Isolation Of Microsatellite Makers Of Dabry’s Sturgeon And Application In Its Parentage Identification

Dabry’s sturgeon, Acipenser dabryanus, is a resident freshwater fish of Chinese endemic species. Due to the effects of human activities, the population size of Dabry’s sturgeon has been decreased sharply. The species was listed as the First Class Protected Animal by the Chinese Government in1988and also characterized as a Critically Endangered species in the International Union for Conservation of Nature and Natural Resources (International Union for Conservation of Nature and Natural Resources, IUCN). Therefore, it is necessary to take some effective measures to protect Dabry’s sturgeon, such as artificial domestication, propagation, enhancement and so on. In this study, microsatellite primers were developed in Dabry’s sturgeon, which is helpful to genetic testing, Genetic Linkage Map construction and assessment of enhancement effect. Subsequently, paternity testing of Dabry’s sturgeon was performed by microsatellite makers obtained to optimizate artificial breeding technology. This research will design the population genetic management strategy for avoiding inbreeding and offer a reference for other sturgeon paternity test.1.(CA)15and (CT)15enriched microsatellite libraries in A. dabryanus were constructed by using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol. Sixty-seven microsatellite primers were designed for PCR amplification of twenty-four individual. As a result, twenty-six polymorphic microsatellite markers were developed. The polymorphism analysis showed that the number of alleles of microsatellite loci ranged from3-13, with an average of5.2. Mean expected heterozygosities (HE) and Shannon-Wiener Diversity Indices (H’) per locus ranged from0.303to0.806, and from0.480to1.893, respectively.2. Because of the generality of the closely related species microsatellite markers, we found thirteen common microsatellite primers in the microsatellite library of Lake sturgeon and shovel sturgeon; meanwhile, forty primers were designed based on the published Chinese the sturgeon EST sequences. A total of fifty-three microsatellite markers were used to the PCR amplification of Dabry’s sturgeon genomic DNA. Finally, fifteen high polymorphism primers were got28%utilization ratio.3. Among41polymorphic primers in Dabry’s sturgeon microsatellite library and the related species, twelve primers were choose to synthesis labeled with fluoresce. STR genotyping was used to construct the parent database after the PCR amplification with two pairs of parents and offspring DNA as templates. Paternity testing technology was proved successfully on Dabry’s sturgeon research. Find the offspring corresponding parents by exclusive method on the comparison analysis for every microsatellite loci alleles of the individual offspring. The study identified successfully the parents of six offspring.