Nuclear DNA markers for identification of Beluga and Sterlet sturgeons and their interspecific Bester hybrid

Abstract

Sturgeons (Acipenseriformes) are among the most endangered species in the world due to fragmentation and destruction of their natural habitats and to overexploitation, mainly for highly priced caviar. This has led to the development of sturgeon culture, originally for reintroduction, but more recently for caviar production. In both cases, accurate species identification is essential. We report a new tool for accurate identification of Huso huso and Acipenser ruthenus based on nuclear DNA markers. We employed ddRAD sequencing to identify species-specific nucleotide variants, which served as specific binding sites for diagnostic primers. The primers allowed identification of Huso huso and Acipenser ruthenus as well as their discrimination from A. baerii, A. schrenckii, A. gueldenstaedtii, A. stellatus, A. persicus, A. mikadoi, A. transmontanus, and H. dauricus and identification of A. ruthenus and H. huso hybrids with these species, except hybrid between A. ruthenus and A. stellatus. The species-specific primers also allowed identification of bester (H. huso × A. ruthenus), the most commercially exploited sturgeon hybrid. The tool, based on simple PCR and gel electrophoresis, is rapid, inexpensive, and reproducible. It will contribute to conservation of remaining wild populations of A. ruthenus and H. huso, as well as to traceability of their products.

A new marker, isolated by ddRAD sequencing, detects Siberian and Russian sturgeon in hybrids

“…The positive result for a single Amur sturgeon and, conversely, lack of amplification in three Siberian and eight Russian sturgeons is probably due to genetic admixture, as also observed by Boscari et al . The marker, combined with others already available (Table ), will contribute to enforcing controls against fraud and illegal trade of sturgeon products. Moreover, the ability to detect hybridization with Russian or Siberian sturgeon is relevant for ex situ conservation programs given that hybridization is considered an immediate genetic threat to endangered populations …”

“…Sixteen samples each of sterlet, Siberian sturgeon and beluga were analyzed by ddRAD sequencing described previously (Appendix ). At positions 33 and 34 of a 110‐bp ddRAD fragment, an AG variant was observed in all 655 reads of the Siberian sturgeon, whereas a CA variant was observed in all 410 reads of the sterlet and in all 505 reads of the beluga (Fig.…”

Application of interspecific Somatic Cell Nuclear Transfer (iSCNT) in sturgeons and an unexpectedly produced gynogenetic sterlet with homozygous quadruple haploid

“…The larvae of most sturgeon species can be unambiguously identified only by appropriate molecular markers 45 . To uncover, whether NT-fish contains genome of Russian sturgeon donor, we accommodated recently developed nuclear DNA markers 46 . The results of molecular genotyping clearly showed that NT-fish contains only sterlet genome.…”

“…Samples were tested by sterlet positive primer pair 247_AR + 247_uni and subsequently by sterlet negative primer pair 247_ARn + 247_uni as described in Havelka, et al . 46 . If a sample contains only sterlet DNA, the 247 bp fragment is amplified only by sterlet positive primer pair.…”

Using machine learning algorithms to analyse the scute structure and sex identification of sterlet Acipenser ruthenus (Acipenseridae)

“…Above methods have difficulties with mass application in aquaculture and can have a high error (the first two methods) or can traumatize fish (the last four methods). Furthermore, a sturgeon sex gene has not been found, unlike most fish species (Fopp‐Bayat, ; Keyvanshokooh & Gharaei, ; Khodaparast, Keyvanshokooh, Pourkazemi, Hosseini, Keyvanshokooh, Pourkazemi, Hosseini, & Zolgharnein, ; Vizziano‐Cantonnet, Di Landro, & Lasalle, ); however, a study in this area are still under way (Burcea et al, ; Havelka, Fujimoto, Hagihara, Adachi, Fujimoto, Hagihara, Adachi, & Arai, ; Vizziano‐Cantonnet et al, ; Vizziano‐Cantonnet, Lasalle, DiLandro, Klopp, & Genthon, ). Currently, the ultrasound diagnostics is the most successful method of early sturgeon sex determination (Chebanov & Galich, ; Memiş et al, ; Webb et al, ) and can determine the sex starting from 2 years for Sterlet sturgeon ( Acipenser ruthenus ), from 2–3 years for Siberian ( A. baerii ) and Russian ( A. gueldenstaedtii ) sturgeons, from 3–4 years for White sturgeon ( A. transmontanus ) and from 4–5 years in Beluga ( Huso huso ) (Chebanov & Galich, ).…”