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We combined mercury geochemistry, hgcA gene cloning, rRNA methods, and metagenomics to compare microbial communities associated with MeHg production in two sulfate-impacted lakes on Minnesota\u2019s Mesabi Iron Range. The two lakes represent regional endmembers among sulfate-impacted sites in terms of their dissolved sulfide concentrations and MeHg production potential. rRNA amplicon sequencing indicates that sediments and anoxic bottom waters from both lakes contained diverse communities with multiple clades of sulfate reducing Deltaproteobacteria and Clostridia. In hgcA gene clone libraries, however, hgcA sequences were from taxa associated with methanogenesis and iron reduction in addition to sulfate reduction, and the most abundant clones were from unknown groups. We therefore applied metagenomics to identify the unknown populations in the lakes with the capability to methylate mercury, and reconstructed 27 genomic bins with hgcA. Some of the most abundant potential methylating populations were from phyla that are not typically associated with MeHg production, including a relative of the Aminicenantes (formerly candidate phylum OP8) and members of the Kiritimatiellaeota (PVC superphylum) and Spirochaetes that, together, were more than 50% of the potential methylators in some samples. These populations do not have genes for sulfate reduction, and likely degrade organic compounds by fermentation or other anaerobic processes. Our results indicate that previously unrecognized populations with hgcAB are abundant and may be important for MeHg production in some freshwater ecosystems.<\/jats:p>","DOI":"10.1038\/s41396-019-0376-1","type":"journal-article","created":{"date-parts":[[2019,2,26]],"date-time":"2019-02-26T12:05:47Z","timestamp":1551182747000},"page":"1659-1675","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":87,"title":["Molecular evidence for novel mercury methylating microorganisms in sulfate-impacted lakes"],"prefix":"10.1093","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4556-0418","authenticated-orcid":false,"given":"Daniel S","family":"Jones","sequence":"first","affiliation":[{"name":"BioTechnology Institute, University of Minnesota , St. Paul, MN, USA"},{"name":"Department of Earth Sciences, University of Minnesota , Minneapolis, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriel M","family":"Walker","sequence":"additional","affiliation":[{"name":"Department of Earth Sciences, University of Minnesota , Minneapolis, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathan W","family":"Johnson","sequence":"additional","affiliation":[{"name":"Department of Civil Engineering, University of Minnesota Duluth , Duluth, MN, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8538-5138","authenticated-orcid":false,"given":"Carl P J","family":"Mitchell","sequence":"additional","affiliation":[{"name":"Department of Physical and Environmental Sciences, University of Toronto \u2013 Scarborough , Toronto, ON, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jill K","family":"Coleman Wasik","sequence":"additional","affiliation":[{"name":"Department of Plant and Earth Science, University of Wisconsin River 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