Abstract: In this study, orthogonal experiments were designed to optimize the key parameters (dNTPs、Taq polymerase、primer、template DNA) of SRAP-PCR amplification system in loofah（Luffa cylindrica (L.) M. Roem）. The results showed that the optimized reaction system included: DNA template 50ng, dNTPs 0.3mmol/L, primer 0.30μmol/L and Taq polymerase 1U in the 10ul total volume. This system is fit for the genome amplification of loofah and can be well used for the molecular biological research in loofah.