Raman microspectroscopy for in situ examination of carbon-microbe-mineral interactions

The changing paradigm of soil organic matter formation and turnover is focused at the nexus of microbe-carbon-mineral interactions. However, visualizing biotic and abiotic stabilization of C on mineral surfaces is difficult given our current techniques. Therefore we investigated Raman microspectroscopy as a potential tool to examine microbially mediated organo-mineral associations. Raman microspectroscopy is a non-destructive technique that has been used to identify microorganisms and minerals, and to quantify microbial assimilation of 13C labeled substrates in culture. We developed a partial least squares regression (PLSR) model to accurately quantify (within 5%) adsorption of four model 12C substrates (glucose, glutamic acid, oxalic acid, p-hydroxybenzoic acid) on a range of soil minerals. We also developed a PLSR model to quantify the incorporation of 13C into E. coli cells. Using these two models, along with measures of the 13C content of respired CO2, we determined the allocation of glucose-derived C into mineral-associated microbial biomass and respired CO2 in situ and through time. We observed progressive 13C enrichment of microbial biomass with incubation time, as well as 13C enrichment of CO2 indicating preferential decomposition of glucose-derived C. We will also present results on the application of our in situ chamber to quantify the formation of organo-mineral associations under both abiotic and biotic conditions with a variety of C and mineral substrates, as well as the rate of turnover and stabilization of microbial residues. Application of Raman microspectroscopy to microbial-mineral interactions represents a novel method to quantify microbial transformation of C substrates and subsequent mineral stabilization without destructive sampling, and has the potential to provide new insights to our conceptual understanding of carbon-microbe-mineral interactions.