The cut flowers of Paeonia suffruticosa ‘ Luoyang Hong ’ were used as materials. Effects of different temperature treatments (22℃,15℃ and 4℃) on flower color and anthocyanin biosynthesis were researched. The results showed that : compeared with 22℃, flowers under 15℃ and 4℃ treatments showed lower lightness, higher redness, higher chroma and increased anthocyanin accumulation. Color quality is obviously better than that of 22℃ treatment. qRT-PCR was used to analyze the relative expression levels of anthocyanin biosynthetic genes and results showed that 15℃ and 4℃ treatments promoted anthocyanin biosynthesis in the cut flowers via the promotion of related genes transcription (PsCHS1, PsCHI1, PsF3’H1, PsANS1, PsDFR1, PsMYB2, PsbHLH1 and PsbHLH3). Low-temperature mainly regulated and controled anthocyanin biosynthesis by upstream gene PsCHS1 and PsCHI1. Downstream genes PsDFR1, PsANS1 and PsF3’H1 were sensitive to low-temperature, which have the biggest increase transcription by 4℃ treatment. The above mentioned structural and regulatory genes were the important key of the anthocyanin biosynthesis.