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Cultivation and characterization of the bacterial assemblage of epsomic Basque Lake, BC

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Abstract

Athalassohaline waters that are rich in divalent ions are good analogues for the chemical environments of Mars and the ocean worlds. Sulfate salts, along with chlorides, are important in Mars regolith with Ca, Fe, Mg, and Na counterions. Certain lakes in the Pacific Northwest are saturated with MgSO4 as epsomite. Here we report on the microbial community of Basque Lake, BC, a group of playas that is saturated with MgSO4. More than 60 bacterial isolates were obtained from Basque Lake soils by enrichment culture and repetitive streak-plating using media containing 10% (~ 1.7 M) NaCl or 50% (~ 2 M) MgSO4. Most of the isolates (~ 75%) were Gram-positive, motile, and produced endospores. Isolates related to Marinococcus halophilus and Virgibacillus marismortui dominated the collection. Halomonas and Salinivibrio were Gram-negative genera found at Basque Lake. Nearly all of the Basque Lake isolates grew at 50% MgSO4, with 65% growing at 60% MgSO4. Several isolates could grow in saturated (67%) MgSO4 (aw = 0.90). All of the isolates grew at 10% NaCl with 70% growing at 20% salinity (~ 3.5 M NaCl; aw = 0.82). Basque Lake isolates grew better at basic pH than acidic pH, with 80% growing at pH 9 and 30% growing at pH 10. Only 20% of the isolates grew at pH 5. Numerical taxonomy dendrograms based on 44 phenetic characteristics showed a strong correspondence to phylogenetic trees constructed from 16S rRNA gene sequences. Pyrosequencing of 16S rRNA gene sequences from direct DNA extracts of Basque Lake soils recovered predominantly Proteobacteria (60%), Firmicutes (11%), and unclassified bacteria (27%). Microbes capable of growth under the extreme chemical conditions of Mars are a particular concern for forward planetary protection should they contaminate a spacecraft.

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Acknowledgements

The authors are thankful for the preliminary and supportive work done by Amer Al Soudi, Bishal Bista, John Dille, Timothy Eberl, Casper Fredsgaard, Brian Kilmer, Tony Mai, Donald Moore, Trista Newville, Kyle Rowe, Namrata Shrestha, and Karen Woltersdorf. We are grateful to Bruce Madu and Jim Britton (British Columbia Ministry of Energy and Mines; BCMEM) for collecting and documenting Basque Lake samples. We thank Fadi Aramouni (Kansas State University) for performing water activity measurements and Stephen Lindemann (Purdue University) for deep sequencing analyses. Preliminary accounts of this work have been presented previously (Crisler et al. 2010a, b, 2018; Kilmer et al. 2012). This work was supported by awards from National Aeronautics and Space Administration (NASA), Research Opportunities in Space and Earth Science (ROSES), Planetary Protection Research (09-PPR09-0004 and 14-PPR14-2-0002). Additional student support was from Kansas Institutional Development Award (IDeA) Networks of Biomedical Research Excellence (KINBRE), National Institute of General Medical Sciences (NIGMS), National Institutes of Health (NIH) (P20 GM103418). The content is solely the responsibility of the authors and does not necessarily represent the official views of BCMEM, KINBRE, NASA, NIGMS or NIH.

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  1. Department of Biological Sciences, Wichita State University, 26, 1845 Fairmount, Wichita, KS, 67260, USA

    James D. Crisler & Mark A. Schneegurt

  2. Planetary Protection Group, Jet Propulsion Laboratory, NASA, Pasadena, CA, USA

    Fei Chen

  3. Space Science Institute, Boulder, CO, USA

    Benton C. Clark

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JDC, FC, BCC and MAS conceived and designed the study. JDC, FC and MAS performed research and analyzed data. JDC and MAS wrote the paper.

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Correspondence to Mark A. Schneegurt.

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10482_2019_1244_MOESM1_ESM.png

Figure S1. Phylogenetic tree for Gram-positive bacteria from Basque Lake based on 16S rRNA gene sequences. GenBank accession numbers, sampling site, enrichment temperature, and enrichment medium (10% NaCl or 50% MgSO4) are given for each isolate. (PNG 40 kb)

10482_2019_1244_MOESM2_ESM.png

Figure S2. Phylogenetic tree for Gram-negative bacteria from Basque Lake based on 16S rRNA gene sequences. GenBank accession numbers, sampling site, enrichment temperature, and enrichment medium (10% NaCl or 50% MgSO4) are given for each isolate. (PNG 16 kb)

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Crisler, J.D., Chen, F., Clark, B.C. et al. Cultivation and characterization of the bacterial assemblage of epsomic Basque Lake, BC. Antonie van Leeuwenhoek 112, 1105–1119 (2019). https://doi.org/10.1007/s10482-019-01244-0

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