The taxonomic status of Papilio maackii and P.syfanius has long been disputed.We conducted a molecular phylogenetic study to evaluate the taxonomic status of P.maackii and P.syfanius.A total of twenty-four P.maackii individuals from six localities and sixteen P.syfanius individuals from two localities were analyzed.We sequenced the partial region of the CO-I gene（about 579 bp） and partial CO-II gene sequence（about 655bp） of the two species.The Kimura-2-Parameter distances among P.maackii and P.syfanius ranged from 0 to 0.6%.Fifteen haplotypes were obtained based on the combined data set.The results strongly supported that all P.maackii individuals and all P.syfanius individuals formed a large clade,and could not be divided into separated clades.This research indicated that the two species have only very recently undergone speciation.
The complete sequences of the mitochondrial DNA genomes of Panthera tigris,Panthera pardus,and Panthera uncia were determined using the polymerase chain reaction method.The lengths of the complete mitochondrial DNA sequences of the three species were 16990,16964,and 16773 bp,respectively.Each of the three mitochondrial DNA genomes included 13 protein-coding genes,22 tRNA,two rRNA,one O L R,and one control region.The structures of the genomes were highly similar to those of Felis catus,Acinonyx jubatus,and Neofelis nebulosa.The phylogenies of the genus Panthera were inferred from two combined mitochondrial sequence data sets and the complete mitochondrial genome sequences,by MP (maximum parsimony),ML (maximum likelihood),and Bayesian analysis.The results showed that Panthera was composed of Panthera leo,P.uncia,P.pardus,Panthera onca,P.tigris,and N.nebulosa,which was included as the most basal member.The phylogeny within Panthera genus was N.nebulosa (P.tigris (P.onca (P.pardus,(P.leo,P.uncia)))).The divergence times for Panthera genus were estimated based on the ML branch lengths and four well-established calibration points.The results showed that at about 11.3 MYA,the Panthera genus separated from other felid species and then evolved into the several species of the genus.In detail,N.nebulosa was estimated to be founded about 8.66 MYA,P.tigris about 6.55 MYA,P.uncia about 4.63 MYA,and P.pardus about 4.35 MYA.All these estimated times were older than those estimated from the fossil records.The divergence event,evolutionary process,speciation,and distribution pattern of P.uncia,a species endemic to the central Asia with core habitats on the Qinghai-Tibetan Plateau and surrounding highlands,mostly correlated with the geological tectonic events and intensive climate shifts that happened at 8,3.6,2.5,and 1.7 MYA on the plateau during the late Cenozoic period.
