Influence of Culture Conditions on the Growth of Callus and Content of Total Flavonoids in Tetrastigma Hemsleyanum

To set up the optimal culture conditions for the callus growth and the accumulation of total flavonoids,the influence of light,medium,plant growth regulators and organic additives on the growth of callus and content of total flavonoids in Tetrastigma hemsleyanum was analyzed.The callus growth and the accumulation of total flavonoids under different culture conditions(photoperiods,organic additives,basic meida,concentrations of 6-BA and NAA) were studied by single factor experiment and orthogonal test.For callus growth the optimum medium was 12 h/d photoperiods,MS+3mg/L 6-BA+2 mg/L NAA+2 g/L peptone;For accumulation of total flavonoids the optimum medium was 24 h/d photoperiods,B5+4 mg/L 6-BA+2 mg/L NAA+ 40 mg/L Phenylalanine.Callus of Tetrastigma hemsleyanum could be cultured by a Two-step Method: callus was cultured in the former medium for 30 days and then in the latter for 20 days.By using this method,total flavonoids in cultured callus of maximum concentration could reach 28.4±3.9 mg/g DW.Tetrastigma hemsleyanum grew significantly faster in callus culture.The total flavonoids content in callus was significantly higher than that of raw plant leaves,close to the rhizomes.