Based on genetic diversity analysis with ISSR-PCR, this study was to access the germplasm resources of Platycodon grandiflorum in northern Anhui province. Ten primers that could produce more distinct and repeatable bands were used for ISSR-PCR. Statistic analysis was conducted by POPGENE v. 1.32, Arlequin3.l, NTSYS-pc version 2.1. (1) Seventy-four polymorphic bands (76.29 %) out of a total of 97 were generated from 105 individuals in five populations. (2) Shannon index of diversity ranged from 0.307 to 0.260, diversity at species level was 0.3581, which means superior genetic diversity. (3) Genetic diversity across all the populations revealed by AMOVA indicated that 86.02 % occurred within populations. (4) The Fst value was 0.1398, indicating a intermediate genetic differentiation among populations. (5) Dendrogram relationship illustrated genetic distance was correlated with geographic distance. ISSR markers can be used for studying genetic diversity of P. grandiflorum. Degradation of populations doesn't happen in northern Anhui province, bank of germplasm preservation should be established for cultivation of excellent variety of P. grandiflorum.