The flavonoids content determines the quality characteristics of Ginkgo biloba extract that could be increased by using of plant growth regulators. The objective of study was to investigate the effect of chlorocholine chloride (CCC), an anti-gibberellin growth retardant, on photosynthesis, chlorophyll, soluble sugar, total amino acids and phenylalanine contents, flavonoid accumulation, and flavonoids enzyme activity in G. biloba leaves. The ginkgo seedlings were grown in the greenhouse conditions with foliar applications of 0 (control), 0.5, 1.0 and 2.0 g l-1 CCC. Results showed that 0.5, 1.0 and 2.0 g l-1 CCC treatments significantly increased photosynthetic rates of leaves, the contents of chlorophyll, soluble sugar, total amino acids and phenylalnine in ginkgo leaves. Total polyphenols, flavonoids, anthocyanins content, phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS) and chalcone isomerase (CHI) activities were all significantly increased by 1.0 and 2.0 g l-1 CCC treatments. Foliar treatment with CCC therefore might be a useful means of improving pharmacological properties of G. biloba leaves.