The plant hormone auxin (indole‐3‐acetic acid, IAA) is involved in the control of many phenomena during plant development. By characterizing steady‐state free and conjugated IAA levels using a stable isotope dilution method coupled with gas chromatography‐ selected ion monitoring‐ mass spectrometry, this paper provides a detailed characterization of IAA metabolism in five liverworts, four mosses, and two tracheophytes. Long‐term IAA conjugation patterns were monitored by incubating actively growing tissue with 14C‐IAA and then analyzing the de novo synthesis of IAA conjugates with radioimaging techniques. The liverworts, mosses, and tracheophytes can be differentiated by the total amount of IAA metabolites, the proportion of free and conjugated IAA, the chemical nature of their IAA conjugates, and the rates of IAA conjugation. Our tentative conclusion is that the liverworts appear to employ a biosynthesis–degradation strategy for the regulation of free IAA levels, in contrast to the conjugation–hydrolysis strategy apparently used by the mosses and tracheophytes. Such alternative metabolic strategies may have profound implications for macroevolutionary processes in these plant groups.