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Summary Lignin is an important root chemical component that is widely used in biogeochemical models to predict root decomposition. Across ecological studies, lignin abundance has been characterized using both proximate and lignin‐specific methods, without much understanding of their comparability. This uncertainty in estimating lignin limits our ability to comprehend the mechanisms regulating root decomposition and to integrate lignin data for large‐scale syntheses.We compared five methods of estimating lignin abundance and composition in fine roots across 34 phylogenetically diverse tree species. We also assessed the feasibility of high‐throughput techniques for fast‐screening of root lignin.Although acid‐insoluble fraction (AIF) has been used to infer root lignin and decomposition, AIF‐defined lignin content was disconnected from the lignin abundance estimated by techniques that specifically measure lignin‐derived monomers. While lignin‐specific techniques indicated lignin contents of 2–10% (w/w) in roots, AIF‐defined lignin contents werec.5–10‐fold higher, and their interspecific variation was found to be largely unrelated to that determined using lignin‐specific techniques. High‐throughput pyrolysis–gas chromatography–mass spectrometry, when combined with quantitative modeling, accurately predicted lignin abundance and composition, highlighting its feasibility for quicker assessment of lignin in roots.We demonstrate that AIF should be interpreted separately from lignin in fine roots as its abundance is unrelated to that of lignin polymers. This study provides the basis for informed decision‐making with respect to lignin methodology in ecology. 

Summary Recent studies on fine root functional traits proposed a root economics hypothesis where adaptations associated with mycorrhizal dependency strongly influence the organization of root traits, forming a dominant axis of trait covariation unique to roots. This conclusion, however, is based on tradeoffs of a few widely studied root traits. It is unknown how other functional traits fit into this mycorrhizal‐collaboration gradient. Here, we provide a significant extension to the field of root ecology by examining how fine root secondary compounds coordinate with other root traits.We analyzed a dataset integrating compound‐specific chemistry, morphology and anatomy of fine roots and leaves from 34 temperate tree species spanning major angiosperm lineages.Our data uncovered previously undocumented coordination where root chemistry, morphology and anatomy covary with each other. This coordination, aligned with mycorrhizal colonization, reflects tradeoffs between chemical protection and mycorrhizal dependency, and provides mechanistic support for the mycorrhizal‐collaboration gradient. We also found remarkable phylogenetic structuring in root chemistry. These patterns were not mirrored by leaves. Furthermore, chemical protection was largely decoupled from the leaf economics spectrum.Our results unveil broad organization of root chemistry, demonstrate unique belowground adaptions, and suggest that root strategies and phylogeny could impact biogeochemical cycles through their links with root chemistry.