Abstract
Amylostereum areolatum (Chaillet ex Fr.) Boidin (Russulales: Amylostereaceae) is a symbiotic fungus of Sirex noctilio Fabricius that has ecological significance. Terpenoids are key mediators in fungal-insect interactions, yet the biosynthetic mechanisms of terpenoids in this species remain unclear. Under nutritional conditions that mimic natural growth, A. areolatum was sampled during the lag phase (day 7), exponential phase (day 14), and stationary phase (day 21). Metabolome (solid-phase microextraction (SPME) combined with gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS)) and transcriptome (Illumina NovaSeq) profiles were integrated to investigate terpenoid-gene correlations. This analysis identified 103 terpenoids in A. areolatum, substantially expanding the known repertoire of terpenoid compounds in this species. Total terpenoid abundance progressively increased across three developmental stages, with triterpenoids and sesquiterpenoids demonstrating the highest diversity and abundance levels. Transcriptomic profiling (61.66 Gb clean data) revealed 26 terpenoid biosynthesis-associated genes, establishing a comprehensive transcriptional framework for fungal terpenoid metabolism. Among 11 differentially expressed genes (DEGs) (|log2Fold Change| ≥ 1, adjusted p < 0.05), HMGS1, HMGR2, and AaTPS1-3 emerged as key regulators potentially governing terpenoid biosynthesis. These findings provide foundational insights into the molecular mechanisms underlying terpenoid production in A. areolatum and related basidiomycetes.