Optimization Of Medium And Elicitors Treatment Research Of Salvia Miltiorrhiza Hairy Root

Salvia miltiorrhiza is a kind of perennial labiatae herbaceous plants, its root canactivate blood circulation, remove stasis pain, invigorate the circulation ofmenstruation and treat the cardiovascular disease, etc. It s mainly medicinalcomponent is tanshinone. In recent years, many studies have been reported aboutimproving the production of tanshinone through metabolic engineering strategies andtissue culture techniques; this provides a way to stability of the large-scale productionof tanshinone.This paper takes Salvia miltiorrhiza hairy root which has been imported keyenzyme genes SmGGPPS successfully as material. This paper attaches moreimportance to optimize the medium and finds optimum nutrition formula of medium,then chooses proper elicitors to improve the production of tanshinone.The mainresults of this paper are listed as followed:1. This paper took1/2MS medium as the basic medium and designedPlackett-Burman methods using Minitab software to filter significant factors thathad impacted on transgenic hairy root growth. The results that showed the descendingorder of significance were major elements, trace elements, vitamin, iron, inositol andglycine.2. Based on the results of Plackett-Burman methods, Box-Behnken methodswere designed to optimize the medium conditions of salvia miltiorrhiza hairy root.The results showed that: The transgenic strain required higher concentration of majorelements and vitamin obviously, but it needed lower concentration of the traceelements in medium. Further analysis showed that the medium composition was theconcentration of major elements corresponding to1.7311times, the concentration oftrace elements corresponding to0.5234times, the concentration of vitamincorresponding to2.0215times of original concentration.Compared with originalmedium, the hairy root biomass increased47.99%, total tanshinone productionincreased about45%in the hairy root.3. Finally, the bacteria and yeast extract were used to induce the hairy root in its exuberant growth period to further improve the production of tanshinone and to find akind of new and effective biological elicitors that can promote tanshinone synthesis.The results showed that: Yeast extract could improve the production of dihydrogentanshinone and cryptotanshinone obviously, the maximum multiples wererespectively3.38times and3.37times, but it made little effect on tanshinone I andtanshinone IIA. Pseudomonas aurantiacaon s supernatant fluid and cell homogenateshad certain inducing effect on tanshinone. Its induced effect on total tanshinones was1.49times, on dihydrogen tanshinone was1.37times, on cryptotanshinone was1.65times, on tanshinone I was1.45times, on tanshinone IIA was1.78times. But theresults of Bacillus subtilis and Bacillus licheniformison were not obvious.On the basis of previous studies, this experiment explores the nutritionalrequirement changes of transgenic hairy root growth and improves the salviamiltiorrhiza hairy root growth maximizely. Compared to the effect of yeast extract,this paper explores the effect of Pseudomonas aurantiacaon for the first time andmake up the blank for the transgenic hairy root, also lay some foundation for massindustrial production of salvia miltiorrhiza.