Development and characterization of a duplex PCR assay in Chinese sturgeon (Acipenser sinensis) for genetic analysis

Abstract

chinese sturgeon (Acipenser sinensis) has been listed as a critically endangered species on the iUcn Red List and is an endemic fish of China. Five sets of duplex polymerase chain reactions (PCR) assays were developed with 10 tetranucleotide microsatellites for Chinese sturgeon. The size of CS57, ZHX43, ZHX69, AS105, ZHX51, AS074, ZHX2, AS078, AS026 and AS073 products in 184 Chinese sturgeon individuals ranged from 257-305, 191-241, 251-285, 172-244, 236-260, 169-209, 194-234, 92-176, 165-257 and 120-164, respectively. The observed allele number of the 10 microsatellites ranged from 7 to 16, and the total number of alleles was 106. The number of alleles per individual in CS57, ZHX43, AS105, AS074, AS078 and AS026 was 1-4. The number of alleles per individual in ZHX69, ZHX51, ZHX2 and AS073 was 2-4. The mean number of alleles per locus per individual ranged from 2.01-3.76. The expected heterozygosity (H E), observed heterozygosity (H O), polymorphic information content (PIC) and Shannon-Weiner diversity index (H′) ranged from 0.582 to 0.899, from 0.676 to 1, from 0.518 to 0.886 and from 1.034 to 2.34, respectively. Despite many advantages, the use of microsatellites as genetic analysis tools can be limited by the cost of the associated experiment. To solve this problem, this set of five duplex PCRs will provide tools that are more helpful, less expensive and less time consuming than others used for genetic analyses in Chinese sturgeon. Chinese sturgeon (Acipenser sinensis) is an endemic and critically endangered species in China. Chinese sturgeon was once widely distributed in the Yangtze River, Pearl River and Chinese seas 1. The wild population of Chinese sturgeon has fallen drastically in the past decades due to human activities, such as the use of pesticides, industrial overfishing and environmental pollution 2-4. Currently, the wild population of Chinese sturgeon in the Yangtze River is very small. The need for species rehabilitation is known, and efforts have been made to protect it. To prevent this species from becoming extinct, artificial propagation and tagged ranching have been performed every year since 1984 5,6. A national reserve was established, with the purpose of breeding and restocking Chinese sturgeon. The first artificial propagation of cultured Chinese sturgeon was successfully performed in 2009 by the Chinese Sturgeon Research Institute, China Three Gorges Corporation (Yichang, China). The success of artificial propagation in Chinese sturgeon is of significance for the protection of this species. Although, many studies have been performed to support the recovery of this species, the number of Chinese sturgeons is becoming increasingly small. The Chinese sturgeon is listed as a Critically Endangered species by the International Union for Conservation of Nature and Natural Resources Red List (IUCN Red List) 7 and is a first class protected animal in China 8. Genetic investigations play an important role in protecting Chinese sturgeon and can prevent inbreeding during the culture of this species....

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