Studies On Components Accumulation And Relevant Key Enzyme Genes Expression Of Gentiana Macrophylla Pall. From Shannxi

Genuine medicinal materials has reliable curative effect and stable active components,with a long clinical screening.Studying the quality formation of Genuine medicinal materials can provide a theoretical basis for optimizing the quality and improving the curative effect of material medica.Active components is basic function substance of traditional Chinese medicine and directly affect the curative effect of medicine.The synthesis and accumulation of active components is the key to the quality formation of genuine medicinal materials.In this paper,the quality formation of genuine Gentiana Radix from Shaanxi is studied taking Gentiana macrophylla Pall.as research object.The contents of five iridoids,main active constituents of Gentiana Radix and expression of three relevant key enzyme genes are measured under different conditions.And the effects of different dehydration methods on Gentiana Macrophylla Radix also was studied.The results are as follows:From end of June to beginning of September,the content of active components in different age Gentiana macrophylla Pall.is in the vicinity of higher number,the content of I₁?loganic acid?in the aerial part is high,and the relative expression of G10H and SLS are also maintain at a higher level,while the relative expression of DXR is at a lower level;between the end of April and mid-October,contents of I₂?6'-O-?-glucosyl-gentiopicroside??I₃?swertiamarin??I₄?gentiopicroside?and total content of the five iridoids in underground part are significantly higher than aerial part,all three genes express in leaf,stem,and root,the expression abundance is the highest in leaf and the lowest in stem;there are significant positive correlations of the contents changes between different age;the change of four years old Gentiana macrophylla Pall.is more stable than two or three years old,changes of content have some relationship to the fluctuation of relative gene expression.Gentiana macrophylla Pall.has different response under different elicitors such as Me JA,SA,Ag₂SO₄,and Alternaria Nees solution,the response mechanisms of root,stem,and leaf to the same inducer are different,but there is a positive correlation between the expression changes of the three genes to the same inducer,and between the content changes of five active components there is also a positive correlation,too;when the content changes significantly or very significantly,it is accompanied by significant expression fluctuations of two or more genes among DXR,G10H,and SLS,or extremely significant by a single gene expression fluctuations.After 30 days of drought stress,the response of the stem of Gentiana macrophylla Pall.is lower than that of root and leaf.The contents of I₁,I₂,I₅?sweroside?in root,and I₂,I₄in stem,and I₃,I₄,I₅in leaf are significantly lower than the control group,spatial relevant synthase genes are significantly down-regulated in different parts;after 30 days of salt stress,the contents of I₁,I₂,I₅in root are significantly decrease,the content of I₄increases significantly,the content of I₁in stem increases significantly,and the content of I₄in leaf increases significantly with the increase of salt content,spatial relevant synthetase genes are significantly or extremely up-regulated in different part.The results show that compared with the medicinal materials dried without diaphoresis treatment,contents of I₁,I₄,and total content of the five constituents in dehydrated medicinal materials which has sweated,are very significantly lower,but content of I₂is significantly higher than crude drugs without diaphoresis,the contents of I₃and I₅have no statistical difference between sweating and no-sweating group;among these three ways of dehydrating:oven dry,dry in the sun and dry in shade,dry in shade is the best way for the preservation of I₄and total content of the five iridoid glycosides,while oven dry can be propitious to retain I₁,relatively,dry in the sun is adverse for main active constituents in Gentiana Macrophylla Radix;the antioxidative and antibacterial activity of the sweat treated herbs is higher than that of the non-sweating treatment,the antioxidative activities of the oven dried herbs are the highest and the ones which dried in the sun are the lowest.