Analysis Of Genetic Structure Of Sable (Martes Zibellina L.1758) Based On Microsatellite

The sable(Martes zibellina),belonging to the Carnivorous Mustelidae family,is widely distributed in coniferous forests or coniferous and broad-leaved mixed forests in Eurasia and surrounding islands.Due to the wide distribution of sable,the large variation in morphological characteristics and the reintroduction phenomena,the division of sable subspecies becomes a problem that has plagued relevant scholars.Russia,China and Japan are the main habitats of sable.The number of individuals in sable is relatively high.The genetic diversity,genetic differentiation and genetic structure of different geographic populations in three countries have always been the research hotspots of relevant scholars.However,there are many problems in the related studies,such as fewer geographical populations or smaller population distribution areas,fewer sample sizes in different populations,lack of multi-samples,and large-scale geographical multi-group research results.This study was based on simplified restriction-site-associated DNA(RAD)sequencing of genomic restriction sites,and the species-specific microsatellite locus was obtained by primer selection.These molecular markers were used to study genetic structure of eight subspecies(M.z.obscura,M.z.sajanensis,M.z.linkouensis,M.z.zibellina,M.z.arsehjevi,M.z.yenisee nsis,M.z.yeniseensis,M.z.hamgyenensis,M.z.princeps)from northeastern China,Mongolia,and Russia.In the preselected 101 pairs of microsatellite primers,15 effective micro satellite loci were finally obtained,5 of which were highly polymorphic,and 2 loci belonging to low polymorphism.The population genetic diversity of 139 samples in 8 subspecies of sable was studied by screening microsatellite loci.The average number of alleles in the total population was 4.867 and the average number of effective alleles was 1.928.Allele distribution was uneven.The average observed heterozygosity of the eight subspecies populations at the 15 sites was 0.365,and the average expected heterozygosity was 0.426.The overall genetic diversity of the population was not high.Among the eight sub-populations,M.z.princeps subspecies had the highest genetic diversity,with the highest number of alleles being 3.867;followed by the M.z.zibellina subspecies,the number of alleles was 3.733;the number of alleles observed in the M.z.arsehjevi species was the lowest(1.800).The observational heterozygosities of M.z.princeps subspecies and M.z.obscura subspecies were high in eight subspecies,0.400 and 0.437 respectively.The F-Statistical of sable population was tested.In the general population,the mean Fis value was 0.029 and the mean Fit value was 0.151.The coefficients of genetic differentiation among the subspecies were within the range of 0.005 to 0.157,that is,there were different degrees of genetic differentiation among different subspecies,among which the genetic distance between M.z.arsehjevi subspecies and the other 7 subspecies was farthest.The gene flow between subspecies varied in size,with a maximum of 48.769 between theM.z.zibellina subspecies and M.z.princeps subspecies.The gene flow between the M.z.zibellina subspecies and M.z.ajanensis subspecies was 17.736,and the gene flow between other subspecies ranged from 1.343 to 16.757.Phylogenetic tree of 8 subspecies was constructed.Eight subspecies were divided into four clades:M.z.zibellina subspecies,M.z.princeps subspecies,M.z.sajanensis subspecies and M.z.yeniseensis subspecies were clustered into the first clade;M.z.linkouensis subspecies and M.z.hamgyenensis subspecies formed second clade;the third clade was M.z.obscura subspecies;the fourth clade was M.z.arsenjevi subspecies.Bayesian clustering analysis revealed that sable population was divided into two major taxa.For the analysis of 16 geographic populations,the group was divided into 6 major clades.The Arhara Rive group was listed as an independent clade.The WeiHe and ShangZhi groups were clustered together.The three groups were closest to their common ancestor.The LuoBei group was a separate clade.The WangQing,YiChun,and MoHe groups were clustered together.The Chikoi River group was an independent clade.The Tuqiang group and the Russian population were clustered together.