Effect Of Elicitor On The Growth And Cephalotaxus Alkaloids Accumulation Of Cephalotaxus Mannii Suspension Cells

Methyl jasmonic acid (MJ), salicylic acid (SA) and fungal endophyte culture supernatant (FE) were used as elicitors in the study. Adding different concentration of MJ(50,100,150?mol/L), SA(25,50,100?mol/L), and FE(2,4,6mL) at different time (10,15,20d) in the process of culture respectively. The elicitors were added by single addition and compound addition, the effects of the elicitors on the growth and cephalotaxus alkaloids accumulation of cephalotaxus mannii suspension cells were studied by testing cell growth rate, cell activity, browning of cultures, phenylalanine ammonia-lyase vitality (PAL), glucose6-phosphate dehydrogenase vitality (G6DPH), and the yield of cephalotaxus alkaloids. The results were as follows:In the respect of cell growth, cell average specific growth of cells treated with different concentrations of MJ and FE was decreased. Cell average specific growth was inversely proportional to the concentrations of MJ and FE. The study discovered that cell average specific growth was increased by adding50?mol/L SA, and the cell average specific growth of cells treated with50?mol/L SA was1.09times higher than the control. The compound addition of the elicitors indicated that the damage of cells caused by MJ and FE could be remitted through adding low concentration of SA. Cell average specific growth of cells treated with50?mol/L SA and100?mol/L MJ was14.28%higher than the cell average specific growth of cells treated with100?mol/L MJ. Cell average specific growth of cells treated with50?mol/L SA and4mL FE was9.52%higher than the cell average specific growth of cells treated with4mL FE.In the respect of PAL vitality and culture browning, the study tested PAL vitality in72hours after cells treated with100?mol/L MJ,50?mol/L SA and4mL FE, respectively, and the values of PAL vitality were compared with the control. The results showed that PAL vitality values of cells treated with MJ?SA?FE were increased, and the values of PAL vitality reached the maximum values at24h after cells treated with the elicitors. Culture browning values were increased rapidly in5days after cells treated with MJ?SA and FE respectively. The culture browning values were in direct proportion to the concentrations of the elicitors, and the values of culture browning was enhanced by adding elicitors.in combination.In the respect of G6DPH, the study tested G6DPH vitality in72hours after cells treated with100?mol/L MJ?50?mol/L SA?4mL FE, respectively, and the values of G6DPH vitality were compared with the control. The results showed that G6DPH vitality was motivated after adding FE, and G6DPH vitality reached the maximum value at48h after cells treated with FE. But the vitality of G6DPH treated with MJ and SA had no significant difference compared with the control. Therefore, G6DPH was not the action point of MJ and SA.In the respect of the yield of cephaloteaxus alkaloids, the optimum concentrations of MJ, SA, and FE were100?mol/L,50?mol/L, and4mL respectively. The best added time of MJ, SA and FE were15d,15d,10d respectively. Cells treated with two elicitors harvested more yield of cephalotaxus alkaloids compared with the cells treated with one elicitor. The results showed that cells treated with100?mol/L MJ and50?mol/L SA at the same time obtained the best yield of cephalotaxus alkaloids, and the yield of cephalotaxus alkaloids was4.18times higher than the control.