New nuclear primers for molecular studies of Epinephelidae fishes
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TECHNICAL NOTE
New nuclear primers for molecular studies of Epinephelidae fishes G. C. Silva-Oliveira • A. B. C. Silva • Y. Oliveira • Z. P. Nunes • R. A. Torres I. Sampaio • M. Vallinoto
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Received: 16 August 2012 / Accepted: 22 August 2012 / Published online: 1 September 2012 Ó Springer Science+Business Media B.V. 2012
Abstract The EPIC strategy was used in the present study in order to develop molecular markers for Epinephelus itajara. Twelve primers were designed for six genomic regions. The results of the study provide a number of appropriate new nuclear DNA markers for future genetic studies in several endangered/overexploited grouper species. The description of these markers will also expand the number of molecular tools available for the reconstruction of the life history of the different grouper species, in particular those in urgent need of effective protection measures. Keywords
Goliath grouper Intron Nuclear markers
The development of effective strategies for the protection and management of population units demands a holistic
G. C. Silva-Oliveira A. B. C. Silva Y. Oliveira Z. P. Nunes I. Sampaio M. Vallinoto (&) IECOS, Universidade Federal do Para´, Campus de Braganc¸a, Alameda Leandro Ribeiro s/n 68.600-000, Braganc¸a, Para´, Brazil e-mail: [email protected] G. C. Silva-Oliveira Universidade Federal do Para´, Campus do Marajo´-Breves, Av Anaja´s, s/n 68800-000, Aeroporto, Breves, Para´, Brazil R. A. Torres Laborato´rio de Genoˆmica Evolutiva e Ambiental, Departamento de Zoologia, Centro de Cieˆncias Biolo´gicas, Universidade Federal de Pernambuco, Recife, PE, Brazil M. Vallinoto CIBIO/UP, Centro de Investigac¸a˜o em Biodiversidade e Recursos Gene´ticos da Universidade do Porto, Campus Agra´rio de Vaira˜o, 4485-661 Vaira˜o, Portugal
approach that integrates data on the life-history strategy of the species with the ecological characteristics of the populations (Begg and Waldman 1999). In this context, the genome can offer important insights for the development of effective strategies in the conservation of endangered species by providing estimates of genetic variability (Lee et al. 2010; Guerier et al. 2012). The family Epinephelidae includes a number of commercially-important fish species that are found in subtropical and tropical regions throughout the World (Sadovy and Eklund 1999). Unregulated fishery exploitation, together with the biological characteristics of these species, contributes to the difficulties of conserving stocks (Sadovy and Eklund 1999; Frias-Torres 2006). Of the 20 epinephelids occurring in Brazil, two species, Epinephelus itajara (Goliath grouper) and Mycteroperca marginata (Dusky grouper), have the most preoccupying conservation status, being classified by IUCN, as critically endangered and endangered, respectively. The EPIC (exon-primer intron-crossing) strategy has been widely used to sequence introns, primarily because it uses conserved regions of exons to anchor the primers for the sequencing of the adjacent introns (Lee et al. 2010). In the present study,
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