Poria cocos (syn. Wolfiporia cocos) is an obligate saprophytic fungus that can produce sclerotia only when cultivated on pine wood. Despite its wide cultivation, the nutritional adaptation of this fungus to pine substrate and the host defensive responses triggered during colonization remain poorly characterized. Elucidating how Poria cocos colonizes pine and triggers host defenses is critical to uncover its nutrient dependence. GO and KEGG analyses revealed that protein degradation, carbon metabolism, protein processing in the endoplasmic reticulum, and proteasome pathways were the major enriched pathways, indicating that a large amount of pine wood protein was degraded after Poria cocos colonization. Consistent with these omics signatures, the fungus acquired its primary nitrogen source via breakdown of pine structural and metabolic proteins, while pine carbohydrates acted as its main carbon source. Nutritional profiling confirmed the depletion of soluble protein and total sugars in pine wood, alongside elevated defensive polyphenols and flavonoids, reflecting an active stress response from viable pine parenchyma cells upon colonization. No significant shifts in mineral element concentrations were detected in colonized pine wood, indicating that Poria cocos selectively absorbs target minerals without altering the overall mineral pool of host wood tissue. Collectively, this work elucidates the core nutritional strategy of Poria cocos during pine colonization and identifies key molecular clues to advance optimized artificial cultivation, laying a solid theoretical foundation for revealing its obligate saprophytic lifestyle.