59º Congresso Brasileiro de Genética
Resumos do 59o Congresso Brasileiro de Genética • 16 a 19 de setembro de 2013
Hotel Monte Real Resort • Águas de Lindóia • SP • Brasil
www.sbg.org.br - ISBN 978-85-89109-06-2
5
MOLECULAR TECHNIQUES APPLIED
IN THE STUDY OF KINSHIP ANALYSIS
OF PODOCNEMIS SEXTUBERCULATA
(TESTUDINES, PODOCNEMIDIDAE)
Miguel, GSS1,2; Farias, IP³ ; Fantin, C1,4
Escola Superior de Ciências da Saúde, UEA, Manaus, AM; 2Bolsista CNPq - Programa Institucional de Bolsas de Iniciação
em Desenvolvimento Tecnológico e Inovação (PIBITI); 3Universidade Federal do Amazonas – UFAM; 4Mestrado em
Biotecnologia e Recursos Naturais, UEA, Manaus, AM
1
[email protected]
Keywords: Podocnemis; paternity; microsatellite, polyandry, genotyping.
Turtles appeared in Mesozoic Era, over 200 million years ago and are considered the oldest existing reptiles. They are
scattered among various places such rivers, lakes and swamps. The order Testudines (turtles and tortoises) is divided
into 13 families and the present study focus on the specie Podocnemis sextuberculata, Podocnemididae. This turtle
inhabits clear water rivers, is widely consumed in Amazon region and its clandestine trade is a predation factor. This
study aimed to understand the reproductive behavior (by determining the paternity type) of this specie in order to
contribute for the elaboration of further preservation studies. Study site was at Trombetas (Trumpets) River - PA,
where during spawning time six nests of P. sextuberculata were identified and marked. After eggs hatching, 100 mL
of blood from each offspring was collected, totalizing sixty samples. DNA extraction followed the CTAB method.
Extracted DNA was subjected to polymerase chain reaction (PCR) following the economic protocol described by
Schuelke, using three microsatellite loci (Puni_1D12, Puni_1H9 and Puni_2A9) developed for Podocnemis unifilis.
Amplification of microsatellite region was checked by 1% agarosis gel electrophoresis. Genotyping of amplified
microsatellite loci was performed by an automatic sequencer (ABI 3130 xl) which provided data for determining
the genotype of each offspring. Kinship level between individuals from each nest was determined by a simple alleles
counting. As result, we obtained the inference of multiple paternity in the six nests studied. Alleles number ranged
between 4 to 14, where locus 1H9 was the least polymorphic (in all studied nests) whereas locus 1D12 was the
most polymorphic (nest 1). Alleles counting of offsprings shows that nest 1 was the one with major contribution
of males (6 individuals). It is concluded, therefore, that P. sextuberculata shows polyandry as main characteristic
of mating behavior, where one female has intercourse with several other males, which is supported by our results.
Financial Support: CNPq and the Fundação de Amparo à Pesquisa do Estado do Amazonas (FAPEAM)