The Metabolic Regulatory Mechanisms of Umami Amino Acids in Stropharia rugosoannulata

<i>Stropharia rugosoannulata</i> is a widely cultivated edible mushroom known for its nutritional value and umami flavour. Electronic tongue technology and metabolomics revealed that glutamic acid (Glu) and aspartic acid (Asp) levels were positively correlated with umami in the fruiting body developmental stages. Subsequent investigations found that overexpression of <i>SrCS</i> within the TCA cycle resulted in decreased levels of Glu and Asp. Integrating TF-gene-metabolite network modelling with experiments identified SrELT1 as a transcriptional regulator of <i>SrCS</i>. Different temperatures, cultivation substrates and genetics significantly impacted <i>SrELT1</i> and <i>SrCS</i> expression, thereby affecting Glu and Asp synthesis. The findings suggest that increased Citrate synthase (CS) activity channelled citrate into glycolysis and oxidative phosphorylation without excessive accumulation; in contrast, decreased CS activity shifted metabolism toward the production of metabolites like Glu and Asp. This study provides insights for enhancing the umami of <i>S. rugosoannulata</i>, thereby substantially increasing its market competitiveness in the premium food segment.