Systematic investigations of the volatiles of field‐grown plants and shoot cultures of Gynura bicolor have been conducted to obtain indispensable plant physiological information for propagating plants of consistent quality. Volatiles obtained by solvent‐assisted flavour evaporation (SAFE) of solvent extracts from leaves of commercial cultivars, shoot cultures and their regenerates were analysed using gas chromatography (GC), GC–mass spectrometry (GC–MS) and GC–olfactometry (GC–O). It was revealed for the first time that this plant had the following chemical characteristics: field‐grown plants, i.e. those plants commercially cultivated in three different regions of Japan and 7 month‐old regenerates of shoot cultures had almost the same GC profiles; their major components were ( E )‐caryophyllene, α‐humulene and bicyclogermacrene; in contrast, the major components of the shoot cultures were ( Z , E)‐ α‐farnesene, ( E )‐caryophyllene and α‐copaene. The biosynthetic pathways for the sesquiterpenes found in these materials were presumed from the analytical results. Aroma evaluations using GC–O revealed that 1‐octen‐3‐one, ( Z )‐1,5‐octadien‐3‐one and trans ‐4,5‐epoxy‐( E )‐2‐decenal were aroma‐impact compounds contributing to the characteristic metallic, green aroma, especially in the shoot cultures. In addition to these results, it was found that 2‐isopropyl‐ and 2‐isobutyl‐3‐methoxypyrazines were additional compounds related to the green, earthy aroma in the regenerates. These aroma‐impact compounds were detected only because of the GC–O analysis. Copyright © 2009 John Wiley & Sons, Ltd.