Development of RAPD‐SCAR and RAPD‐generated PCR‐RFLP markers for identification of four Anguilla eel species

Abstract

Discriminating between eel species of the genus Anguilla using morphological characteristics can be problematic, particularly in the glass eel and elver stages. In this study, sequence‐characterized amplified region (SCAR) and polymerase chain reaction (PCR)‐restriction fragment length polymorphism (RFLP) markers were developed for the identification of Anguilla japonica, Anguilla bicolor bicolor, Anguilla rostrata, and Anguilla anguilla. Random amplified polymorphic DNA (RAPD) fragments from A. japonica (362 bp), A. bicolor bicolor (375 bp), A. rostrata (375 bp), and A. anguilla (375 bp) were isolated, sequenced, and converted to SCAR markers. The principal difference between the SCARs of A. japonica and the three other species is the absence of a 13 bp deletion in the A. japonica SCAR. Specific PCR primers amplified a 290 bp fragment for A. japonica and 303 bp fragments for A. bicolor bicolor, A. rostrata, and A. anguilla. Restriction enzyme digestion with TaqI, MaeI, and Tru9I yielded PCR‐RFLP patterns with differences that, when analyzed together, are sufficient for distinguishing each of the four eel species. In addition, RAPD fragments for A. japonica (577 bp), A. bicolor bicolor (540 bp), A. rostrata (540 bp), and A. anguilla (509 bp) were also isolated and sequenced. The A. japonica, A. bicolor bicolor, A. rostrata, and A. anguilla PCR products contain ten, nine, nine, and eight tandem repeats, respectively, of a 37 bp sequence. These results suggest that SCAR and PCR‐RFLP markers and repeat numbers for specific loci will be useful for the identification of these four Anguilla eel species.

Key words:

• Anguilla
• SCAR
• PCR‐RFLP
• species identification

Notes

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