An understanding of ecosystem-level carbon (C) sequestration, or net ecosystem production (NEP), requires the separation of heterotrophic, microbial respiration (RH) from autotrophic, root-derived respiration (RA) as the components of RS (i.e., NEP = NPP - RH). However, separating these two sources in situ has been problematic since they are closely coupled. This study utilizes two similarly aged Pinus taeda L. stands, 8 and 9 years-old, aimed at quantifying these two respiration components through in-situ root severing. In order to use root-severing treatments to separate RS into RH and RA components, confirmation of carbohydrate depletion coupled to RA decline is crucial. This study evaluated the changes in CO2 flux rates and carbohydrate supply upon root severing in Pinus taeda L. using a controlled laboratory validating a two-part field study. The first field study used root-severing cores to test in-situ if respiration components can be attained based on the depletion of carbohydrate supply. The second field study was aimed at how future changes in climate might affect the ability of forests to store C and how modern forestry practices might affect changes and was conducted over the course of two installations, spring and summer 2012. In this study we examined the effects of fertilization (0 and 100.9 kg N ha-1 ) and throughfall reduction (0 and -30%) on total soil respiration (RS) as well as the heterotrophic contribution to RS, in a fully replicated (n=4), 2x2 factorial design. In the controlled lab experiment RS and RA declined by 86% and 95% respectively by the end of an 86 day trial and NSC carbohydrates declined by 60% for soluble, 29% for insoluble, and 43% for total (soluble + insoluble). The decline of RA was highly correlated to with the decline of NSC’s at 0.90, 0.69 and 0.93 for soluble, insoluble and total, respectively. The companion field study revealed a mean decrease 21±0.5% of over the final three dates when severed root respiration stabilized. In the second study, testing throughfall reduction and fertilization levels there were no fertilization by throughfall reduction interactions on the contribution of RH to RS in either the spring or summer; however, the main effect of throughfall reduction was significant in the spring. During the spring, the mean contribution of RH to RS for ambient throughfall plots was 96±6.4%, while the mean contribution under throughfall reduction was 68±1.9%. During the summer, there were no differences among treatments and the overall contribution of RH to RS was 78±1.6%. Collectively, both of these studies revealed that the severing of roots from their primary energy source and the subsequent depletion of stored NSC that the use of in-situ methods allows for the quantification of soil respiration components RA and RH. Using these estimates to model NEP in the short-term can be variable by season, however, long-term monitoring may simplify future NEP modeling scenarios