Microsatellite markers obtained by 454-Roche pyrosequencing technology and its application for the study of Odontesthes bonariensis wild and hatchery populations.

VILLANOVA GABRIELA VANINA; VERA MANUEL; DIAZ JUAN; MARTINEZ PAULINO; CALCATERRA NORA BEATRIZ; ARRANZ SILVIA EDA

Villavicencio

Conferencia; IV CONFERENCIA LATINOAMERICANA SOBRE CULTIVO DE PECES NATIVOS; 2013

Universidad de los Llanos

Pejerrey (Odontesthes bonariensis) is one of the most popular inland fish in South America and has a long history of introductions in many countries. However, few genetics tools are available for it. Our aims were, to describe repetitive sequences of the pejerrey genome, identify and characterize microsatellites markers, and use them to compare genetic diversity of a wild and hatchery populations. Using 454-Roche pyrosequencing technology, a sample of pejerrey genome was obtained. In silico screening for repetitive sequences using available databases (CENSOR and RM database) was performed. The 25% of the screened sequences corresponded to DNA transposon (7.74%), LTR retrotransposons (3.93%), non-LTR retrotransposon (12.06%), unclassified (0.01%), small RNAs (0.48%), simple repeats (0.69%) and low complexity repeats (0.12%). Microsatellites we identified using Sciroko and MISA softwares. A sample of 13 polymorphic di-, tri-, tetra- and penta-nucleotides microsatellites was characterized. Wild and hatchery populations were analyzed (Table 1). Our initial analyses, although preliminary by the small size of the hatchery sample, suggested that genetic variability between these populations were significantly different. Monitoring hatchery strains and population genetics studies should be performed for an appropriate genetic resource management. Most of the microsatellites characterized in this work are a valuable tool for these purposes. Table 1. Genetic diversity of 13 polymorphic microsatellite loci from Odontesthes bonariensis in a wild and a hatchery samples. Wild population (42) Hatchery population (N= 12 ) Marker Na HO HE Ar F (Null) PHWE Fis Na Ho He Ar F(Null) PHWE Fis OdLAR8 4 0.310 0.305 2.455 -0.022 1 -0.016 2 0.250 0.228 1.998 -0.062 1 ¬0.100 OdLAR11 8 0.500 0.649 4.561 +0.122 0.054 0.231 4 0.500 0.707 3.833 +0.155 0.372 0.302 OdLAR12 2 0.024 0.024 1.238 -0.001 - - 1 0.000 0.000 1.000 - - - OdLAR16 21 0.524 0.920 11.591 +0.268 0.000** 0.433 4 0.583 0.572 3.831 -0.063 0.838 ¬0.199 OdLAR18 5 0.381 0.531 3.736 +0.141 0.061 0.285 2 0.636 0.524 2.000 -0.120 0.581 ¬0.228 OdLAR31 14 0.833 0.862 8.418 +0.012 0.169 0.034 6 0.583 0.822 2.000 +0.145 0.181 0.300 OdLAR38 24 0.881 0.953 13.382 +0.033 0.344 0.076 6 1.000 0.826 5.811 -0.120 0.939 ¬0.222 OdLAR40 5 0.143 0.400 3.299 +0.463 0.000** 0.645 2 0.083 0.083 5.935 -0.012 - - OdLAR43 3 0.071 0.070 1.660 -0.009 1.000 -0.016 2 0.083 0.083 1.833 -0.012 - - OdLAR44 3 0.167 0.378 2.834 +0.376 0.000** 0.562 3 0.000 0.507 1.833 +0.999 0.000** 1.000 OdLAR54 3 0.119 0.157 2.175 +0.126 0.025 0.246 2 0.167 0.159 1.978 -0.035 1.000 ¬0.047 OdLAR58 4 0.167 0.159 2.330 -0.034 1.000 -0.051 3 0.167 0.163 2.667 -0.034 1.000 ¬0.233 OdLAR59 3 0.048 0.047 1.476 -0.004 1.000 -0.006 2 0.083 0.083 1.833 -0.012 - - Na, number of alleles; HO observed and HE expected heterozygosity; Fnull Frequency of null alleles; PHWE, probability that the genotype population conformed to expectation probability to adjust Hardy Weinberg equilibrium are showed. (-) = Not Computed. (*)significant departure from HWE (P < 0.05), (**) significant after Bonferroni correction (p< 0.0025) ); Fis inbreeding coefficient estimated by Weir and Cockerham (1984); Ar; allelic richness.