TY - JOUR T1 - Variations in microbial carbon sources and cycling in the deep continental subsurface JF - Geochimica et Cosmochimica Acta Y1 - 2016 A1 - Danielle N. Simkus A1 - Greg F. Slater A1 - Barbara Sherwood Lollar A1 - Kenna Wilkie A1 - Thomas L. Kieft A1 - Cara Magnabosco A1 - Maggie C.Y. Lau A1 - Michael J. Pullin A1 - Sarah B. Hendrickson A1 - K. Eric Wommack A1 - Eric G. Sakowski A1 - Esta van Heerden A1 - Olukayode Kuloyo A1 - Borja Linage A1 - Gaetan Borgonie A1 - Tullis C. Onstott AB - Abstract Deep continental subsurface fracture water systems, ranging from 1.1 to 3.3 km below land surface (kmbls), were investigated to characterize the indigenous microorganisms and elucidate microbial carbon sources and their cycling. Analysis of phospholipid fatty acid (PLFA) abundances and direct cell counts detected varying biomass that was not correlated with depth. Compound-specific carbon isotope analyses (δ13C and Δ14C) of the phospholipid fatty acids (PLFAs) and carbon substrates combined with genomic analyses did identify, however, distinct carbon sources and cycles between the two depth ranges studied. In the shallower boreholes at circa 1 kmbls, isotopic evidence indicated microbial incorporation of biogenic \{CH4\} by the in situ microbial community. At the shallowest site, 1.05 kmbls in Driefontein mine, this process clearly dominated the isotopic signal. At slightly deeper depths, 1.34 kmbls in Beatrix mine, the isotopic data indicated the incorporation of both biogenic \{CH4\} and dissolved inorganic carbon (DIC) derived from \{CH4\} oxidation. In both of these cases, molecular genetic analysis indicated that methanogenic and methanotrophic organisms together comprised a small component (<5%) of the microbial community. Thus, it appears that a relatively minor component of the prokaryotic community is supporting a much larger overall bacterial community in these samples. In the samples collected from >3 kmbls in Tau Tona mine (TT107, \{TT109\} Bh2), the \{CH4\} had an isotopic signature suggesting a predominantly abiogenic origin with minor inputs from microbial methanogenesis. In these samples, the isotopic enrichments (δ13C and Δ14C) of the \{PLFAs\} relative to \{CH4\} were consistent with little incorporation of \{CH4\} into the biomass. The most 13C-enriched \{PLFAs\} were observed in \{TT107\} where the dominant CO2-fixation pathway was the acetyl-CoA pathway by non-acetogenic bacteria. The differences in the δ13C of the \{PLFAs\} and the \{DIC\} and \{DOC\} for \{TT109\} Bh2 were ∼−24‰ and 0‰, respectively. The dominant CO2-fixation pathways were 3-HP/4-HB cycle > acetyl-CoA pathway > reductive pentose phosphate cycle. VL - 173 UR - http://www.sciencedirect.com/science/article/pii/S0016703715005852 ER -