DITERPENOID PRODUCTION IN HAIRY ROOT CULTURES OF SALVIA-MILTIORRHIZA
DITERPENOID PRODUCTION IN HAIRY ROOT CULTURES OF SALVIA-MILTIORRHIZA
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DOI：
10.1016/s0031-9422(00)95156-2
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发表时间：
1993-02-01
期刊：
影响因子：
3.8
通讯作者：
ALFERMANN, AW
中科院分区：
文献类型：
--
作者：
HU, ZB;ALFERMANN, AW
用发根农杆菌菌株LBA9402、ATCC 15834、TR105、R1601和A41027侵染无菌植株，建立了丹参转化根培养体系。这些菌株在该植物上诱导毛状根的能力不同。此外，还观察到乙酰丁香酮对农杆菌介导的丹参毛状根分化有促进作用。所有转化的根培养物都进行了TLC检测，农碱和甘露碱均呈阳性。建立了同时测定丹参酮I、丹参酮II(A)、丹参酮II(B)、丹参酮V、二氢丹参酮I、隐丹参酮、丹参酮VI和无色二萜铁锈醇的高效液相色谱方法。这7种主要的丹参酮和铁锈醇不仅存在于根部，也存在于液体培养基中。显然，毛状根将次生代谢物释放到培养基中。毛状根培养的主要产物是丹参酮类化合物，丹参酮是丹参酮的活性成分。研究了培养条件对毛状根生长和二萜产量的影响。当改变介质中的硝酸铵水平时，效果最明显。
Transformed root cultures of Salvia miltiorrhiza have been established by infecting pieces of sterile grown plants with Agrobacterium rhizogenes strains LBA 9402, ATCC 15834, TR 105, R 1601 and A 4 1027. These strains showed different abilities to induce hairy roots on this plant. A promoting effect of acetosyringone on the Agrobacterium-mediated hairy root initiation in Salvia miltiorrhiza was also observed. All transformed root cultures were assayed for opines by TLC, and were positive for agropine and mannopine. A sensitive quantitative HPLC procedure was developed for the simultaneous quantification of tanshinone I, tanshinone II(A), tanshinone II(B), tanshinone V, dihydrotanshinone I, cryptotanshinone, tanshinone VI, and the colourless diterpene ferruginol. These seven major tanshinones and ferruginol were found not only in the roots, but also in the liquid medium. Apparently, the hairy roots released secondary metabolites into the culture medium. The main products in the hairy root cultures were the tanshinones which are the active principles of this plant species. The effects of culture conditions on hairy root growth and the diterpene production have also been studied. The most apparent effect was seen when ammonium nitrate levels of the medium were modified.