Cross-amplification of heterologous microsatellite markers in Piracanjuba

Abstract

Brycon orbignyanus, popularly known in Brazil as piracanjuba, is a fish with great economic value but whose natural population drastically decreased in number during the last years. In this context, genetic variability studies of natural stocks and in restocking programs are fundamental for the adoption of conservation measures. Current analysis verifies the cross-amplification of heterologous primers in B. orbignyanus. Fifty-two primers of the species Brycon opalinus, Brycon hilarii, Brycon insignis, Prochilodus sp., Piaractus mesopotamicus, Colossoma macropomum and Oreochromis niloticus were tested. Primers with the best reproducibility were applied to a sample of 20 individuals and the genetic parameters were calculated. Nine primers provided good results for cross-amplification with B. orbignyanus, involving (BoM5 and BoM13) of Brycon opalinus, (Bh5, Bh6, Bh8, Bh13 and Bh16) of Brycon hilarii, (Bc48-10) of Brycon insignis and (Par80) of Prochilodus argenteus. Primers of Piaractus mesopotamicus, Colossoma macropomum and Oreochromis niloticus failed to provide amplification or provided non-specificity. Results demonstrated the possibility of using primers of different species and genera of B. orbignyanus, facilitating genetic studies on the species.

Transferability of heterologous microsatellite primers in Brycon gouldingi

“…According to oLiVeirA et al (2006), the transferability between related species depends on the degree of conservation of the primer binding regions in the DnA, with evolutionarily closer species being more successful in cross-amplification. recent studies have demonstrated this by evaluating the crossamplification of several species of the neotropical fish B. orbignyanus (cArmo et al, 2015;cAsTro et al, 2017), obtaining greater amplification predominantly among species of the same genus, which is consistent with the results of this study.…”

“…The use of microsatellites requires specific primers for each species, which are currently not available for B. gouldingi. however, several studies have shown that pairs of primers designed for one species can be used for other species of the same genus (oLiVeirA et al, 2006;LoPerA-bArrero et al, 2016a;cAsTro et al, 2017), which are commonly known as heterologous primers. Therefore, studies aimed to characterize these primers required for genetic studies of this species.…”

Genetic diversity of migratory fish populations of the Rio Grande Reservoir (São Paulo, Brazil)

“…Similarly, the Ho values of the P. mesopotamicus population was in accordance with that found in Pantanal populations of the same species (CALCAGNOTTO;DESALLE, 2009). Finally, the Ho of B. orbignyanus was also similar to that found by other studies for natural continuation stocks (ASHIKAGA et al, 2015) or captive stocks (CASTRO et al, 2017). However, B. orbignyanus requires special attention since it is highly sensitive to environmental changes, which makes it endangered in certain regions (ASHIKAGA et al, 2015).…”

“…It is important to emphasize that, for this species, heterologous primers of related species were used. These primers were previously tested and produced good amplification and reproducibility standards, agreeing with earlier research (CASTRO et al, 2017).…”

“…For P. lineatus DNA samples, PCR was performed according to the methodology described by Souza et al (2018), using eight microsatellite loci: Par12, Par14, Par21, Par43, Par80 (BARBOSA et al, 2006(BARBOSA et al, , 2008, Pl01, Pli30, and Pli60 (YAZBECK; KALAPOTHAKIS, 2007). For B. orbignyanus, PCR was performed according to Castro et al (2017), using eight microsatellite loci: BoM5, BoM13 (BARROSO et al, 2003), Bh5, Bh6, Bh8, Bh13, Bh16 (SANCHES; , and Par80. Finally, the amplification conditions for P. mesopotamicus was according to the methodology described by Ribeiro et al (2015), using seven loci: Pme2, Pme4, Pme5, Pme14, Pme20, Pme28, and Pme32 (CALCAGNOTTO et al, 2001).…”