Analysis Of Genetic Diversity And Whole-Genome Sequence In Ussurian Wide Pig(Sus Scrofa Ussuricus)

With the rapid development of molecular biology technology,it is possible to analyze the genetic diversity and genome sequence of wild animals.There are 5 genera.8 species and 23 subspecies of wild boar in the world.In China,there are 1 specie and 7 subspecies of wild boar resources.According to the geographical location,wild boar in Heilongjiang Province should belong to the Northeast subspecies of wild boar in China(Sus scrofa ussuricus).Wild boar is an important wild resource.The research on the genetic diversity and adaptive evolution of the ussurian wide pig can not only obtain the basic heritage data information of the ussurian wide pig,enrich the research on the genetic diversity of the ussurian wide pig,provide the basis for other studies in the follow-up group,but also greatly enriches the database of genetic variation sites of wild boar genome,and provides important basic scientific data for the study of genetic mechanism of germplasm characteristics and the mining of excellent gene resources of ussurian wide pig.In this project,the genetic diversity and genome sequence of ussurian wide pig were researched.Through the molecular biological identification of ussurian wide pig,genetic diversity analysis based on mitochondrial gene and whole genome re sequencing,and analysis of selected genes,the conclusions are obtained:1.Identification of ussurian wide pig and domestic pig by PCR-RFLP.The polymorphism.of MC1R gene was detected by Acc? and BspHI.The result showed that BB/AA was a unique genotype combination of ussurian wide pig2.The 1 137bp coding region of cytochrome b gene of ussurian wide pig was obtained.The phylogenetic tree of wild boar in different regions of Europe.Asia,Africa and China was constructed.The results showed that the genetic relationship was basically consistent with the geographical distribution characteristics.The mtDNA D-loop(1 50bp-591bp)of ussurian wide pig was sequenced.The molecular evolutionary tree based on D-loop region was constructed,including wild boar in Heilongjiang.Jilin,Liaoning,Inner Mongolia and South Korea.The results showed that the genetic relationship was basically consistent with the geographical distribution characteristics.21 haplotypes were obtained from mtDNA D-loop in Eurasian wild boar,among which 8 haplotypes were found in ussurian wide pig.The results of constructing haplotype composition ratio and cluster diagram support the dualism view of the origin of two female lines in Europe and Asia.3.A total of 36.28GB data were obtained from the whole genome resequences in ussurian wide pig.10 275 059 SNPs and 1 071 744 indels were found,including 35 946 missense mutations,involving 9 739 functional genes and 7 739 173 SNPs.The complete genome sequences of 80 individuals including Asian wild boar,Asian domestic pig,European wild boar,European domestic pig and Warthog were downloaded from NCBI database,and the molecular evolutionary tree based on SNPs information of these 80 individuals was constructed.It was found that European pig and Asian pig were obviously clustered on two different branches,while ussurian wide pig was clustered on the branch of Asian pig,and compared with Chinese local pig and Japanese wild pig,the results of principal component analysis were consistent with the results of principal component analysis.4.The results showed that 29.81Mb of the ussurian wide pig genome was selected from the 4 resequenced data,involving 411 genes,among which 279 genes with functional annotation were involved.Go analysis was used to obtain the enrichment results of cell components,molecular functions and signal pathways.The protein interaction network was constructed by 279 genes with functional annotation,and 147 genes formed interaction relationship.16 genes related to nervous system development were selected and identified by qPCR.The results showed that the expression of these genes was significantly different between the ussurian wide pig and Min pig.5 genes related to immunity,reproduction and nervous system were sequenced,and a lot of genetic diversity data were obtained to support the next research.