The genetic variation in populations of Anatolian black pine (Pinus nigra Am. subsp. pallasiana (L.) Holmboe.), one of the species covering large areas in Turkey, was investigated. Open pollinated seeds were collected from 13 populations in a natural distribution range. Six characters of seeds (length, width, ratio of length to width, weight/1000 seeds) and seedling characters (cotyledon number and hypocotyls height) and two enzyme systems viz. leucine aminopeptidase (LAP) and glutamate oxaloacetate transaminase, (GOT) were investigated. Significant differences were detected among the populations for the morphological characters. In addition, isozyme patterns of two enzyme systems revealed that LAP has two loci (one with 2 alleles and the other with 3), while GOT has three loci (two with 3 alleles and the third one with 2 alleles). Polymorphic loci were 74% on the average. The mean number of alleles per loci was 1.94 and expected heterozygosity was 19%. The mean total genetic diversity was calculated as 0.203; the mean gene diversity within populations was determined as 0.188, and the average between subpopulations diversity was 0.016. The relative magnitude of genetic differentiation among subpopulations was measured as 0.074 indicating that only 74% of the total genetic diversity was there between populations. Average genetic distance was 0.093 according to Gregodius. Nei's genetic distance was 0.022.