The buds of oriental hybrid lily ‘Sorbonne’ were used to understand the regulatory mechanisms of flower coloration. Based on previous transcriptome database, we successfully cloned the coding sequence and genome sequence of flavanone 3hydroxylase (F3H). The cDNA sequence had the encoding region of 1 110 bp, and is predicted to encode a protein with 369 amino acids; the DNA fulllength of 1 438 bp contained three exons and two introns, and is named LhSorF3H. Bioinformatics analysis results revealed that F3H was highly conservative in evolution and there is a high similarity between the proteins encoded by LhSorF3H and F3H of Tulipa fosteriana. Analysis by semiquantitative reverse transcriptionPCR (RTPCR) revealed that LhSorF3H gene showed the strongest expression in tepals, stigma and style, lower expression in filament, ovary, immaturestem, stemroot and upperleaf, the lowest expression in anther and lowerleaf of ‘Sorbonne’. LhSorF3H expressed in almost every stage except S2 and S6. In addition, dark treatment can reduce the expression of LhSorF3H, and it showed a lowest expression after two hour dark treatment, and after that, the expression started to go up. With the increasing of illumination time, the expression level of LhSorF3H began to increase slightly, but there was no significant difference between the light time groups.