalculus sp. 16S rRNA gene sequences found in Lonar Lake were different from sequences reported from other soda lakes and more similar
to those of oil reservoirs, palm oil waste treatment digesters, and paddy fields. In culture-based studies, three isolates
were obtained. Two of these were related to Methanoculleus sp. IIE1 and one to Methanocalculus sp. 01F97C. These results clearly show that the Lonar Lake ecosystem harbors unexplored methanogens. |
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