Abstract: Mitochondrial DNA (mtDNA) variation was investigated among 22 isofemale lines from different populations of Drosophila lini with 10 restriction endonucleases.The isofemale lines were collected originally from Taiwan,Guangdong in China and Yangon (Rangoon),Pyinoolwin (Maymyo)in Myanmar.Comparison of the cleavage sites allowed us to build a phylogenetic tree based on genetic distances of isofemale strains.The results show as follows:1)The mtDNA nucleomorph is the same in isofemale lines from Taiwan (TAW 3146.1) and Guangdong except DHS 307,315 and NKS 9243 from Guangdong.2)The difference is not found between MMY 307 and 326.3)The strains from Myanmar might have diverged.The genetic distance between TAW 3146.1 and RGN 3,RGN 206 is 0.00534 and 0.00553,between MMY 307,326 and RGN 3,RGN 206 is 0.00835 and 0.00868.4)The genetic distances between MMY 307,326 and TAW 3146.1 is very close (0.00274).As the same as between DHS 307,315,NKS 9243 and TAW 3146.1.