Analysis of Microbial Community Structure and Cultivation Performance Assessment in Tremella fuciformis Liquid Inoculum

Liquid inoculum is widely adopted in the mushroom industry, yet preparing <i>Tremella fuciformis</i> liquid inoculum remains challenging due to its complex microbial community and dimorphic growth. This study aimed to establish a reliable protocol for <i>T. fuciformis</i> liquid inoculum and assess its practical application. Initially, liquefied spawn was produced by liquefying solid spawn. The application of standard liquefied spawn increased fruiting body yield by 8.2% (502.4 g/kg dry substrate) compared to solid spawn, but exhibited substantial batch-to-batch variation due to unstable microbial communities and low <i>Tremellomycetes</i> abundance. To address these limitations, liquid spawn was developed via pre-culture of pure <i>T. fuciformis</i> and <i>Annulohypoxylon stygium</i> mycelia. Cultivation tests demonstrated significantly enhanced performance with 608.2 g of fruiting bodies, which represented 11.1% improvement compared to solid spawn. Moreover, consistent yields could be observed across multiple batches. This stability was attributed to stable microbial community structure and the dominance of <i>Tremellomycetes</i> (abundance > 50%) in the fungal community. These results confirm the cultivation performance of <i>T. fuciformis</i> liquid spawn, highlighting its potential as an effective alternative to solid spawn for <i>T. fuciformis</i> industrial production.