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Authors: Yohda, Masafumi and Yagi, Osami and Takechi, Ayane and Kitajima, Mizuki and Matsuda, Hisashi and Miyamura, Naoaki and Aizawa, Tomoko and Nakajima, Mutsuyasu and Sunairi, Michio and Daiba, Akito and Miyajima, Takashi and Teruya, Morimi and Teruya, Kuniko and Shiroma, Akino and Shimoji, Makiko and Tamotsu, Hinako and Juan, Ayaka and Nakano, Kazuma and Aoyama, Misako and Terabayashi, Yasunobu and Satou, Kazuhito and Hirano, Takashi

A Dehalococcoides-containing bacterial consortium that performed dechlorination of 0.20 mM cis-1,2-dichloroethene to ethene in 14 days was obtained from the sediment mud of the lotus field. To obtain detailed information of the consortium, the metagenome was analyzed using the short-read next-generation sequencer SOLiD 3. Matching the obtained sequence tags with the reference genome sequences indicated that the Dehalococcoides sp. in the consortium was highly homologous to Dehalococcoides mccartyi CBDB1 and BAV1. Sequence comparison with the reference sequence constructed from 16S rRNA gene sequences in a public database showed the presence of Sedimentibacter, Sulfurospirillum, Clostridium, Desulfovibrio, Parabacteroides, Alistipes, Eubacterium, Peptostreptococcus and Proteocatella in addition to Dehalococcoides sp. After further enrichment, the members of the consortium were narrowed down to almost three species. Finally, the full-length circular genome sequence of the Dehalococcoides sp. in the consortium, D. mccartyi IBARAKI, was determined by analyzing the metagenome with the single-molecule DNA sequencer PacBio RS. The accuracy of the sequence was confirmed by matching it to the tag sequences obtained by SOLiD 3. The genome is 1,451,062 nt and the number of CDS is 1566, which includes 3 rRNA genes and 47 tRNA genes. There exist twenty-eight RDase genes that are accompanied by the genes for anchor proteins. The genome exhibits significant sequence identity with other Dehalococcoides spp. throughout the genome, but there exists significant difference in the distribution RDase genes. The combination of a short-read next-generation DNA sequencer and a long-read single-molecule DNA sequencer gives detailed information of a bacterial consortium. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Journal: Journal of bioscience and bioengineering
DOI: 10.1016/j.jbiosc.2014.12.001
Year: 2015

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