Shiyong Wen, Wenjiao Li, Ruizhen Jian, Xunfan Wei, Xiuli Sun, Cheng Liu, Wen Li, Cuiping Wen, Wen Yang, Yifan Yang, Huisong Wang, Zhennan Qiu
Among the concentrations tested, 50 mg/L ASAE produced the greatest biomass accumulation at day 7, increasing mycelial dry weight by 22.7% relative to the untreated control, whereas 1,000 mg/L ASAE inhibited biomass accumulation. However, the treatment effect was no longer statistically significant at day 9, indicating a transient acceleration of biomass accumulation rather than an increase in final biomass yield under the conditions tested. Two-way ANOVA identified significant effects of cultivation time, treatment, and their interaction on mycelial dry weight. WGCNA module eigengenes were further analyzed using models incorporating cultivation time, treatment, and their interaction. MEblue remained significantly associated with all three factors, including their interaction, but was interpreted as a time- and treatment-associated co-expression module rather than a treatment-specific regulatory module. Direction-separated GSEA showed time-resolved enrichment of gene sets related to energy metabolism, RNA processing, protein homeostasis, carbon metabolism, translation, intracellular trafficking, and carbohydrate-associated metabolism. Upregulated GSEA leading-edge genes were intersected with MEblue genes for candidate-gene visualization without performing a second enrichment analysis. Independent RT-qPCR experiments confirmed the expression trends of selected candidates, including CHSG, UAP1, ALG5, and RPN12. A homology-based transcript-collapse sensitivity analysis produced qualitatively similar module- and pathway-level patterns.
INTRODUCTION: Lyophyllum decastes is an edible mushroom with nutritional and medicinal value, but approaches for accelerating mycelial biomass accumulation during submerged cultivation remain limited.
METHODS: This study evaluated the effects of Acanthopanax senticosus aqueous extract (ASAE) on submerged mycelial growth and examined associated transcriptional responses using time-course RNA sequencing, weighted gene co-expression network analysis, gene set enrichment analysis, and independent RT-qPCR experiments.
RESULTS: Among the concentrations tested, 50 mg/L ASAE produced the greatest biomass accumulation at day 7, increasing mycelial dry weight by 22.7% relative to the untreated control, whereas 1,000 mg/L ASAE inhibited biomass accumulation. However, the treatment effect was no longer statistically significant at day 9, indicating a transient acceleration of biomass accumulation rather than an increase in final biomass yield under the conditions tested. Two-way ANOVA identified significant effects of cultivation time, treatment, and their interaction on mycelial dry weight. WGCNA module eigengenes were further analyzed using models incorporating cultivation time, treatment, and their interaction. MEblue remained significantly associated with all three factors, including their interaction, but was interpreted as a time- and treatment-associated co-expression module rather than a treatment-specific regulatory module. Direction-separated GSEA showed time-resolved enrichment of gene sets related to energy metabolism, RNA processing, protein homeostasis, carbon metabolism, translation, intracellular trafficking, and carbohydrate-associated metabolism. Upregulated GSEA leading-edge genes were intersected with MEblue genes for candidate-gene visualization without performing a second enrichment analysis. Independent RT-qPCR experiments confirmed the expression trends of selected candidates, including CHSG, UAP1, ALG5, and RPN12. A homology-based transcript-collapse sensitivity analysis produced qualitatively similar module- and pathway-level patterns.
DISCUSSION: These findings indicate that ASAE-associated acceleration of L. decastes biomass accumulation is accompanied by time-resolved transcriptional remodeling. The proposed biological interpretation remains hypothesis-generating, and direct physiological, biochemical, protein-level, and genetic validation will be required to establish causality.