Regulation of flavonoid metabolism in ginkgo leaves in response to different day-night temperature combinations

Flavonoids are the most important secondary metabolites in ginkgo (Ginkgo biloba L.) leaves that determine its medicinal quality. Studies have suggested that secondary metabolism is strongly affected by temperature in other plant species, but little is known about ginkgo. In this study, we investigated the effects of different day-night temperature combinations (15/10, 25/20, and 35/30 °C (day/night)) on key enzyme activity, growth regulator concentrations, and flavonoid accumulation in ginkgo leaves. We found that phenylalanine ammonia-lyase (PAL) activity was enhanced and inhibited at 15/10 and 35/30 °C, respectively. Cinnamate-4-hydroxylase (C4H) activity was relatively stable under the three temperature conditions, and the p-coumarate CoA ligase (4CL) activity showed different trends under the three temperature conditions. The concentrations of flavonoid constituents (quercetin, kaempferol, and isorhamnetin) were decreased and increased under the 35/30 and 15/10 °C conditions, respectively. Low temperature promoted soluble sugar accumulation, while temperature had a limited impact on the accumulation of soluble protein. The pattern of change in the total flavonoid concentration was not always in agreement with PAL activity due to its complex pathway. Indoleacetic acid (IAA) and gibberellin (GA) changes shared similar patterns and had limited effects on flavonoid accumulation, while abscisic acid (ABA) acted as a promotor of flavonoid accumulation under high-temperature conditions. The total flavonoids achieved the highest content under the 15/10 °C treatment on the 40th day. Therefore, the lower temperature (15/10 °C) is more favorable for flavonoid accumulation and will provide a theoretical basis for further study.