The Effect of The Combination of Precursor and Elicitors Enhance the Taxol Accumulation in Red Pine Cell Cultivation

Abstract

The material introduced into the callus culture was the in vitro cloned red pine stem and leaves. Callus obtained through culture from leaves and stems were included in the study on proliferation on agar and liquid media. The suspension obtained after 45 days of culture was determined for biomass, used in proliferation culture, and studied taxol accumulation. Media for taxus cell cloning was MS supplemented with 3 mg/l 2.4D, 3 mg/l NAA, 0.5 mg/l kinetin, 0.1 mg/l BA, 10% CW. Selections were carried out through 8 steps in the year of 2010 with interval cultivation time of 45 days/each step. It’s could improve the taxol accumulation via cell suspension cultures sourced from leaves 170.1 mg/gDW and stems 27.3 mg/gDW. Picloram was not effect on taxol accumulation. On the basic media MS supplemented with 3 mg/l 2.4D, 3 mg/l NAA, 0.5 mg/l kinetin, 0.1 mg/l BA, 10% CW supplemented with precursor of 15mg/l phenyl alanine (PA) effects on the percentage of FW/DW (fresh weight/dried weight) was 9.815 and the taxol accumulation was 0.778%. The effects of elicitors supplemented to media with 10 mg/l methyl jasmonate (MJ) having 7.183 FW/DW and 0.273% taxol, 100 mg/l salysilic acid (SA) having 10.12 FW/DW and 0.094% taxol, 50mg/l chitosan having 10.09 FW/DW and 0.119% taxol, 5mg/l oligo-chitosan having 9.090 FW/DW and 0.778% taxol. The effects of the combination of precursor and elicitors (15 mg/l phenyl alanine, 10 mg/l methyl jasmonate, 5 mg/l O-chitosan, 100 mg/l salysilic acid) enhance the taxol accumulation to 1.003% in comparison of separately as PA having 0.114% taxol, PA+Ochi having 0.542% taxol, PA+MJ+Ochi having 0.564%. Cell cloning from leaves (1.701%) had the taxol accumulation more than from stem (0.273%).