| Summary: | Abstract Altogether, 14 basidiomycetes (12BAD, 95PCH, 9WCOC, 5PSA, 96BCI, 331SHIBD, 4MSC, 74HFA, 220MPS, 115PFLA, 111 ICO C, 16LED, 6TSU, and 61LYP) were grown on solid and in liquid media using hairy roots of genetically modified Hypericum perforatum (L.) as the only source of carbon and nitrogen. After the first screening by GC‐MS/MS‐O, two fungi (115PFLA and 61LYP) which resulted in the most pleasant complex natural flavor by biotransformation were selected for further analysis. Twenty‐four new volatile compounds were produced, from which 21 were identified (ethyl hexanoate, ethyl octanoate, benzaldehyde, 2‐undecanone, (E,E)‐2,4‐decadienal, 1‐octen‐3‐one, (E)‐2‐nonenal, ethyl nonanoate, 2‐heptenal, 1‐methoxy‐4‐methylbenzene, 3‐octanone, 1‐decen‐3‐one, (E)‐2‐octenal, 1‐octen‐3‐ol, β‐linalool, ±trans‐nerolidol, anisole, methyl benzoate, 2‐pentylfuran, 1,3‐dichloro‐2‐methoxybenzene, and 1‐dodecanol). Thereof, 15 compounds were perceived at the ODP, from which 13 were identified. Compound identification was performed by comparison of Kovats indices (KI) and mass spectra to those of authentic reference compounds on a polar VF‐WAXms column using headspace solid‐phase microextraction–gas chromatography–mass spectrometry (HS‐SPME‐GC‐MS).
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