Effects Of Salicylica Acid Induced Salvia Miltirriaza Supension Cells On Secondary Metablism, Cytoalsmc Acidifcation And Active Oxygen

In this paper, Salvia leaves callus for cell suspension cultures was using as the research material;Salicylic acid was using as the elicitor;nitrendipine, verapamil and EGTA were using as calcium inhibitors. After adding Salicylic acid elicitor and calcium inhibitors , detectd respectively the content of Salvia phenolic secondary metabolites in cell culture , the pH changes of cell culture medium in inducing , the content of hydrogen peroxide and the activation of enzyme related (SOD,POD,CAT,PPO). The sdudy research the effects of alicylic acid induced Salvia miltiorrhiza suspension cells culture on secondary metabolites biosynthesis,cytoplasmic acidification and oxidative burst of Salvia miltiorrhiza cell .And research discuss the mechanism of action on salicylic acid induced the above change, in order to provide the basis research for using cell culture to produce secondary metabolites and preliminary understanding salicylic acid induced cellular signal transduction mechanisms. The results are as follow:1.A lower concentration (3.125²5 mg/L) of salicylic acid can be used as a abiotic elicitor adjusting the secondary metabolite of Salvia miltiorrhiza, inducing Salvia miltiorrhiza cell water-soluble phenols secondary metabolites biosynthesis. The suitable concentration of salicylic acid induced salvia phenolic secondary metabolites is between 6.25²5 mg/L, the value of salvia cultured cells biosynthesis salvianolic acid B and caffeic acid was up to 0.62% and 0.048% . It was the 1.82 times and 2.67 times , compared to the control respectively.2.Using the optimal concentration of salicylic acid (6.25mg/L) induced Salvia miltiorrhiza cell culture , the biosynthesis accumulation curves of the salvianolic acid B and caffeic acid was both the same . Started after induction of 2 7 d, the salvianolic acid B and caffeic acid is increasing rapidly. At the induction of the first 6 d, the content of salvianolic acid B arrived at maximum, the content of caffeic acid in the first 7 d arrived at maximum. Therefore, the better time of the induction of the above two secondary metabolites is 6 7 d.3.Salicylic acid can induce Salvia miltiorrhiza cell culture medium H⁺ influx, leading to acidification of the cytoplasm culture of Salvia miltiorrhiza. The extent of cytoplasmic acidification and the time were relatead to the concentration of salicylic acid elicitor in a concentration dependent manner. The higher the concentration of salicylic acid, the eariler salvia miltiorrhiza cell suspension culture medium pH changes in the maximum occured. Calcium inhibitors can reduce the rate of salicylic acid-induced pH changes. Salicylic acid-induced cytoplasmic acidification of Salvia miltiorrhiza cell depends on the calcium burst inside and outside the Salvia miltiorrhiza cell culture , can be reduced by calcium inhibitor.4.Salicylic acid can induce Salvia cultured cells oxidative burst and increase the content of H₂O₂. After 2 h of the induction, H₂O₂ content in the cells achieve 1.9 times to the control. After 4 h of the induction, H₂O₂ content in the cells return to control level. Calcium ion inhibitor nitrendipine, verapamil and EGTA inhibited cell calcium levels effectively, thereby inhibited salicylic acid-induced Salvia cells in the generation of H₂O₂. Salicylic acid-induced Salvia cells H₂O₂ dependence the extra and intracellular Ca²⁺.5.Salicylic acid induced the increased activity of Salvia cells PPO and SOD. Calcium inhibitors inhibited results that salicylic acid stimulated activity of SOD and PPO. The inhibiting of nitrendipine and verapamil on the SOD activity after the induction of salicylic acid had effects of concentration, while the EGTA concentration had little to do with activity change of SOD and PPO. Cell calcium involved in the regulation of elicitor-induced miltiorrhiza cell culture activity of SOD and PPO.6.Salvia cultured cells activity of POD and CAT induced by Salicylic acid is down sharply. Calcium ion inhibitors can suppress decreasing of Salvia cell suspension POD and CAT activity salicylic acid-induced. The inhibiting of nitrendipine and verapamil on the POD and CAT activity after the induction of salicylic acid had effects of concentration, while the EGTA concentration had little to do with activity change of POD and CAT. Cell calcium involved in the regulation of elicitor-induced miltiorrhiza cell culture activity of POD and CAT.