Development Of Microsatellite DNA Markers And Population Genetics For Sand Lance (Ammodytes Personatus) And Roughskin Sculpin (Trachidermus Fasciatus)

MicrosatelliteDNA markers for Sand lance (Ammodytes personatus) and roughskinsculpin (Trachidermus fasciatus) were isolated developed by enrichmentlibraries-colony hybridization and PCR screening. Population structure and geneticdiversity were using these microsatelliteDNA markers in the two species.1. Isolation and characterization of microsatellite markers for two marine fishes(1) Isolation and characterization of microsatellite markers for Sand lance (A.personatus)Microsatellite enrichment library of Sand lance (A. personatus) was constructed,which was digested by HaeⅢ. The primers flanking31microsatellites isolated fromthe genomic library enriched for (AC)nand (AG)nwere designed in the Sand lance (A.personatus). Eleven primer pairs provided clear and polymorphic amplificationproducts. Based on characterization with36Sand lance individuals, the number ofalleles ranged from8to29. The values of HOand HEvaried from0.441to0.886and0.818to0.965, respectively. These markers are therefore potentially useful for studiesof the population genetics of the species.(2) Isolation and characterization of microsatellite markers for roughskin sculpin (T.fasciatus)Different from Sand lance (A. personatus), PCR screening after genomic libraryenriched constructed were used to develop14novel polymorphic and informativemicrosatellite markers for the roughskin sculpin (T. fasciatus). We characterized theseloci by genotyping29individuals in Wendeng population. A total of113alleles werefound. The number of alleles ranged from2to13across the14microsatellite loci. The values of HOand HEvaried from0.682to1.000and0.853to0.953, respectively.2. Molecular phylogeography and Genetic diversity for sand lance (A. personatus)The Japanese sand lance A. personatus is a common and commercially importantspecies in northwestern Pacific. Sand lance, A. personatus, usually constitutes a highproportion of the fish biomass in the regions where they occur. Microsatellite DNAmarkers were used to estimate the population structure and demographic history ofSand lance. Two high distinct lineages were detected in318individuals from16populations across the range of the species based on sequence variations ofmicrosatellite. The majority of individuals of the Dalian(Dl)、 Qingdao(Qd)、Fukuoka(Fuku)、Kagawa (Kag)、Ise Bay (Ise)、Hyogo (Hy)、Kashima(Kas)、Otsuko(Ot)、Sendai Bay Small (Ss)、Fukaura Kuka populations of sand lance were assignedto an inferred cluster, and above the93%of the individuals of the fourpopulations(Mutsu Bay (Mu)、Sendai Bay Large (Sl)、Hachinohe (Ha)、Ishikari Bay(Ish)、Rebun Island (Re)、Cape Soya (Ca) of sand lance were assigned to the other.Pleistocene coastal glaciations might have divergent in the Sea of Japan and Pacificcostal waters of Japanese Island, during the low sea level. Two major ecologicalgroups of sand lance may be accomplished since then. The high genetic diversity wasfound for sand lance based on microsatellite DNA markers. The large effectivepopulation size coupled with the high mutation rate may be responsible for the highgenetic diversity.3. Bottleneck effect and population estimate for roughskin sculpin (T. fasciatus)Roughskin sculpin (T. fasciatus) is a Catadromous species. The distribution area ofthe species in China has been reduced greatly and almost disappears completely alongthe sea areas in the east and the south caused by environmental pollution andover-catching. It is urgent for scientific management for sustainable exploitation. Toexamine population genetic diversity and structure of T. fasciatus, samples from fivedifferent locations of China and Ariake population in Japan were analyzed by usingmicrosatellite technology. The Polymorphism Information Content (PIC) were high(PIC=0.683-0.928). The expected heterozygosities ranged from0.763(C79-E) to0.970(E95-E). The observed heterozygosities varied from (HO)0.333(C44-E) to 1.000(C46-E、C44-QTJ). An effective population size reduction (bottle effect) isdetected in Te and Qt by Mode shift test. The effective population size of Wd and Tepopulation was below4000, and smaller than other populations.High significant genetic differentiations were found among6populations byscanning9microsatellite loci variations. The pairwise FSTvalues ranged from0.0131to0.0467. The majority of individuals (>95%) of the Ariake populations of roughskinsculpin were assigned to an inferred cluster, and above the93%of the individuals ofthe four populations of roughskin sculpin were assigned to the other. Current and itslife cycle might restrict the gene flows between populations.