欧洲缬草不定根悬浮培养及其有效成分的测定

为高效生产欧洲缬草有效成分,通过分析培养基pH值、碳源、糖浓度、盐浓度、总氮(N)浓度、总磷(P)浓度等因素对其不定根悬浮培养生长的影响,确定欧洲缬草不定根悬浮培养的适宜条件,并测定在该培养条件下不定根的生长状况,同时比较正常不定根、褐化不定根、无菌苗根中缬草三酯、乙酰缬草三酯和缬草烯酸3种有效成分的含量。结果表明,欧洲缬草不定根的最佳培养条件为2倍大量元素浓度、48.46 mmol·L^-1总N浓度、1.5 mmol·L^-1总P浓度的RCM培养基,添加40 g·L^-1蔗糖、0.5 mg·L^-1 萘乙酸(NAA),pH值5.7,温度25±2℃,转速100 r·min^-1暗培养,28 d后收获。最佳培养条件下的欧洲缬草正常不定根中缬草三酯、乙酰缬草三酯、缬草烯酸的含量均最高,分别为7.222、0.355、4.016 mg·g^-1,褐化不定根中成分含量显著降低,无菌苗根中成分含量最低。本研究结果可为欧洲缬草不定根及其有效成分的规模化生产提供参考。

Suspension Culture of Adventitious Root and Determination of Active Ingredients of Valeriana officinalis

The adventitious roots of Valeriana officinalis could synthesize secondary metabolites stably, and tissue culture techniques have quick reproduce and large reproductive coefficient. In order to product efficiently the active ingredients of Valeriana officinalis. The effects of pH value, carbon source type, sugar concentration, salt concentration, total N concentration and total P concentration on the suspension culture have been analyzed: The optimal suspension culture condition was gained for adventitious root. The growth curve has been constructed. Meanwhile the content of valepotriate, acevalepotriate, and valerenic acid in normal adventitious roots were contrasted, browning adventitious roots and aseptic seedling roots. The results showed that the ideal suspension culture condition of adventitious roots was twice major element concentration, 48.46 mmol·L^-1 total N concentration, 1.5 mmol·L^-1 total P concentration in RCM medium, and 40 g·L^-1 sucrose, 0.5 mg·L^-1 NAA, pH 5.7, cultured at 25±2℃, 100 r·min^-1 in dark then harvested the adventitious roots after 28 days inoculation. It was also found that in this condition the content of valepotriate, acevalepotriater, and valerenic acid were respectively 7.222 mg·g^-1, 0.355 mg·g^-1, and 4.016 mg·g^-1, the content of the components in browning root significantly decreased. The content of the components in the aseptic seedling roots was the least. It also provided reference for the adventitious roots induction and the active ingredients production of V. officinalis.