Tremella fuciformis is a dimorphic fungus that can undertake the reversible transition between yeast-like spores and hypha forms. In this project, we attempted to explore the differential proteins profile of dikaryotic yeast-like spores (FBMDS) and dikaryotic mycelium (DM) by applying the HRMS1-DIA full proteomics and PRM target proteomics synthetically. The results showed that a total of 5687 proteins were quantified, and 2220 of them (39.01%) showed more than a two-fold change in expression. The functional analysis of differential expression proteins (DEPs) confirmed that the DEPs were mainly located on membrane and nucleus, and FBMDS tended to express proteins involved in DNA replication and transcription, DNA damage repair, biosynthesis, and metabolism. At the same time, DM exhibited increased expression of signal transduction such as MAPK signaling pathway, Ras signaling pathway.