Studying the genetic diversity and population structure of natural forest populations is essential for evaluating their ability to survive under future environmental changes and establishing conservation strategies. Pinus koraiensis is a conifer species with high ecological and economic value in Northeast China. However, its natural forests have been greatly reduced in recent years, mostly due to over exploitation and over utilization. Here, we evaluated the genetic diversity and population structure of seven populations of P. koraiensis located throughout its native distribution. A total of 204 samples were genotyped with nine polymorphic nuclear SSR (simple sequence repeat) markers. The results showed high genetic diversity in all populations, with an average expected heterozygosity of 0.610, and the northern-most populations (Dailin (DL) and Fenglin (FL)) showed slightly higher diversity than the other five populations. The level of genetic differentiation among populations was very low (F_ST = 0.020). Analysis of molecular variance (AMOVA) showed that only 2.35% of the genetic variation existed among populations. Moreover, STRUCTURE analysis clearly separated the seven populations into two clusters. Populations DL and FL from the Xiaoxinganling Mountains comprised cluster I, while cluster II included the five populations from the Changbai Mountains and adjacent highlands. Our research on the genetic diversity and population structure of P. koraiensis in natural forests of China can provide a basis for the implementation of programs for the conservation and utilization of P. koraiensis genetic resources in the future.