Population Genetic Structure Of Sika Deers (Cervus Nippon) And Application Of Molecular Markers In Identification For Imported And Exported Animal Products In Customs

This thesis includes four chapters:1. The progress of the study on evolution and conservation genetics of Cervidae was reviewed.2. Three hundred and fifty-one base pairs (bp) of the mitochrondrial control region from 39 sika deers (Cervus nippori) collected from Jilin (Northeast China population), Sichuan (Sichuan population), Anhui and Jiangxi (South China population) provinces were sequenced. Twenty-three variable sites were determined and five haplotypes were identified. The analysis of molecular variance (AMOVA) showed a significant genetic structure for sika deers in China Mainland (Ost=0.68, Fst=0.64, P<0.001). Nested clade analysis (NCA) generated almost identical results, supporting to treat these populations as different management units. No shared haplotype was found among China Mainland, Northern Japan, and southern Japan populations, and 25 fixed diagnostic sites differences separate the three populations. Phylogenetic relationships among these haplotypes were inferred from a minimum spanning network, which was constructed by the computer software MINISPNET, and two phylogenetic reconstructions were determined by using maximum likelihood algorithm incorporated in the Phylogenetic Inference Package (PHYLIP) version 3.5c and neighbour joining algorithm incorporated in the software Molecular Evolution Genetic Analysis (MEGA) version 2.0. All these methods exclusively divided the haplotypes into three monophyletic clades corresponding to China Mainland, Northern Japan, and Southern Japan populations respectively. In these populations,the China Mainland population and the Southern Japan population have a relatively closer affinity than either of them with the Northern Japan population. This indirectly supported the hypothesis that ancestral sika deer migrated into Japan islands across at least two land bridges.3. Four hundred and fifty base pairs (bp) of the mitochondrial control region from six sika deers (C. n. taiouanus) collected from Taiwan Province were sequenced. Only one variable site was determined and two haplotypes were identified. These sequences were combined with those previously obtained for sika deers in China Mainland and Japan. AMOVA analysis showed a significant genetic structure (Ost=0.72, Fst=0.69, P<0.001) between Taiwan and Mainland sika deers. The phylogenetic reconstruction and sequence variation analysis both supported that Taiwan sika deer has a closer affinity with China Mainland population than with Japan populations. The origin and evolutionary history of Taiwan sika deers was also addressed.4. Genomic DNA was extracted from some animal products provided by Nanjing Customs. Partial mitochrondrial DNA cytochrome b (cyt b) gene was amplified and sequenced. The original animals of these products were successfully determined by BLAST search of these sequences in GenBank. The present study provided a novel approach for the identification of imported and exported animal products in customs.