Genetic Diversity Analysis Of Rough Skin Sculpin (Trachidermus Fasciatus) In Native Populations Of China

In this paper, we compared and analyzed the DNA molecular marker technologiesin the research of genetic diversity of aquatic species, including RFLP, AFLP, RAPD,SSR, and ISSR. We discussed the advantage and disadvantage of various DNAmolecular marker technologies in the application and aimed to find out the mostappropriate method to research the genetic diversity of aquatic organisms.The research object of this paper is Trachidermus fasciatus, Trachidermusfasciatus as a national level2to protect the wild animals, there has the importantposition and value in the historical culture and genetics in all. We have done a lot ofwork, finally, we found four Trachidermus fasciatus wild natural group in China, andhas its own hatchery exclusive laying eggs, the groups are respectively zhejiangfuchunjiang group, the mouth of the Yellow River in shandong, hebei qinhuangdaoluanhe river community groups and liaoning the yalu river group. Then arrived to eachdistrict living sampling respectively, and get the right to Trachidermus fasciatus fins onalcohol article dehydration save, extraction method-genomic DNA by Method ofPhenol-Chloroform.And then use a single impact factor of the screening method established the fourTrachidermus fasciatus group ISSR-PCR program, and through the optimized selectionfor: reaction system20μL: including DNA template1μL(about30ng), ISSR primer1μL (0.4μ mol/L),10by PCR Buffer2μL (Mg22.5mmol/L), dNTP1μL(250μmol/L), Taq enzyme0.2μL (5U/μ L), ddH2O14.8μ L. Polymerase chain reaction(PCR) program:95℃the degeneration3minutes, then94℃30seconds, annealingtemperature45seconds (table2-1),72℃extensions60seconds,40cycles,72℃extension5minutes,4℃preservation. Use this reaction system for100of a completeset of primers ISSR annealing temperature on the exploration, and eventually selected15best polymorphism primers, Trachidermus fasciatus for the ISSR analysis lay thefoundation.The inter-simple sequence repeat(ISSR) was conducted to analyze the genetic variation of four native populations of Trachidermus fasciatus in China Zhejiangpopulation(Z), Shangdong population(S), Hebei population(H) and Yalvjiangpopulation(Y), with15polymorphic primers selected from100ISSR primers. For eachpopulation,30samples were used in the study. Six to seventeen bands were amplifiedfor each primer. Altogether,181useful loci were detected and out of them,136oneswere polymorphic, accounting for75.14%. Among the four populations, H populationhad the lowest effective allele number (1.2585), Nei’s gene diversity(0.1530),Shannon’s information index(0.2322), and polymorphic loci percentage(49.17%), whileY population’s indexes (1.4794,0.2777,0.4112, and74.03%, respectively) were thehighest. The corresponding values of Z and S populations were higher than these of Hpopulation and lower than these of Y population. If took the four populations as awhole, the total Nei’s gene diversity and Shannon’s information index were0.2691and0.4009, respectively, showing a relatively high diversity of T. fasciatus. The values ofGSTand Nm were0.1313and3.3094, indicating middle differentiation and some degreeof gene exchange existed among the populations. As results of AMOVA indicated, therewere85.54%variation existed in the individuals and14.46%variation amongpopulations. In the same time, there was significantly differentiation among the fourpopulations (P<0.05or P <0.01),too. In the UPGMA tree conducted on the geneticdistance, Z and S populations clustered firstly, then with Y population, and finallyjoined to H population. Obviously, the Z, S, and Y populations had a relative closerelationship according to their geographic distance, whereas H population showed cleardivergence to the other three populations.The object of study is currently in China found four Trachidermus fasciatus wildgroup, the total of120sample, the sample size is big enough, and geographical positiondistribution from the south to the north is quite comprehensive, for China’s survival andTrachidermus fasciatus genetic diversity status and to provide detailed and accurate enoughbased data. In addition also established the Trachidermus fasciatus ISSR-polymerase chainreaction (PCR) system and the future of Trachidermus fasciatus group molecular geneticsresearch is more ISSR pertinence and reference value. This paper analyzed the results showthat the Trachidermus fasciatus four wild groups the genetic structure of the geneticdiversity and current situation, this also is Trachidermus fasciatus germplasm resourcesprotection, development and utilization provide basic data for the hatch and rearingTrachidermus fasciatus and lays a foundation, at the same time as the national secondary Trachidermus fasciatus protect animal research accumulated theory basis.