Abstract Abstract Mandarin fish (Siniperca chuatsi) is one of the most important cultured freshwater fish in China. In order to carry out pedigree analysis of selective mandarin fish, compare identification effects of double digest restriction-site associated DNA sequencing (ddRAD-seq) and microsatellites, and establish paternity identification in mandarin fish, 30 pairs of microsatellite primers were selected from GenBank, and 11 of them with good specificity and polymorphism were screened by polyacrylamide gel electrophoresis. Then, these 11 primers were used to amplify 4 full-sib families whose pedigree information was known. Capillary electrophoresis was used to carry out microsatellite genotyping. The result of cluster analysis showed these four full-sib families were divided into four groups using the 11 pairs of microsatellite primers. All 3 283 SNP markers were obtained by ddRAD-seq, and they were used to construct a clustering tree, including 155 samples from the same family and 12 samples from other families. Finally, the 12 samples were effectively distinguished. The above 11 pairs of microsatellite primers were used to detect the 12 samples from other families, eight samples from the same family and their parents, and clustering tree was constructed. The result showed that the 8 samples from the same family and their parents were clustered together, while the 12 samples were clustered in the other cluster. These results illustrated that these 11 pairs of microsatellite primers had good family identification ability. Then, parentage determination of 126 offsprings, which were bred by 7 male and 13 female breeding parents, were carried out by these microsatellite loci. Simulation analysis showed that the non-exclusion probability of each locus ranged from 0.276 to 0.895, and the highest calculate exclusion probability was 99.999%. Among these 126 offsprings, 105 offsprings had been successfully identified for their pedigree relationship by parentage analysis and the success rate was 83.3%. These results illustrated that these 11 pairs of microsatellite primers had high calculate exclusion probability and good identification effect in pedigree identification. The paternity identification method established in this study will be helpful for breeding of mandarin fish.
ZENG Qiang-Kai,XUN Cheng-Fei,DONG Jun-Jian, et al. Establishment and Utilization of Paternity Identification in Mandarin Fish (Siniperca chuatsi) Using Microsatellites[J]. , 2017, 25(6): 976-984.
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