本研究采用L16(45)正交试验设计,对油用牡丹ISSR反应体系中的5个主要因素进行了优化,确立了ISSR-PCR的最佳反应体系:反应体系总体积20μL,含0.50 U Taq聚合酶,20 ng模板DNA,3.00 mmol/L Mg~(2+),0.40 mmol/L d NTPs,1.00μmol/L引物及10×PCR Buffer。基于ISSR-PCR最佳反应体系,我们共筛选出了12条多态性丰富、条带清晰稳定的引物,为油用牡丹遗传连锁图谱的构建及分子育种奠定基础。 In this study, L16 (45) orthogonal design was used to optimize the five main factors in ISSR reaction system of peony. The optimal reaction system for ISSR-PCR was established. The total volume of the reaction system was 20 μL, containing 0.50 U Taq Polymerase, 20 ng template DNA, 3.00 mmol / L Mg 2+, 0.40 mmol / L d NTPs, 1.00 μmol / L primer and 10 × PCR Buffer. Based on the ISSR-PCR optimal reaction system, we screened 12 primers with abundant polymorphism and clear and stable bands, which laid the foundation for the genetic linkage map construction and molecular breeding of peony.