Genetic legacy of the deep subsurface recorded in the outflow channel of a terrestrial serpentinizing seep (Luzon, the Philippines)

Abstract:

The deep subsurface microbial community represents the largest biome on Earth, yet accessing this deep biosphere is challenging. Fluids seep along fractures from aquifers that may support diverse microbial communities, living off hydrogen gas generated by radiolysis, serpentinization, or thermogenic reactions. A serpentinizing seep, emanating fluids as high as pH 11.27, was found to accrete meters-long carbonate terraces in the Zambales ophiolite range (Luzon, the Philippines). Samples were collected at several locations along the Poon Bato (PB) River, focusing primarily on the pools and terraces formed by carbonate rimstone (Figure 1). As serpentinizing fluids are exposed to the atmosphere, dynamic niches are established in surface sediments. We propose that the high pH, reducing, high Ca⁺² fluid pool terraces reflect remnants of deep subsurface microbial communities, based on high-throughput 16S rRNA sequencing data. In total, eight samples were collected for environmental DNA analysis. Post-sequence analysis revealed a total of 927126 counts, with an average of 115890.75 counts per sample. Many taxa aligned with cultured representatives of serpentinizing seep-associated taxa, including Bacteroidetes, Clostrida, Chloroflexi, Methylococcales, and Xanthomonadales.

Geochemical data indicates an average fluid temperature of 28.9°C, and pH that varies from 9.22-11.27. Total carbon wt.% of solids was highest in a shallow pool shaped by boulders, where calcite precipitation occurred over nearly every surface. Dissolved oxygen (DO%) was highest at PB1 main pool (60%), although a calcite skin had formed along the air-water interface. Dissolved inorganic carbon (DIC) at PB1 main pool was 1.3 ppm, while at PB2 main pool, the DIC was higher (6.0 ppm). The lack of calcite skin may allow more direct access to atmospheric carbon dioxide at PB2. The isotopic value of carbon-13 was depleted at PB1 relative to PB2 (δ¹³C VPDB -25.4 ‰ versus δ¹³C VPDB ‰ -17.5, respectively). The DOC concentration at PB1 main pool was 0.3 ppm and 1.15 ppm at PB2. Given the low DIC concentrations at PB1, it is suggested that heterotrophy may dominate over autotrophy in the system. This suggests that the highly reducing, high pH fluids emanating from fluid seeps at Poon Bato influence surface communities via inundation with serpentinizing fluid.