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作者前期研究成果显示密旋链霉菌Act12可以上调丹参酮生物合成途径上的关键酶基因的表达大幅提高丹参毛状根中丹参酮含量。该实验则进一步研究了活性氧在Act12诱导丹参毛状根中丹参酮积累的作用。在继代培养21 d的丹参毛状根中添加Act12不同的诱导子及诱导子组合,分别测定不同收获期毛状根的生物量,毛状根中活性氧积累量,丹参酮类成分的积累量和关键酶基因表达量。Act12诱导后丹参毛状根中活性氧含量上升,HMGR和DXR基因表达上调,最高分别达到对照的32.4,4.8倍,毛状根中丹参酮积累增加,最高达到对照的10.2倍;加入活性氧抑制剂CAT和SOD后,丹参毛状根中的活性氧含量较Act12处理显著下降,HMGR和DXR基因表达也明显下降,毛状根中丹参酮含量较Act12处理下降了74.6%。活性氧介导了Act12诱导丹参酮积累,Act12很可能是通过ROS信号通路激活了MVA和MEP途径,从而提高了丹参酮在毛状根中的含量。
The author’s previous research results show that Act12 can up-regulate the expression of key enzyme genes in the pathway of tanshinone biosynthesis, and significantly increase the content of tanshinone in the hairy roots of Salvia miltiorrhiza. In this experiment, the effect of reactive oxygen species on tanshinone accumulation in Salvia miltiorrhiza hairy roots was further studied. Act12 different inductors and elicitor combinations were added into the hairy roots of Salvia miltiorrhiza for 21 days. The biomass of hairy roots, the accumulation of active oxygen in hairy roots and the accumulation of tanshinones And key enzyme gene expression levels. Act12 induced reactive oxygen species in hairy root of Salvia miltiorrhiza increased, the expression of HMGR and DXR gene increased up to 32.4 and 4.8 times of the control respectively, and the accumulation of tanshinone in hairy root increased up to 10.2 times that of the control; addition of reactive oxygen species inhibitor Compared with Act12 treatment, the content of reactive oxygen species in hairy root of Salvia miltiorrhiza Bunge significantly decreased, while the expression of HMGR and DXR gene also decreased obviously. The content of tanshinone in hairy root decreased by 74.6% compared with Act12 treatment. Act12 mediates Act12-induced tanshinone accumulation. Act12 probably activates the MVA and MEP pathways through the ROS signaling pathway, thereby increasing the content of tanshinone in hairy roots.