Physiological and Flavonoid Metabolic Responses of Black Locust Leaves to Drought Stress in the Loess Plateau of China

Drought threatens the stability of artificial black locust forests on the Loess Plateau, yet there is limited research on the physiological and metabolic responses of mature black locust to drought stress. This study employed a throughfall exclusion system—i.e., moderate drought (40% throughfall reduction), extreme drought (80% throughfall reduction), and 0% throughfall reduction for control—to analyze leaf microstructure, relative water content (RWC), osmotic adjustment substances, hormone levels, and flavonoid metabolites in black locust under controlled drought stress. The results demonstrated that as drought stress intensified, stomatal aperture and density decreased, while trichome density and length exhibited significant increases. MDA, proline, IAA, and osmotic adjustment substances (soluble protein, reducing sugar, and total sugar) first increased and then decreased as drought stress intensified. A total of 245 flavonoid compounds were identified through metabolomic analysis, among which 91 exhibited differential expression under drought treatments. Notably, 37 flavonoids, including flavonols and glycosylated derivatives, were consistently upregulated. These findings suggest that drought stress can lead to the accumulation of flavonoids. This study explored the physiological and metabolic responses of mature black locust trees to drought stress, offering insights for selecting drought-resistant species in vegetation restoration and informing ecological management practices in arid regions.