The objective of this study is to establish ISSR analysis system for Ping’ou hybrid hazelnut and analyze the genetic relationship for the main cultivars/varieties. 17 main cultivars/varieties, such as ‘Dawei’ and ‘Liaozhen No. 3’, are selected as materials. 7 ISSR primers are selected from 60 primers with high polymorphism, strong signals and clear bands. NTSYS pc 2.11F and Popgene 1.32 software are used to conduct the data analysis and plotting. The results show that: Totally 58 bands are obtained. The average number of bands is 8.3 per primer and the average percentage of polymorphic bands is 84.6%. All the samples are divided into 3 groups when the threshold value is 0.687. The first group, containing 14 cultivars/varieties, can be divided into four sub-groups, the second group contains 1 cultivar and the third group contains 2 varieties. The similarity coefficient of the samples varies from 0.448 to 0.879, with an average is 0.678. The effective number of alleles, Nei's gene diversity and Shannon's information index of the main cultivars/varieties are 1.6751、0.3701 and 0.5308, respectively. Ping’ou hybrid hazelnut is a high polymorphic population and the genetic relationships of the main cultivars are complex. The result of this study can offer reference for the relative researches in Corylus.