Abstract: 【Objective】 This study aimed to identify cyst nematode resistance genes in potato germplasm resources through molecular marker， so as to enrich the gene resources for potato cyst nematode resistance and provide important resistance sources for breeding potato resistant to cyst nematode. 【Method】 320 potato germplasm resources introduced from the International Potato Center were used as materials. The amplification efficiency and specificity of tightly linked molecular marker primers， GBSS1-3， N146， N195， Gro1-4-1， and GPA2-2 for the granule-bound starch synthase 1 as an internal reference gene in potato （GBSS1）， Globodera rostochiensis resistance genes （H1 and Gro1-4）， and Globodera pallida resistance genes （Gpa2） were validated through conventional PCR. As the multiplex PCR system was optimized， three potential cyst nematode resistance genes， H1， Gro1-4， and Gpa2 in 320 potato germplasm resources were screened through high-throughput rapid detection， and the composition of resistance genes of 320 germplasm resources was analyzed by Venn diagram. 【Result】 A multiplex PCR system with five molecular markers was developed to rapidly detect resistance genes of potato cyst nematode. A pattern map was constructed for simultaneous detection of three resistance genes （H1， Gro1-4， and Gpa2） against cyst nematodes based on multiplex PCR. The H1 gene was identified in 38 materials using molecular markers N146 and N195 at a ratio of 11.88%. No Gro1-4 gene was detected using the Gro1-4-1 marker. 197 materials containing the Gpa2 gene were identified by the Gpa2-2 marker， with a ratio of 61.56%. H1， Gro1-4， and Gpa2 genes were not detected in 108 materials that accounted for 33.75%. The analysis of the composition of resistance genes revealed that 23 resources contained both H1 and Gpa2 resistance genes， with a ratio of 7.19%. No material contai-ning the three resistance genes was identified. 【Conclusion】 A pattern map for the simultaneous detection of potato cyst nematode resistance genes （H1， Gro1-4， and Gpa2） by multiplex PCR is established. Twenty-three materials contain both H1 and Gpa2 genes， and these dual-resistant potato germplasm resources combining H1 and Gpa2 genes could be utilized as excellent resistance sources for breeding potato resistant to cyst nematode.