The regulatory role of plant carbohydrate status and root exudation on soil CO<sub>2</sub> efflux has been demonstrated, yet the underlying mechanisms, particularly through root respiration, remain largely theoretical. In this study, we analyzed the cospectral variation of soil autotrophic (Ra) and heterotrophic (Rh) respiration components with key physiological and environmental factors, including gross primary productivity (GPP), photosynthetically active radiations (PAR), soil temperature (Ts) and volumetric water content (VWC), to evaluate their relative contributions in a subtropical mature shortleaf pine forest in the southern United States. The findings reveal a strong diurnal relationship between Rh and both GPP and PAR, in contrast to the weaker and more variable associations observed with Ra. This suggests that substrate availability was a key limitation of Rh on a diurnal basis, and that recently assimilated carbohydrates were directly discharged into the soil via root and mycorrhizal exudates. The consistent 2–4 hour time lag between Rh relative to GPP is consistent with the propagation rate of phloem pressure-concentration waves. While a diurnal peak in Rh-Ts covariance was also detected, the time lag of Rh in relation to Ts varied between positive and negative values, precluding this from being a causal relationship. Ra had a similarly strong cospectral peak with GPP as Rh, but with inconsistent lag, likely because of carbon availability from local starch reserves.