Development Of Microsatellite Markers For Amur Sturgeon (Acipenser Schrenckii) And Its Application

Amur sturgeon (Acipenser schrenckii) is one of the28known fish species in theAcipenseriformes, which can be found in the Amur River basin in China and Russia. It is themost economic value in beluga sturgeon of rare fish with many advantages, such as, less disease,long life, grow faster. Chinese A. Schrenckii has become an important object and aquacultureindustry has occupies a certain position in Chinese fishery production. In this paper, the454pyrosequencing technique was used to sequence partial genome of A. schrenckii according to thestandard protocol. A total of127microsatellite primer pairs were designed and tested, and thesemarkers were used as follows: Screening the sex-related DNA markers through analyzing DNAdifferences between female and male; Analyzing the ability of these microsatellite primersamplifying in Acipenser baeri and Acipenser ruthenus; Analyzing DNA differences among ofAmur sturgeon (Acipenser schrenckii), kaluga (Huso dauricus) and their reciprocal hybrids. Themain results were summarized as follows.1. A total of127microsatellite primer pairs were designed and tested. Within the testedpopulation of30individuals,25markers produced unambiguous polymorphic products and35that successfully amplified were monomorphic. The remain67primer pairs resulted in poor orno amplification in A. schrenckii.2. There were9alleles derived from6SSR loci showed different distributions betweenfemale and male groups at a rate of>50%. Five alleles HLJSX194-239bp, HLJSX201-207bp,HLJSX209-210bp, HLJSX210178bp, and HLJSX215-221bp were found to be significantlyhigher frequency in the female samples than those in the male samples. However, there weresignificantly higher frequency in4alleles HLJSX194-184bp, HLJSX201-214bp,HLJSX210-233bp, and HLJSX226-194bp in the male samples than those in the female samples,indicating that these loci are linked with sex-determinate loci.3. We had analyzed the transferability of Amur sturgeon (Acipenser schrenckii) SSR primersin A. baeri and A. Ruthenus, the results showed that the efficient amplification ratio wererespectively68.75%and65.63%. The results of this research showed that the alleles andheterozygoties of A. Baeri is higher than that of A. Ruthenus, and we could use these new SSRmarkers analyzing genetic diversity of the sturgeon and add the number of A. baeri and A.Ruthenus by an economic method.4. Results showed that there were5primer pairs which could effectively identify the threepopulations. Among which, the specific loci from a part of A.schrenckii and H.dauricus could bedistinguished effectively in the amplified products of HLJSX226, HLJSX329, HLJSX332, HLJSX350, in addition, HLJSX350amplification products can be used to obviously distinguishthe characteristic loci of parents’population, and determine their reciprocal hybrids.