Although bioluminescence is documented both anecdotally and experimentally, the parameters involved in the production of fungal bioluminescence during wood colonization have not been identified to date. Here, for the first time, this work develops a methodology to produce a hybrid living material by manipulating wood colonization through merging the living fungus Desarmillaria tabescens with nonliving balsa (Ochroma pyramidale) wood to achieve and control the autonomous emission of bioluminescence. The hybrid material with the highest bioluminescence is produced by soaking the wood blocks before co-cultivating them with the fungus for 3 months. Regardless of the incubation period, the strongest bioluminescence is evident from balsa wood blocks with a moisture content of 700–1200%, highlighting the fundamental role of moisture content for bioluminescence production. Further characterization reveals that D. tabescens preferentially degraded hemicelluloses and lignin in balsa wood. Fourier-transform infrared spectroscopy reveals a decrease in lignin, while X-ray diffraction analysis confirms that the cellulose crystalline structure is not altered during the colonization process. This information will enable the design of ad-hoc synthetic materials that use fungi as tools to maximize bioluminescence production, paving the way for an innovative hybrid material that could find application in the sustainable production of light.