为筛选悬浮培养福建金线莲丛生芽中酚和黄酮积累的适宜培养基,对MS基本培养基浓度、苄基腺嘌呤(BA)、激动素(KT)及萘乙酸(NAA)浓度的4因素8水平及蔗糖、磷和氮浓度3因素7水平利用均匀设计法,研究了2组均匀设计的不同组合处理对福建金线莲丛生芽中酚和黄酮积累的影响。同时,对培养基中NO3^-和NH4^+浓度比的影响也进行了研究。结果表明:酚和黄酮的最高含量和生产量出现在3/4 MS+BA 2.0mg/L+KT 0.2mg/L+NAA 0.5mg/L组合和蔗糖35g/L+磷0.93mM+氮45mM组合。培养基中总氮浓度为45mM的条件下,对NO3^-和NH4^+浓度的影响进行了调查,结果发现当NO3^-和NH4^+浓度分别为30和15mM时,最利于酚和黄酮积累,总酚和总黄酮生产量分别达到112.7和87.9mg/L。因此,在福建金线莲丛生芽悬浮培养时,将3/4MS培养基中的总磷浓度调节为0.93mM,NO3^-和NH4^+浓度分别调节为30和15mM,加入BA 2.0mg/L,KT 0.2mg/L,NAA 0.5mg/L和蔗糖35g/L时可获得较多的酚和黄酮。
In order to screening out a suitable culture medium for phenolic and flavonoid accumulation of adventitious buds during suspension culture,four factors and 8 levels with MS medium strength,concentrations of benzyladenine(BA),kinetin(KT),andα-naphthylacetic acid(NAA)and the 3 factors and 7 levels with concentrations of sucrose,total phosphorus,and total nitrogen were designed by the uniform design method,and their effects on phenolic and flavonoid accumulation were investigated.The results showed that among the combinations of MS medium strength,BA,KT,and NAA,phenolic and flavonoid contents and productivities were favorable at the combination of 3/4MS+2.0mg/L BA+0.2mg/L KT+0.5mg/L NAA;among the combinations of sucrose,phosphorus,and nitrogen,the combination of 35g/L sucrose+0.93 mM phosphorus+45mM nitrogen was optimal.The ratio of NO3~-/NH4~+ also had an influence,the maximum phenolic and flavonoid accumulation were found at the NO3^-/NH4^+ ratio of 30/15 mM,where the productivities of total phenolics and flavonoids reached 112.7 mg/L and 87.9mg/L,respectively.Therefore,the suspension culture of adventitious buds for phenolic and flavonoid production,the suitable culture medium was 3/4 MS culture containing 0.93 mM phosphorus,30 mM NO3^- and 15 mM NH4~+supplemented with 2.0mg/L BA,0.2mg/L KT,0.5mg/L NAA,and 35g/L sucrose.