Objective: Clarify the flavor characteristics of Tremella fuciformis cultivated on herbaceous substrate. Methods: In the study T. fuciformis cultivated on herbaceous substrate was used as the research object, while samples of yellow tremella and white tremella cultivated on conventional substrate were used as controls. The differences in flavor composition between them were analyzed using high-performance liquid chromatography and Headspace Solid Phase Microextraction Ggas Chromatography Mass Spectrometry (HS-SPME-GC-MS). And combined with intelligent sensory technology, its flavor characteristics were analyzed. Results: According to multi-dimensional evaluations such as fresh and sweet amino acid and flavor nucleotides content, taste activity value and equal freshness concentration value, and electronic tongue, the freshness intensity of this herb's tremella was lower than that of yellow tremella and white tremella. The results of HS-SPME-GC-MS analysis showed that 82 volatile compounds were identified in this herb Tremella, which were higher than those in white Tremella (71 species) and yellow Tremella (49 species). The main compounds were esters, accounting for 16.9%～22.4%, presenting a strong floral aroma. At the same time, 26 characteristic aroma components of Tremella, 9 unique components of white Tremella, and 4 unique components of this herb Tremella were identified; In terms of aroma substance content, this herb Tremella also holds an advantage. Conclusion: The traditional Chinese medicinal fungus has a light taste and a strong aroma, making it suitable for developing and preparing beverages, while yellow white fungus is more suitable as a dish.