Determining the Reliability of Measuring Mercury Cycling Gene Abundance with Correlations with Mercury and Methylmercury Concentrations

Christensen, Geoffrey A.; Gionfriddo, Caitlin M.; King, Andrew J.; Moberly, James G.; Miller, Carrie; Somenahally, Anil C.; Callister, Stephen J.; Brewer, Heather M.; Podar, Mircea; Brown, Steven D.; Palumbo, Anthony V.; Brandt, Craig C.; Wymore, Ann M.; Brooks, Scott C.; Hwang, C; Fields, Matthew W.; Wall, Judy D.; Gilmour, Cynthia C.; Elias, Dwayne A.

doi:10.1021/acs.est.8b06389

Title: Determining the Reliability of Measuring Mercury Cycling Gene Abundance with Correlations with Mercury and Methylmercury Concentrations

Journal Article · Tue Aug 06 00:00:00 EDT 2019 · Environmental Science & Technology

DOI:https://doi.org/10.1021/acs.est.8b06389· OSTI ID:1572494

Christensen, Geoffrey A. ^[1]; Gionfriddo, Caitlin M. ^[1]; King, Andrew J. ^[1]; Moberly, James G. ^[1]; Miller, Carrie ^[2]; Somenahally, Anil C. ^[3]; Callister, Stephen J. ^[4];

^[4]; Podar, Mircea ^[5]; Brown, Steven D. ^[1]; Palumbo, Anthony V. ^[1]; Brandt, Craig C. ^[1]; Wymore, Ann M. ^[1]; Brooks, Scott C. ^[1]; Hwang, C ^[6]; Fields, Matthew W. ^[6]; Wall, Judy D. ^[7]; Gilmour, Cynthia C. ^[8]; Elias, Dwayne A. ^[9]

Oak Ridge National Laboratory
Troy State University
Texas A & M University
BATTELLE (PACIFIC NW LAB)
ORNL
Montana State University
University of Missouri, Columbia
Smithsonian Enviromental Research Center
OAK RIDGE NATIONAL LAB

Methylmercury (MeHg) is a bioaccumulative toxic contaminant in many ecosystems but factors governing its production are poorly understood. Recent work has shown that the anaerobic microbial conversion of mercury (Hg) to MeHg requires the Hg-methylation genes hgcAB and that these genes can be used as biomarkers in PCR-based estimators of Hg-methylator abundance. In an effort to determine reliable methods for assessing hgcA abundance and diversity and linking it to MeHg concentrations, multiple approaches were compared including metagenomic shotgun sequencing, 16S rRNA gene pyrosequencing and cloning/sequencing hgcAB gene products. Hg-methylator abundance was also determined by quantitative hgcA qPCR amplification and metaproteomics for comparison to the above measurements. Samples from eight Hg-contaminated sites were examined over a range of total Hg (HgT; 0.03–14 mg kg-1 wet wt. soil) and MeHg (0.05–27 µg kg-1 wet wt. soil) concentrations. In the metagenome and amplicon sequencing of hgcAB diversity, the Deltaproteobacteria were the dominant Hg-methylators, while Firmicutes and methanogenic Archaea were typically ~50% less abundant. This was consistent with metaproteomics estimates where the Deltaproteobacteria were consistently higher. The 16S rRNA gene pyrosequencing did not have sufficient resolution to identify hgcAB+ species. Metagenomic and hgcAB results were similar for Hg-methylator diversity and showed a tractable relationship between Hg-methylating bacteria and soil Hg concentrations among similar environmental samples. However, clade-specific qPCR-based approaches for hgcA are only appropriate when comparing the abundance of a particular clade across various samples.

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Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

DOE Contract Number:: AC05-76RL01830

OSTI ID:: 1572494

Report Number(s):: PNNL-SA-145153

Journal Information:: Environmental Science & Technology, Vol. 53, Issue 15

Country of Publication:: United States

Language:: English

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An Improved hgcAB Primer Set and Direct High-Throughput Sequencing Expand Hg-Methylator Diversity in Nature Gionfriddo, Caitlin M.; Wymore, Ann M.; Jones, Daniel S. Frontiers in Microbiology, Vol. 11 https://doi.org/10.3389/fmicb.2020.541554	journal	October 2020

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hgcAB
16s sequencinv

Title: Determining the Reliability of Measuring Mercury Cycling Gene Abundance with Correlations with Mercury and Methylmercury Concentrations

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