The Mitochondrial Genome And Molecular Phylogenetic Analysis Of Manis Pentadactyla (Mammalia:Pholidota)

The Manis pentadactyla belongs to Manidae which are mainly living in the south areas of the Yangtze River of China, and Nepal, Sikkim and Burma. Since the1980s, due to the ecological environment deteriorate and excessive hunting, the M.pentadactyla population has declined dramatically in china. Therefore, M.pentadactyla is listed as a vulnerable species by International Union for Conservation of Nature and Natural Resources (IUCN) Red List and also as a secondary endanger species in China.The mitochondrial genome of the M.pentadactyla has been sequenced in this paper. To determine the phylogentic position of M.pentadactyla with related species,50representative species of the Mammalia were reconstructed using phylogenetic analysis softwares, the phylogenetic tree was reconstructed with the concatenated nucleotide dataset of the12heavy-strand-encoded protein genes.The phylogenetic analysis was carried out using maximum parsimony (MP) and Bayesian inference (BI) methods. Meanwhile, mitochondrial genes (12rRNAand16rRNA) were used as molecular markers, using Tachyglossus aculeatus and Ornithorhynchus anatinus as the outgroups, the phylogenetic trees were reconstructed with Neighbor-joining (NJ) and Bayesian Inference (BI) methods. After analysing and discussing these results, we draw the conclusions as follows:1. The complete mitochondrial genome of M.pentadactyla was sequenced and analyzed using overlapping long PCR amplification. The mtDNA of M.pentadactyla was16,578bp in length, contained13protein-coding genes (ATP6, ATP8, COI-III ND1-6, ND4L, Cytb),2ribosomal RNAs (12S rRNA and16S rRNA),22transfer RNAs and a putative control region (D-loop).Its gene arrangement pattern and orientation were the same with that of other species of Mammalia.The complete mitochondrial sequences of30species from9orders of Mammal were obtained from GenBank.The maximum-parsimony (MP) methods and bayesian inference (BI) methods were reconstructed based on the concatenated nucleotide sequences of12protein-coding genes.The results support strongly the M.pentadactyla and Carnivora is sister branch.2. Results of phylogenetic analysis based on partial sequence of mitochondrial12S rRNA gene showed that50species from9orders clustered into three clades.The first clade includes those of the orders Carnivora, Artiodactyla, Rodentia, Insectibora and Pholidota,4orders are separated from each other in the molecular phylogenetic trees.In the first major branch, Carnivora and Artiodactyla clustered into the first small branch, Rodentia and Insectibora clustered into a form sister branch, Pholidota formed an independent branches.The second one includes those of the orders Chiroptera and Primates.The third clade includes the orders Peramelemorphia and Diprotodontia.3. The BI and NJ molecular phylogenetic trees based on16S rRNA gene indicated that3Superorder and9order are separated from each other in the molecular phylogenetic trees.The first clade includes those of order Carnivora, Artiodactyla,Chiroptera and Pholidota from Superorder Laurasiatheria. In the first major branch, Carnivora and Artiodactyla clustered into the first small branch, than meeting with Chiroptera. The second one includes those of the orders Peramelemorphia and Diprotodontia from Australidelphia.The other species belonging to Primata, Lagomorpha and Primata formed sister branch, and Rodentia formed an independent branches in the branch.The result is similar to the classification based on traditional morphology.