目的进一步优化芍药愈伤组织中积累芍药双苷(芍药苷和芍药内酯苷)的培养条件。方法以芍药愈伤组织中芍药双苷的量为响应值,采用Plackett-Burman实验设计对8个影响愈伤组织生长和芍药双苷积累的植物激素(NAA、2,4-D、6-BA、TDZ)的质量浓度、蔗糖的含有量、Ca2+量、NH4+∶NO3-的比例及光照中筛选出了3个关键因素,通过最陡爬坡逼近和响应面优化方法确定这些因素的最适水平。结果三个因素的最佳条件为:40.20 g/L蔗糖、0.33 mg/L 6-BA和1.08 mg/L NAA。结论在此最优培养条件下,芍药愈伤组织合成芍药双苷的能力比优化前提高了33%,芍药双苷占愈伤组织干重含有量达到37.60 mg/g。
AIM To optimize key factors in callus culture of Paeonia lactiflora Pall. for accumulation increase of paeoniflorin and albiflorin. METHODS With the total amount of paeoniflorin and albiflorin as the response indicator,three out of eight parameters for the measurement of callus culture of Paeonia lactiflora Pall.,phytohormone( NAA、2,4-D、6-BA、TDZ),sucrose,Ca2 +,ratio of NH4 +and NO3-,and light treatment were checked by Plackett-Burman experimental design. The steepest ascent method in combination with central composite design and response surface method were applied to determining the optimal levels of these factors. RESULTS 40. 20 g / L sucrose,0. 33 mg / L 6-BA,and 1. 08 mg / L NAA were identified as the most satisfactory conditions. CONCLUSION The above mentioned conditions breed a total content increase( of nearly 33%) of paeoniflorin and albiflorin,37. 60 mg / g,calculated at dry weight.