Laboratory and in-situ insights into the chemoautotrophic and zero-valent sulfur formation pathway in a cold-seep bacterium

Microbial sulfur cycling and chemoautotrophy are focal points of research in cold seeps. However, limited culture-dependent and in-situ studies have described the biological features and ecological significance of chemoautotrophic sulfur-oxidizing bacteria. In this study, we isolated Guyparkeria hydrothermalis SP2, a thiosulfate/sulfide-oxidizing chemoautotrophic bacterium, from cold-seep sediment. Electron microscopy, Raman spectroscopy, and stoichiometry confirmed the efficient production of zero-valent sulfur (ZVS) by G. hydrothermalis SP2. Genomic, transcriptomic, and qRT-PCR analyses revealed its utilization of the Sox pathway for thiosulfate oxidation and the fccB gene for sulfide oxidation. Its chemoautotrophic capability mediated by the Calvin-Benson-Bassham (CBB) cycle was identified through isotopic and qRT-PCR analyses. In-situ studies demonstrated its ability to produce ZVS by oxidizing sulfide in cold seeps, with a preference for different genes compared to those under laboratory conditions. Metagenomic and metatranscriptomic analyses indicated the ubiquity of its sulfur oxidation-based chemoautotrophic pathway in cold seep sediments. Therefore, this strain holds significance for investigating sulfur oxidation-based chemoautotrophic pathways in cold seeps.