Use of PCR-RFLP Analysis of mtDNA Cytochrome-b Gene to Determine Genetic Differences in Capoeta spp.

Abstract

Genetic differences between Capoeta capoeta capoeta, Capoeta capoeta umbla, and Capoeta tinca were determined using PCR-RFLP of mtDNA cytochrome-b (Cyt-b) by amplifying approxi- mately 400-500 bp of this region from each of the three subspecies. The restriction enzymes SpeI and HinfI did not indicate genetic differences but AluI and HpaII did. Thus, PCR-RFLP of the mtDNA was used to distinguish between closely related subspecies without having to ana- lyze the entire DNA sequence of specimens. Use of this technique demonstrated that the Cyt-b regions of the three subspecies have different base sequences. The similarity between C. c. capoeta and C. c. umbla is 71.4% while C. tinca is more distant (50% for C. c. umbla and 33.3% for C. c. capoeta). The Cyt-b similarity is consistent with morphological and taxonomical similar- ities. PCR-RFLP can serve as a tool for genetically identifying subspecies of fish in nature and in aquaculture.