Abstract: Seeds collected from eight populations of Chir pine (Pinus roxburghii SARG.) from the natural distribution range of the species in Himachal Himalayas in India were analysed isozymatically at 11 enzyme systems. For the enzyme systems studied, 25 gene loci were identified out of which 18 were polymorphic. The observed mean values for genetic variation were slightly lower than mean values reported for Pinus species (number of alleles: 1.65 compared to 2.36; effective number of alleles: 1.13 compared to 1.26; observed heterozygosity: 0.153 compared to 0.179). A small differentiation among populations and large variation within populations were reflected by small value of GST (0.04): Considering the different genetic parameters three populations seem favourable for gene conservation measures.