In an attempt to identify a simple, reproducible, inexpensive and non-toxic method for obtaining high quality and quantity genomic DNA from fi sh fi n and larvae samples, modifi ed salt extraction protocol and modifi ed phenol-chloroform extraction protocol were compared. The samples obtained from different fi sh species (Brycon orbignyanus, Piaractus mesopotamicus, Oreochromis niloticus, Leporinus elongatus and Prochilodus lineatus) using salt extraction showed good DNA quality and quantity (DNA/RNA relationship 1.8-2.2). These DNA samples were successfully used in the amplifi cation of RAPD (Random Amplifi ed Polymorphic DNA) and microsatellite molecular markers, demonstrating the same effectiveness of this protocol in comparison with modifi ed phenol-chloroform extraction protocol. This DNA extraction procedure constitutes an alternative and effi cient replacement for previous protocols for improving fi sh molecular studies.
J.A. Povh, N.M. Lopera-Barrero, R.P. Ribeiro, E. Lupchinski Jr., P.C. Gomes, and T.S. Lopes. 2008. Genetic monitoring of fi sh repopulation programs using molecular markers. Cien. Inv. Agr. 35(1):5-15. Native fi sh populations are decreasing due to several impact factors of the environment.In order to address this problem, several actions, for example stocking, have been accomplished. However, if these actions are undertaken without any scientifi c support, their effi ciency is low, and they can cause loss of genetic diversity in the populations, and irreversible impacts in the next generations. For this reason, the use of molecular markers to assess diversity through genetic monitoring, can contribute to the conservation of the icthyofauna.
Resumo -O objetivo deste trabalho foi avaliar a diversidade genética em quatro estoques de tambaquis (Colossoma macropomum) de diferentes regiões do Brasil, por meio de marcador RAPD. Foram utilizados 10 iniciadores para analisar 116 indivíduos, coletados de pisciculturas nos municípios de Urupá, RO, Teixeirópolis, RO, Neópolis, SE e Sorriso, MT. Foram encontradas diferenças nas frequências de 67 fragmentos, com um fragmento exclusivo em Sorriso e dois em Neópolis. Observaram-se altos valores de polimorfismo (72,92 a 83,33%), diversidade genética de Nei (0,27 a 0,30) e índice de Shannon (0,39 a 0,45). A análise da variância molecular, demonstrou que a maior parte da variação está dentro de cada estoque e não entre os estoques.A identidade e a distância genética entre os agrupamentos variou de 0,93 a 0,98 e 0,02 a 0,07, respectivamente, com menos distância entre os agrupamentos Urupá x Sorriso e entre Teixerópolis x Neópolis. A diferenciação genética variou de baixa a moderada (F st = 0,03 a 0,15) e o número de migrantes por geração foi alto (Nm = 5,96 a 24,3), entre os agrupamentos. Os estoques apresentam alta variabilidade e baixa diferenciação e distância genética entre si.Termos para indexação: Colossoma macropomum, diversidade genética, RAPD. Genetic variability of tambaqui (Teleostei: Characidae) from different regions of BrazilAbstract -The objective of this study was to evaluate the genetic diversity of four broodstocks of tambaqui (Colossoma macropomum) from different regions of Brazil using RAPD markers. Ten primers were used to analyze 116 individuals, collected from fish cultures of three municipalities in Brazil: Urupá, RO; Teixeirópolis, RO; Neópolis, SE; and Sorriso, MT. Differences in the frequencies of 67 fragments were found, with a unique fragment in Sorriso and two in Neópolis. High values of polymorphism (72.92 to 83.33%), Nei's genetic diversity (from 0.27 to 0.30) and Shannon index (from 0.39 to 0.45) were observed. Analysis of molecular variance showed that most of the variation is within each broodstock and not among them. The identity and genetic distance among the groups ranged from 0.93 to 0.98 and from 0.02 to 0.07, respectively, with less distance between clusters Urupá x Sorriso and Teixerópolis x Neópolis. Genetic differentiation ranged from low to moderate (F st = 0.03 to 0.15) and the number of migrants per generation was high (Nm = 5.96 to 24.3) among the groups. Stocks have high variability and low genetic differentiation and distance among them.
Resumo -O objetivo deste trabalho foi estudar a diversidade genética das amostras de pacu do médio Rio Paranapanema e do estoque de reprodutores utilizado no programa de repovoamento da Estação de Aqüicultura e Hidrologia da usina hidrelétrica Duke Energy, por meio do marcador RAPD. Genetic diversity of pacu from the Paranapanema River and from the broodstock of a stock enhancement programAbstract -The objective of this work was to study the genetic diversity of pacu samples from the middle Paranapanema River and from the broodstock used in the stock enhancement program of the Aquaculture and Hydrology Station at the Duke Energy Power Plant, by RAPD marker. Fourteen primers were used to analyze 30 individuals captured in the middle Paranapanema River and 29 individuals from the broodstock. The Shannon genetic diversity index and percentage of polymorphic fragments were higher in individuals captured in the Paranapanema River. Genetic similarity was larger in individuals of the broodstock. Results of analysis of molecular variance showed that the major part of the genetic variation is within the groups (84.2%) and not between them (15.8%). The identity and genetic distance among the groups were 0.9517 and 0.0549, respectively. Moderate genetic differentiation (F ST = 0.15) and high number of migrants per generation (Nm = 5.33) were observed between the two groups. The genetic diversity was lower in the broodstock than in fish captured in the middle Paranapanema River.
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