Abstract: Tissue separation is often used for purification and rejuvenation of edible fungi, but continuous tissue separation will lead to strain degradation. Using V844 as the initial strain T0, the subculture strain T1-T18 was obtained by continuous tissue isolation and subculture for 18 times. The physiological characteristics of T0-T18 and the enzyme activities related to cellulose, hemicellulose and lignin degradation were measured and analyzed by transcriptology. The results showed that with the increase of subculture time, the colony diameter of V. volvacea decreased gradually, and the mycelial growth rate and biological quality increased at first and then decreased. The activities of exoglucanase, laccase, manganese peroxidase and hemicellulase increased at first and then decreased. Compared with T0, those activities of T18 strain decreased by 34.2%, 71.1%, 90.0% and 58.7%, respectively. The activities of endoglucanase, filter paper enzyme and xylanase were not significantly different at T0-T4, and then began to decline. Compared with T0, T18 decreased by 66.7%, 86.4% and 67.9%, respectively. There was no significant difference in β-glucosidase activity (P>0.05). Transcriptomics analysis of T0, T4, T8, T12 and T16 showed that there were 34 differentially expressed genes related to matrix degradation enzymes (8 up-regulated,17 down-regulated, first up, then down 9). The physiological characters, related enzyme activities and transcription histology of the subculture of V. volvacea were analyzed synthetically. It was found that when the continuous tissue was separated and subcultured for four times, the strain of V. volvacea had obvious rejuvenation effect, but when the continuous subculture was more than 12 times, the tissue isolation and subculture would lead to the strain degradation.
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