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<i>Antrodia cinnamomea</i> is a valuable edible and medicinal mushroom with antitumor, hepatoprotective, and antiviral effects that play a role in intestinal flora regulation. Spore-inoculation submerged fermentation has become the most efficient and well-known artificial culture process for <i>A. cinnamomea</i>. In this study, a specific low-molecular compound named 1,8-cineole (cineole) from <i>Cinnamomum kanehirae</i> Hay was first reported to have remarkably promoted the asexual sporulation of <i>A. cinnamomea</i> in submerged fermentation (AcSmF). Then, RNA sequencing, real-time quantitative PCR, and a literature review were performed to predict the molecular regulatory mechanisms underlying the cineole-promoted sporulation of AcSmF. The available evidence supports the hypothesis that after receiving the signal of cineole through cell receptors Wsc1 and Mid2, Pkc1 promoted the expression levels of <i>rlm1</i> and <i>wetA</i> and facilitated their transfer to the cell wall integrity (CWI) signal pathway, and <i>wetA</i> in turn promoted the sporulation of AcSmF. Moreover, cineole changed the membrane functional state of the <i>A. cinnamomea</i> cell and thus activated the heat stress response by the CWI pathway. Then, heat shock protein 90 and its chaperone Cdc37 promoted the expression of <i>stuA</i> and <i>brlA,</i> thus promoting sporulation of AcSmF. In addition, cineole promoted the expression of <i>areA</i>, <i>flbA</i>, and <i>flbD</i> through the transcription factor NCP1 and inhibited the expression of <i>pkaA</i> through the ammonium permease of MEP, finally promoting the sporulation of AcSmF. This study may improve the efficiency of the inoculum (spores) preparation of AcSmF and thereby enhance the production benefits of <i>A. cinnamomea</i>.