Chemotype differentiation and environmental drivers of bioactive metabolites in Cordyceps chanhua from diverse geographical origins

Two chemotypes, CTA and CTB, were identified in Cordyceps chanhua for the first time. • Intracellular nucleosides were key markers of chemotype differentiation. • CTA showed significantly higher intracellular uridine, guanosine, adenosine, and HEA than CTB. • Altitude and precipitation were the major environmental factors associated with chemotype differentiation. • Intracellular nucleosides showed significant negative correlations with altitude and precipitation and strong positive correlations with each other. Cordyceps chanhua is a valuable medicinal and edible fungus with great potential for functional food development because of its bioactive metabolites with significant pharmacological activities. However, samples from different geographical origins show significant variations in metabolite composition and content. Existing studies are mostly limited to local regions and lack systematic evaluation based on multi-regional samples, particularly regarding chemotype classification and its environmental driving mechanisms. Therefore, this study systematically analyzed the differences in bioactive metabolites, chemotype differentiation patterns, and their environmental driving factors using multi-regional samples. A total of 123  C. chanhua samples were collected from 16 regions across 8 provinces in China. Key bioactive metabolites, including uridine, guanosine, adenosine, N6-(2-hydroxyethyl)-adenosine (HEA), cordycepic acid, extracellular polysaccharides, and ergosterol, were determined in liquid fermentation broth. The results showed significant geographical differences in metabolite accumulation. Hierarchical cluster analysis (HCA) and principal component analysis (PCA) classified the samples into two chemotypes, Chemotype A (CTA) and Chemotype B (CTB). CTA exhibited significantly higher intracellular nucleoside contents than CTB ( p  < 0.01), and intracellular nucleosides were identified as the core variables driving chemotype differentiation. Environmental analysis showed that CTA was mainly distributed in low-altitude and low-precipitation regions, while intracellular nucleosides were significantly negatively correlated with altitude and precipitation. Correlation network analysis further revealed synergistic accumulation patterns among multiple metabolites. This study revealed the geographical variation characteristics of bioactive metabolites in C. chanhua , identified its chemotypes for the first time, and clarified that intracellular nucleosides are key chemical markers for chemotype classification. Download: Download high-res image (340KB) Download: Download full-size image