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Abstract Radiocarbon (∆14C) measurements of nonstructural carbon enable inference on the age and turnover time of stored photosynthate (e.g., sugars, starch), of which the largest pool in trees resides in the main bole. Because of potential issues with extraction-based methods, we introduce an incubation method to capture the ∆14C of nonstructural carbon via respired CO2. In this study, we compared the ∆14C obtained from these incubations with ∆14C from a well-established extraction method, using increment cores from a mature trembling aspen (Populus tremuloides Michx). To understand any potential ∆14C disagreement, the yields from both methods were also benchmarked against the phenol-sulfuric acid concentration assay. We found incubations captured less than 100% of measured sugar and starch carbon, with recovery ranging from ~ 3% in heartwood to 85% in shallow sapwood. However, extractions universally over-yielded (mean 273 ± 101% expected sugar carbon; as high as 480%), where sugars represented less than half of extracted soluble carbon, indicating very poor specificity. Although the separation of soluble and insoluble nonstructural carbon is ostensibly a strength of extraction-based methods, there was also evidence of poor separation of these two fractions in extractions. The ∆14C of respired CO2 and ∆14C from extractions were similar in the sapwood, whereas extractions resulted in comparatively higher ∆14C (older carbon) in heartwood and bark. Because yield and ∆14C discrepancies were largest in old tissues, incubations may better capture the ∆14C of nonstructural carbon that is actually metabolically available. That is, we suggest extractions include metabolically irrelevant carbon from dead tissues or cells, as well as carbon that is neither sugar nor starch. In contrast, nonstructural carbon captured by extractions must be respired to be measured. We thus suggest incubations of live tissues are a potentially viable, inexpensive and versatile method to study the ∆14C of metabolically relevant (available) nonstructural carbon. 

Summary Shifts in the age or turnover time of non‐structural carbohydrates (NSC) may underlie changes in tree growth under long‐term increases in drought stress associated with climate change. But NSC responses to drought are challenging to quantify, due in part to large NSC stores in trees and subsequently long response times of NSC to climate variation.We measured NSC age (Δ¹⁴C) along with a suite of ecophysiological metrics inPinus edulistrees experiencing either extreme short‐term drought (−90% ambient precipitation plot, 2020–2021) or a decade of severe drought (−45% plot, 2010–2021). We tested the hypothesis that carbon starvation – consumption exceeding synthesis and storage – increases the age of sapwood NSC.One year of extreme drought had no impact on NSC pool size or age, despite significant reductions in predawn water potential, photosynthetic rates/capacity, and twig and needle growth. By contrast, long‐term drought halved the age of the sapwood NSC pool, coupled with reductions in sapwood starch concentrations (−75%), basal area increment (−39%), and bole respiration rates (−28%).Our results suggest carbon starvation takes time, as tree carbon reserves appear resilient to extreme disturbance in the short term. However, after a decade of drought, trees apparently consumed old stored NSC to support metabolism.