DNA Fingnerprinting of Eight Cyprinid Fish Species of Iraqi Inland Waters Using RAPD-PCR Technique

Abstract

One hundred and twenty specimens of eight cyprinid fish species included <i>Barbus xanthopterus, B. kersin, B. barbulus, B. grypus, B. sharpeyi, B. luteus, Aspius vorax </i>and<i> Cyprinus carpio</i> were collected from Shatt Al-Arab River in governorate of Basrah Southern of Iraq<i>. </i>The DNA fingerprints were identified using RAPD-PCR technique with seven decamere primers. A total of 223 anonymous scorable bands obtained by RAPD-PCR technique across eight cyprinid species genomes. The average of bands per primer was 32 bands, the size of bands estimated using the linear equation of standard curve plot of standard 100 bp ladder. The RAPD markers were used to obtain the similarity indices that were in high value among<i> Barbus</i> species since it ranged from 84.4% between <i>B. xanthopterus </i>and <i>B. kersin</i> to 52% between <i>B. barbulus</i> and <i>B. sharpeyi</i>. While the <i>B. grypus </i>appeared in high<i> </i>similarity indices value (86.9%) with luciscin<i> A. vorax. </i>While <i>C. carpio</i> exhibit high diversity indices value with the <i>Barbus</i> spp.. Therefore the phylogenetic relationships among the eight studied cyprinid species were clearly resolved in dendrograms, they clustered into three branches: the first one included <i>B. xa</i><i>n</i><i>thopterus, B. kersin, B, barbulus,</i> while the second included <i>B. sharpeyi and B. luteus</i> and the third branch contained <i>B. grypus</i> and <i>A. vorax</i>. Nevertheless, when <i>A. vorax</i> data discarded, the <i>B. grypus</i> return to cluster with <i>B. sharpeyi</i> and <i>B. luteus</i>. Whereas <i>C. carpio</i> functioned as outgroup in both cases. The conclusion of the present study was: molecular patterns showed by RAPD bands represent specific DNA fingerprints and apparently are suitable tool to distinguish among cyprinids. The results clearly revealed that the eight cyprinid taxa are distinctive species. The studied <i>Barbus</i> species are divided into two groups, the first one included <i>B. xanthopterus</i>, <i>B. kersin</i> and <i>B. barbulus</i> which have European<i> Luciobarbus</i> affinities and the second contained <i>B. grypus</i>, <i>B. sharpeyi</i> and <i>B. luteus</i> which they diverged of <i>Luciobarbus</i> but still related to <i>Barbus</i>. So, the <i>Barbus</i> in Iraq is polyphyletic group.