Transformation of Hevea Suspension Cells by Electroporation
The uniform design coupled with Partial Least-squares(PLS)regression synthetical was utilized to evaluate the comprehensive test indices as transformation efficiency(Y1)and cell survival rate(Y2)by analyzing Hevea Rubber Hybrid B1(H.brasiliensis×H.nitida)suspension cells.Regression modeling of two test indices were built to optimize the maximum value of the dependent variable.The results showed that under the electric shock conditions of 10 μF,2 kn,1 300 V/cm,a final concentration of 90 μg/mL linear plasmid CaMV35S-EGFP-NOS in 0.4 mol/L sucrose electroporation solution and one minute of 600 mbar vacuum-pumping,logarithmic phase suspended cells,cultured 3 d,could reach an instant expression efficiency and survival rate in 0.648 1% and 56.984 7%,respectively.This study established a system that was applicable to a stable electric shock of Hevea rubber suspension cells and provided an effective method for the research of Hevea rubber suspension cells transgene by optimizing some correlative factors during electroporation.