Ganoderma triterpenoids (GTs), a class of major active constituents of Ganoderma fungi, possess diverse structures and remarkable activities. In the present study, nine new GTs, namely applanoids A—I (1—9), were isolated from the medicinal fungus of Ganoderma applanatum. Their structures including absolute configurations were established by comprehensive spectroscopic analyses and ECD calculation. Applanoids A—E (1—5) represent the first example of GTs with 6/6/5/6/5 pentacyclic system and the formation of the ether ring between C‐15 and C‐20 involves Michael addition reaction. Furthermore, compounds 1—8 were evaluated for their human pregnane X receptor (hPXR) agonistic activity using dual‐luciferase reporter gene assay, and the results showed that compounds 1, 2 and 4 can dose‐dependently activate hPXR. This investigation further illustrated the structural diversity of GTs and provided new insights for searching PXR agonists from GTs.