Despite the importance of soil organic matter (SOM) in C storage and provision of ecosystem services, the magnitude and direction of the response of SOM to climate change remains debated. Particularly contested is the role of biochemical recalcitrance in determining the biological stability of SOM, which in turn, may also vary with climate. Employing a climosequence study design controlling for confounding pedogenic factors, the research described in this thesis aimed to uncover the response of both SOM chemistry and SOM biological stability to changes in climate and associated land use shifts at the grassland-forest ecotone in west-central Saskatchewan. Characterization of SOM chemistry was achieved using two advanced analytical techniques: X-ray absorption near edge structure (XANES) spectroscopy and pyrolysis-field ionization mass spectrometry (Py-FIMS). Agreements between XANES and Py-FIMS revealed only minor differences in SOM chemistry resulting from a 0.7 °C mean annual temperature (MAT) gradient and associated broad differences in land use, but revealed a clear influence of depth within soil profiles. In contrast, long-term aerobic incubations revealed that biological stability of SOM varied with both climate and climate-induced differences in land use, but was not largely influenced by depth. Together, these findings suggest a decoupling of SOM chemistry and its biological stability, indicating that factors other than biochemical recalcitrance are the primary drivers of SOM persistence in these soils.