Purpose <p><i>Curculigo orchioides</i> (Hypoxidaceae) is widely used in traditional medicine due to its antioxidant, antidiabetic, antiosteoporotic, and neuroprotective properties. This study presents, for the first time, a comparative histological and histochemical analysis of <i>in vitro</i>–derived propagules using a simple and reliable approach.</p> Methods <p>Histological observations were performed on cross-sections of callus tissue and plantlet organs (leaves, petioles, and roots). Histochemical tests were conducted to evaluate whether <i>in vitro</i> propagules produce specialized metabolites comparable to those found in the organs of the mother plant. Secretory structures in <i>in vitro</i> propagules were identified from razor-blade sections prepared according to standard fixation and light-microscopy procedures. Various histochemical reagents were applied to detect different classes of metabolites.</p> Results <p><i>In vitro</i> propagules exhibited secretory cavities and idioblasts, which served as sites for the localization of specialized metabolites. The formation of secretory cavities was initiated at the callus stage. Histochemical analyses showed positive reactions exclusively for phenolic compounds in idioblasts, secretory cavities, and organogenic callus tissues.</p> Conclusion <p>These results demonstrate that organogenic callus and <i>in vitro</i>–derived plantlet organs of <i>C. orchioides</i> are capable of producing phenolic compounds under controlled culture conditions.</p> Graphical Abstract <p></p>