| Teinopalpus aureus,belonging to the genus Teinopalpus,is China National butterfly,and a kind of rare and valuable species.It was listed as the First Class Protected Anima by China in 1989 and Date Deficient species in the International Union for Conservation of Nature(IUCN)red list in 2004.There are 5 geographic subspecies documented:Teinopalpus aureus aureus Mell,T.aureus wuyiensis Lee,T.aureus hainanensis Lee,T.aureus guangxiensis Chou et Zhou and T.aureus skinkaii Mori before this study.They distribute in Southeast Asia,north to the Chimu Mountain,Zhejiang Province of China,south to Dong Nai Province of Vietnam.In China,it was found in Zhejiang Province,Jiangxi Province,Fujian Province,Hunan Province,Guangdong Province,Guangxi Zhuang Autonomous Region and Hainan Province.As its unique behavior and habitat,it is rarely seen and difficultly studied.In this study,the complete mitochondrial genome of Teinopalpus aureus was amplified by the conventional PCR,based on preliminary studies of Feng Qin and Ying Huang,who worked in our laboratory from 2008 to 2014.Meanwhile,the primary mitochondrial genomes sequence was revised and the structure and characteristics of the genomes were analysised.According to the mitochondrial rDNA,A-T rich regions and protein coding gene(PCG),MEGA4.2 was used to analysis conserved sites,parsimony informative sites,variable sites transversional pairs(si),transversional pairs(sv),si/sv,genetic distances of Teinopalpus aureus among different geographical populations.To provide a scientific basis for protection zones to formulate measures for the protection of Teinopalpus aureus and strengthen cooperation with each other,the Neighbor-joining phylogenetic tree and the Maximum Parsinomy tree were constructed to investigate phylogenetic relationships among different geographical populations,using Teinopalpus imperialis as the outgroup.The results are as follow:1.The complete mitochondrial genome sequences of T.aureus is 15234 bp to 15239 bp in length,and contains 13 PCGs(ATP6,ATP8,Cyt b,COI-?,ND1-6,ND4L),2 rDNAs(12S rRNA,16S rRNA),22 tRNA genes and 1 A+T-rich region.The order of the sequences and transciption protein-coding genes are the same as those that had reported,without any gene rearrangement or missing.There are 12 spacer sequences from 1 bp to47 bp in length,with a largest spacer sequences between gene tRNAGln and ND2.The spacer sequences between tRNAAsn and tRNASer(AGN)in DYS1 is 6 bp(TTTATA),and the others are 5 bp(TTATA).There are 11 or 12 overlapping sequences.The region between tRNAAla and tRNAArg is no overlap area in DYS1,but there is an overlap of 2bp in DYS2.While in the region are an overlap of 2bp in the rest of the individual.At the same time,the region between Cytb and tRNASer(UCN)is no overlap area in HN1.While in the region are an overlap of 2bp in the other individuals.The A+T content of the mitochodrinal genome is79.8%,having the same feature of AT bias within each gene.Other genes use the initiation codons "ATN",except for COI initiation with CGN;Gene COI,COII,ND3 and ND4 terminate into a single T,while others use common stop codons "TAN".All the tRNA genes have the typical leaf clover secondary structures,except for the tRNASer(AGN),which lacks the DHU arm.In different individuals,tRNA length and the base composition have some difference which individual nucleotide mutated on D-Loop,T?C Loop and Variable Loop,but this do not affect tRNA function normally.The length of 16S rRNA and 12S rRNA is different in different individuals.The 12S rRNA gene is from776 bp to777 bp in length among different geographical populations,while the 16S rRNA gene is from 1334 bp to 1336 bp.The A+T content of 16S rRNA and 12S rRNA were 82.1%and 85.8%,respectively.The A+T region is 401bp in length,and the A+T content is 93.4%in this region.There are some tandem repetitive sequence:(AT)6,(AT)9G(TA)4.In addition,there are two repeat sequences"AAATTAATAAATTA".The 6bp sequence near the tRNAMet gene are different in different individuals.There are "AAATAA" in HN1 and HN2,while in the other individual are poly-A typical 6 bp sequence.2.Based on phylogenetic analysis of rDNA,A-T rich regions and protein coding gene of different geographic populations of Teinopalpus aureus,the results showed that the genetic distances among Dayaoshan population,Wuyishan population and Nanling population were less than that between them with Hainan population and Vietnam population,and the genetic distance between Hainan population and Vietnam population was relatively large.In the NJ tree and MP tree,Dayaoshan population,Wuyishan population and the Nanling population alone did not constituted a separate lineage,but as a clade.At the same time,Hainan population and Vietnam population were almost not clustered in a clade with the population that shown above.So based on the above data,the results of this study suggested that Teinopalpus aureus should reasonably divide into Teinopalpus aureus aureus(including Teinopalpus aureus aureus,T.aureus wuyiensis,T.aureus guangxiensis),T.aureus hainanensis and T.aureus skinkaii.According to Chinese glacial evolution and genetic diversity of different populations,this study inferred Nanling region was most likely to be the the origin of Teinopalpus aureus. |
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