Effect Of Elicitors On Accumulation Of Salvianolic Acid And Secondary Metabolism Enzyme Activities In Salvia Miltiorrhiza Bunge Callus.

In this paper, PAL, TAT, C4H, PPO, protective enzymes activities and salvianolic acid accumulation were studied by the application of the five elicitors (Casein acids hydrolysate, CAH), Co2+,(Fusarium solani, FS),(Armillaria mellea, AM) and (salicylic acid,SA) through callus culture using Salvia miltiorrhiza Bunge seedling callus as experimental materials.It provided the scientific basis for improving salvianolic acid content and regulating metabolizability. The results are as follow:1. The measure conditions of Secondary metabolism enzymes in Salvia miltiorrhiza Bunge salvianolic acid were studied. The proper pH,reaction temperature,reaction time and substrate concentration of PAL is 8.8, 37℃, 60 min and 20 mmol?L-1,respectively; C4H is 7.6, 30℃, 30 min and 25 mmol?L-1;TAT is 7.3, 37℃, 30 min and 17 mmol?L-1 ; PPO is 6.0, 30℃, 30 min and 10 mmol?L-1。2. Casein acids hydrolysate(CAH), Co2+, Fusarium solani(FS), Armillaria mellea(AM) and salicylic acid (SA) elicitors have notable effects on callus culture growth, pH and conductivity. The results as following: CAH, AM and FS can induce callus growth in low concentrations, but restrain in high concentrations; Co2+, SA restrain callus growth in all experimental concentrations, and more serious as concentration adds. The change of pH adds first,and then descends with increase in CAH concentrations; descends with increase in FS concentrations, and no change for AM; enhances with increase in Co2+, SA concentrations.The change of conductivity adds first,and then descends with increase in CAH, SA concentrations;adds increase in FS concentrations, and no change for AM, Co2+.3. The five elicitors have notable effects on caffeinic acid, salvianolic acid B. CAH, SA, Co2+ induce caffeinic acid, salvianolic acid B in low concentrations, and restrain in high concentrations; SA also induces salvianolic acid B into culture; FS induces in all experimental concentration; and AM restrain caffeninic acid ,but has no effect on salvianolic acid B.4. SA induces PAL, TAT, PPO activities, and the induced peak has relativity with SA concentration, the higher of SA concentration, the advanced of induced peak; SA restrains C4H activities. In a word, PAL, TAT are the key enzyme phenolpropanoid, but C4H is not.5. SA induces SOD, POD, CAT activities,but the induced time and intensity are different. The higher of SA concentration, the advanced of induced peak ; There SA concentration induce POD activities peak at the same time of 8 h; the effect of SA on CAT is reverse with SOD induced peak. All SA concentrations can induce dissolubility protein with no difference, and also induce dissolubility protein into culture.