以贯叶连翘种子为材料,对贯叶连翘组培苗进行生根培养,筛选和优化生根培养基,进一步在生物反应器内诱导培养不定根,并研究了蔗糖浓度、铵态氮/硝态氮配比以及MS培养基浓度对不定根中金丝桃素含量积累的影响。结果表明:贯叶连翘在3/4MS+IBA2.5mg/L+蔗糖30g/L+琼脂8g/L(pH5.8)培养基上的生根效果最佳;40g/L的蔗糖有利于金丝桃素的积累,此时不定根中金丝桃素的含量最高、达0.94mg/g;当铵态氮/硝态氮配比为30/30时,金丝桃素的含量和生产量达到最大,分别为0.98mg/g和12.41mg/L;不定根在不同浓度范围的MS培养基中均可生长,金丝桃素在2MS培养基上的生产量最大、为15.70mg/L。
In this study, the seeds of Hypericum perforatum were used as the materials. Tissue culture seedlings rooting culture experiment was carried out to screen and optimize the rooting culture medium. Then the adventitious roots were induced and cultured in bioreactor. The effects of sucrose concentration, NH4+/NO3- ratio and MS strength on hypericin content of adventitious roots in bioreactor culture were investigated. The results showed that tissue culture seedlings rooted best with IBA 2.5 mg/L, sucrose 30 g/L and agar 8 g/L (pH 5.8) in 3/4MS medium. 40 g/L of sucrose concentration was conducive to the accumulation of hypericin, the hypericin content was highest (0.94 mg/g). When NH4+/NO3- ratio was 30/30, hypericin content and productivity reached the maximum, the values were 0.98 mg/g and 12.41 mg/L respectively. Adventitious roots could grew in different MS strength medium, hypericin produced the largest amount (15.70 mg/L) in 2MS strength.