Objective:To establish the inter simple squence repeat(ISSR)-PCR reaction system of Tupistra chinensis. Method: The total genome DNA of T. chinensis. was used as the material. The concentration of Mg2+

dNTP

primer and Taq DNA polymerase

which greatly influence ISSR-PCR reaction

were optimized. Result: The optimized reaction conditions was established as follows:in a volume of 10 μL containing 1×buffer

2 U Taq DNA polymerase

2 mmol·L-1 MgCl2

0.2 mmol·L-1 dNTP

0.75 μL primer. Conclusion: Sixteen primers with stable amplification and rich polymorphism were screened from 80 (inter simple sequence repeat)ISSR primers. The optimized annealing temperature for each primer in ISSR-PCR reaction was proposed. This study will be helpful for research on the genetic diversity analysis and germplasm identification of T. chinensis.