Metabolomics-Based Analysis of the Growth-Promoting Function of Endophytic Fungi

Medicago sativa is one of the world’s most important forage plants, possessing strong nitrogen-fixing and regrowth capabilities. Promoting its growth not only enhances stress resistance but also reduces the use of chemical fertilizers. The value of Centella asiatica is primarily reflected in its medicinal properties. Currently, endophytic fungal resources of C. asiatica are scarce, and their potential to promote medicinal components and the underlying mechanisms remains unclear. This study employed DNA extraction techniques to isolate and identify endophytic fungi from different parts of C. asiatica. We systematically analyzed the plant growth-promoting traits of endophytic fungi. After screening for the optimal strain and inoculating it into Medicago sativa, we elucidated the mechanisms underlying its growth-promoting effect using metabolomic sequencing. Research findings: A total of 18 endophytic fungal strains were isolated, belonging to 12 genera. Among them, five indole-3-acetic acid (IAA) strains were identified, with strain J4 demonstrating the highest IAA production (17.157 mg·L−1). The J4 strain has iron-transporting carrier activity, while 15 strains exhibit nitrogen-fixing activity. Inoculation with the Plectosphaerella plurivora strain significantly increases M. sativa’s germination rate, fresh weight, dry weight, and plant height. Metabolomic analysis indicates that P. plurivora may promote anthocyanin and jasmonic acid accumulation by regulating pathways such as flavonoid biosynthesis and pyrimidine metabolism, thereby promoting growth. This study reveals the mechanism by which endophytic fungi enhance M. sativa growth at the metabolomic level. This study reveals the growth-promoting mechanism of endophytic fungi in M. sativa from a metabolomic perspective, providing a theoretical basis for increasing forage yield and offering new insights into sustainable agricultural development.