There is increasing interest in perennial crops to build soil carbon (C), but the mechanisms underlying soil C accrual in perennial croplands remain unclear, especially over time in the first years of perennial crop growth. To address this gap, research is needed that directly tracks intra-annual C fluxes through crop-microbial-soil pools, evaluating the capacity of perennial crops to build soil C over intra-decadal time periods. We conducted a 13C isotope-tracer study to compare within-season C uptake and crop-microbial-soil C partitioning patterns between 1-year-old (IWG-1) and 2-year-old (IWG-2) stands of a novel perennial grain crop, intermediate wheatgrass (IWG; Thinopyrum intermedium (Host) Barkworth and Dewey). We compared these to a common annual grain crop, spring wheat (Triticum aestivum L.). Crop shoots, roots, soil, and soil respired-C were sampled ten times over a 90-day chase period. We also measured the incorporation of recently assimilated 13C into soil microbial biomass (13C PLFA) and functional groups over the first 7 days post-label application. Overall, IWG-1 assimilated almost 1670 mg 13C m−2 during the study period, nearly twice that of IWG-2 or wheat, but neither IWG system retained significant amounts of new C in soil. Rather, a higher proportion of assimilated new C was retained in IWG-1 in root tissues (14%) and arbuscular mycorrhizal fungi when compared to other cropping systems, while IWG-2 retained almost 50% of total assimilated C in aboveground crop tissues. We expect the shift from new C retention in belowground root-mycorrhizal networks to aboveground tissues is associated with a shift from an acquisitive to conservative growth strategy that occurs between the first and second IWG production years. The observed shift in C partitioning patterns and potential change in growth strategy limited the allocation and retention of new C in soil as IWG aged, adding valuable context to our understanding of why perennial grain crop establishment seldom leads to significant carbon gains in the first several years following establishment.