Yuki Amano
Gregory F. Slater
Robert F. Dias
Teruki Iwatsuki
Kevin W. Mandernack
Christopher T. Mills
2010
<p><span>Microorganisms are ubiquitous in deep subsurface environments, but their role in the global carbon cycle is not well-understood. The natural abundance δ</span><sup>13</sup><span>C and Δ</span><sup>14</sup><span>C values of microbial membrane phospholipid fatty acids (PLFAs) were measured and used to assess the carbon sources of bacteria in sedimentary and granitic groundwaters sampled from three boreholes in the vicinity of the Tono Uranium Mine, Gifu, Japan. Sample storage experiments were performed and drill waters analyzed to characterize potential sources of microbial contamination. The most abundant PLFA structures in all waters sampled were 16:0, 16:1ω7</span><i>c</i><span>, </span><i>cy</i><span>17:0, and 18:1ω7</span><i>c</i><span>. A PLFA biomarker for type II methanotrophs, 18:1ω8</span><i>c</i><span>, comprised 3% and 18% of total PLFAs in anoxic sedimentary and granitic waters, respectively, sampled from the KNA-6 borehole. The presence of this biomarker was unexpected given that type II methanotrophs are considered obligate aerobes. However, a bacterium that grows aerobically with CH</span><sub>4</sub><span> as the sole energy source and which also produces 56% of its total PLFAs as 18:1ω8</span><i>c</i><span> was isolated from both waters, providing additional evidence for the presence of type II methanotrophs. The Δ</span><sup>14</sup><span>C values determined for type II methanotroph PLFAs in the sedimentary (−861‰) and granite (−867‰) waters were very similar to the Δ</span><sup>14</sup><span>C values of dissolved inorganic carbon (DIC) in each water (∼−850‰). This suggests that type II methanotrophs ultimately derive all their carbon from inorganic sources, whether directly from DIC and/or from CH</span><sub>4</sub><span> produced by the reduction of DIC. In contrast, δ</span><sup>13</sup><span>C values of type II PLFAs in the sedimentary (−93‰) and granite (−60‰) waters indicate that these organisms use different carbon assimilation schemes in each environment despite very similar </span><span class="math"><span id="MathJax-Element-2-Frame" class="MathJax_SVG" data-mathml="<math xmlns="http://www.w3.org/1998/Math/MathML"><mrow is="true"><msup is="true"><mrow is="true"><mi mathvariant="normal" is="true">&#x3B4;</mi></mrow><mrow is="true"><mn is="true">13</mn></mrow></msup><msub is="true"><mrow is="true"><mtext is="true">C</mtext></mrow><mrow is="true"><msub is="true"><mrow is="true"><mtext is="true">CH</mtext></mrow><mrow is="true"><mn is="true">4</mn></mrow></msub></mrow></msub></mrow></math>"><span class="MJX_Assistive_MathML">δ13CCH4</span></span></span><span> values (∼−95‰) for each water. The δ</span><sup>13</sup><span>C</span><sub>PLFA</sub><span> values (−28‰ to −45‰) of non-methanotrophic bacteria in the KNA-6 LTL water do not clearly distinguish between heterotrophic and autotrophic metabolisms, but Δ</span><sup>14</sup><span>C</span><sub>PLFA</sub><span>values indicate that >65% of total bacteria filtered from the KNA-6 LTL water are heterotrophs. Ancient Δ</span><sup>14</sup><span>C values (∼−1000‰) of some PLFAs suggest that many heterotrophs utilize ancient organic matter, perhaps from lignite seams within the sedimentary rocks. The more negative range of δ</span><sup>13</sup><span>C</span><sub>PLFA</sub><span> values determined for the KNA-6 granitic water (−42‰ to −66‰) are likely the result of a microbial ecosystem dominated by chemolithoautotrophy, perhaps fuelled by abiogenic H</span><sub>2</sub><span>. Results of sample storage experiments showed substantial shifts in microbial community composition and δ</span><sup>13</sup><span>C</span><sub>PLFA</sub><span>values (as much as 5‰) during 2–4 days of dark, refrigerated, aseptic storage. However, water samples collected and immediately filtered back in the lab from freshly drilled MSB-2 borehole appeared to maintain the same relative relationships between δ</span><sup>13</sup><span>C</span><sub>PLFA</sub><span> values for sedimentary and granitic host rocks as observed for samples directly filtered under artesian flow from the KNA-6 borehole of the Tono Uranium Mine.</span></p>
application/pdf
10.1016/j.gca.2010.03.016
en
Elsevier
Microbial carbon cycling in oligotrophic regional aquifers near the Tono Uranium Mine, Japan as inferred from δ13C and Δ14C values of in situ phospholipid fatty acids and carbon sources
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