Although understanding the timing of the deposition of recent photosynthate into fine roots is critical for determining root lifespan and turnover using isotopic techniques, few studies have directly examined the deposition and subsequent age of root carbon. To gain a better understanding of the timing of the deposition of root carbon, we labeled four individual Sassafras albidum trees with 99% 13C CO2. We then tracked whether the label appeared in roots that were at least two weeks old and no longer elongating, at the time of labeling. We found that not only were the non-structural carbon pools (soluble sugars and starch) of existing first-order tree roots incorporating carbon from current photosynthate, but so were the structural components of the roots, even in roots that were more than one year old at the time of labeling.Our findings imply that carbon used in root structural and nonstructural pools is not derived solely from photosynthate at root initiation and have implications regarding the determination of root age and turnover using isotopic techniques.