Isolamento e caracterização de locos de microssatélite para Neothraupis fasciata, (Emberizidae, Passeriformes, Aves),

AUTOR(ES)
DATA DE PUBLICAÇÃO

2009

RESUMO

Microsatellite molecular markers are small repeated sequences in tanden compound of 1 to 6 nucleotides. Widely used as genetic markers, the microsatellites have a particular attribute that is the high rates of mutation. These molecular markers or are widely used in conservation genetics, population genetics, forensic research, diagnosis of diseases, ecological studies and analysis of paternity. This range of applications are explained by the fact that microsatellites are co-dominant and multi-allelic, are highly reproducible, have high resolution and are based on polymerase chain reaction (PCR). Neothraupis fasciata is endemic of the Cerrado and occurs in areas with little human interference. These characteristics make this specie an ideal biological model to assess the possible effects on Cerrados fragmentation above the birds genetic diversity. Furthermore, this specie presents cooperative breeding system. The use of microsatellite molecular marker types will be important for kinship analysis to obtain new results on reproductive behavior of the species studiesand understand the populations genetic structure . On this study were developed and characterized microsatellites loci that were tested in eight otherbirds species. The microsatellites loci were developed from non-enriched genomic library. The library was constructed using the plasmid vector pCR4Blunt-TOPO using the TOP Shotgun Subcloning Kit. The purified inserts were sequenced using the T3 and T7 primers and the ET Terminator kit DYEnamic. The samples were analyzed in an automatic sequencer (MegaBACE 500). 928 clones were sequenced with a total of 278 inserts, of which 51 (18%) contained microsatellites loci, which are: 27 di, 17 tri, four tetra and three pentanucleotídeos. 17 primer pairs were designed and amplifications of sixteen these primer pairs had a clear interpretation in temperature ranging from 52 to 56 C and were characterized using 24 individuals of a population of N. fasciata. The microsatellite loci showed an average of 5.3 alleles per locus (ranging from 2 to 18), observed heterozygosity ranging from 0.083 to 1.0 (Ho = 0.394, mean for all loci) and expected heterozygosity ranging from 0.117 to 0.915 ( He = 0.540, mean for all loci). The loci had developed a high value of excluding paternity (0.999) and low probability of genetic identity (1.72x10-10) which makes it possible to use these loci on kinship analysis and species genetic structure . The transferability was tested in Sicalis flaveola, Ammodramus humeralis (Emberizidae), Thraupis sayaca, Gypsnagra hirundinacae, Paroaria capitata, Ramphocelus carbo (Thraupidae), Casiormis rufus (Tyrannidae) and Xiphorhynchus picus (Dendrocolaptidae).. Among the 16 synthesized primer pairs, 11 pairs amplified in S. flaveola, T. sayaca, G. hirundinaceae, P. capitata and R. carbo. The success of amplification may be useful on Passeriformes studies, especially for the family Emberizidae and Thraupidae species

ASSUNTO(S)

neothraupis fasciata emberizidae amplificação heteróloga microssatélites (genética); comportamento reprodutivo; cerrados thraupidae neothraupis fasciata microssatélite microsatellite cerrado outros cerrado emberizidae cross-species amplification thraupidae

ACESSO AO ARTIGO

Documentos Relacionados