Abstract: Four key enzyme genes (CHS, DFR, F3′5′H, ANS) involved in anthocyanins synthesis were cloned from the flower petals of Phalaenopsis with homology sequence cloning.The length of the sequence was 302bp, 275bp, 285bp and285bp, respectively.Sequence analysis showed that these protein shared high similarity to other proteins from Dendrobium, Cymbidiumet al, ranging from 81%-95%, 67%-91%, 68%-92%and 83%-89%, respectively.The results of phylogenetic analysis were in agreement with those described in plant taxonomy.Resultsof Real-time fluorescence quantitative PCR analysis showed that the relative expression level of CHSand F3′5′H gene was the highest in the flower large bud and early flowering stages, and then decreased in blooming.The expression level of CHSand F3′5′H gene in petals and lip was greater than in sepals, the trace expression revealed in Leaves and roots.