Keywords: 
• Pogostemon cablin /
• ISSR-PCR /
• single factor tests /
• orthogonal design

Abstract: In order to establish the optimized ISSR-PCR experiment system for Pogostemon cablin,the single factor experiments were used to detect the suitable concentration ranges of the different influential factors,and then the orthogonal design was performed to optimize 5 factors（Mg²⁺,dNTPs,primer,Taq polymerase,DNA template） at 4 levels,which affect ISSR-PCR amplification system of Pogostemon cablin.The results indicated that a suitable ISSR-PCR reaction system established: 10×buffer 2.5 μL,40 ng DNA template,2.5 mmol·L^-1 MgCl₂,0.3 μmol·L^-1 primers,1.5 U Taq polymerase,150 μmol·L^-1 dNTPs were contained in 25 μL reaction solution.The optimized amplification program was that predenaturing at 94 ℃ for 5 min,then denaturing at 94 ℃ for 45 s,primer annealing at 52.7 ℃ for 45 s,extension at 72 ℃ for 90 s,for 40 cycles,at last extension at 72 ℃ for 7 min,and the products were stored at 4 ℃.