Abstract: Genetic diversity of 25 accessions of the medicinal climber Tinospora cordifolia was measured by isozymes from Gujarat. The germplasm was reared in a field plot under identical conditions and mature stems were used for the presented study. Analysis by using battery of ten isozymes revealed the presence of 16 gene loci and 33 alleles in 25 accessions. The percentage of polymorphic loci (P) was 45.0% and mean observed number of alleles per locus (A) was 1.57. The average observed heterozygosity (Ho) and expected heterozygosity (He) were 0.443 and 0.270, respectively shows high levels of genetic variation among different accessions. The UPGMA dendrogram clearly depict the spectra of genetic diversity among various accessions. The clustering of accession TC-1 (Kheda) and TC-2 (Songhad) appeared at the top of the dendrogram which are genetically rich. These accessions should be conserved for future breeding programme.