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Developing Microsatellite Multiplex PCR Panels for Topmouth Culter (Culter alburnus) and their Application in Parentage Assignment

Developing Microsatellite Multiplex PCR Panels for Topmouth Culter (Culter alburnus) and their Application in Parentage Assignment

Jianjun Fu1, Shili Liu2, 3, Wenbin Zhu1, Yongyi Jia2, 3, Lanmei Wang1, Yufen Yang4, Zhimin Gu2, 3*and Zaijie Dong1*

1Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture and Rural Affairs, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, Jiangsu, China 
2Key Laboratory of Healthy Freshwater Aquaculture, Ministry of Agriculture and Rural Affairs, Zhejiang Institute of Freshwater Fisheries, Huzhou 212001, Zhejiang, China 
3Key Laboratory of Freshwater Aquaculture Genetic and Breeding, Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou 212001, Zhejiang, China 
4Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, Jiangsu, China 
 
Jianjun Fuand Shili Liucontributed equally to this work.

*      Corresponding author: guzhimin2006@163.com; dongzaijie@ffrc.cn

ABSTRACT

The topmouth culter (Culter alburnus) is an economically important fish in China. However, natural stocks have been decreasing rapidly in recent years. Genetic improvement breeding programs of C. alburnus offer an essential strategy in developing a sustainable solution. In this study, we developed three multiplex PCR panels, including twelve microsatellite loci for C. alburnus, and applied parentage analyses using 36 candidate parents and 136 offspring from four independent artificial breeding pilots. Based on allele frequency analysis using 36 candidate parents, the number of alleles ranged from 7 to 22, observed heterozygosity ranged from 0.39 to 0.75, and expected heterozygosity ranged from 0.57 to 0.91. All loci were highly informative (polymorphic information content, PIC >0.5). We detected a significant correlation between exclusion probabilities for parent pairs (E-PP) and PIC (P<0.01). Simulation analysis revealed that a high assignment rate (>95%) was achieved when the number of candidate parents was less than 200. Meanwhile, real parentage analysis revealed that almost each offspring (135/136, 99.26%) was unambiguously assigned to a parent pair with high accuracy (100%), respectively. In addition, we detected significantly unequal progeny contributions of parents and parent pairs in total or in each artificial breeding pilots (P<0.01). In brief, we developed microsatellite multiplex PCR panels for C. alburnus with high capability of monitoring pedigree information, which can be used as a basic tool for family selection breeding of this species. 
 

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Pakistan Journal of Zoology

February

Vol. 53, Iss. 1, Pages 1-400

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