In this study, Loropetalum chinense var. rubrum was used as tested material. A flavonol synthase (FLS) homology gene named LcFLS1 was cloned by PCR based on the transcriptome sequencing results. The bioinformatic analysis shows that the open reading frame of LcFLS1 gene contains 996 base pairs encoding 331 amino acids. Amino acid alignment results indicate that LcFLS1 may encodes a typical 2oxoglutarate and Fe(II)dependent oxygenase. Protein structure prediction indicates there are ten loci at the core domain of globular protein interacting with 2oxoglutarate ligand in LcFLS1. The phylogenetic tree analysis illustrates that LcFLS1 is closely related to Camellia sinensis and other woody plants but is far related to herbaceous plants like Arabidopsis thaliana. The express pattern of different tissues by qPCR shows that LcFLS1 expressed highest in L. chinense var. rubrum flower while lowest in stem. The pLcFLS1SUPER1300 overexpression vector was successfully constructed and transferred into Arabidopsis thaliana by Agrobacterium infection inflorescence method to obtain transgenic plants. PCR identification shows that positive transgenic plants of LcFLS1 were obtained. This research laid important foundations for studying the mechanism of flavonol biosynthesis as well as development and utilization of medicine value in L. chinense var. rubrum.