DNA Barcoding And Phylogeny Research On Some Species Of Phasmatidae From China

Phasmatidae is the larger family of Phasmida from insecta,and about 8 subfamilies,21 tribes,128 genera,674 species and 28 subspecies are known in the world,and 3 subfamilies,6 tribes,22 genera,189 species and 1 subspecies have been reported in China,among them,there are both economic insects and forestry pests.Due to their development is incomplete metamorphosis and strong androgyny exist,lead to the traditional morphological taxonomy is difficult.Therefore this study was conducted through the classification of some genera and species of Phasmatidae in China,while extracting their total DNA,and amplifying the 28 S ribosome DNA,Mitochondrial cytochrome Oxidase subunit I(COI)and Mitochondrial cytochrome Oxidase subunit II(COII)in order to achieve rapid identification,prevention and research.In this way,the DNA barcoding database were improved and provide an assistance for the Phasmatidae classification and identification.The phylogenetic tree was also constructed based on the obtained high-quality sequences through the following four methods by Bayesian inference(BI),Maximum likelihood(ML),Maximum parsimony(MP)and Neighbor joining(NJ).(1)The results of the morphological classification research showed that 3 subfamilies,6 tribes,24 genera,202 species,2 subspecies were recorded in this paper,and the keys of the corresponding subfamilies,genera and species were compiled.also described 2 new genera,12 new species and 1 new subspecies,they were Sinoacrophylla Xu & Jiang gen.nov.,Ramtirachoidea Xu & Jiang gen.nov.,Sinoacrophylla longtanensis Xu & Jiang sp.nov.,Sinoacrophylla banzhaiensis Xu & Jiang sp.nov.,Ramtirachoidea hainanensis Xu & Jiang sp.nov.,Entoria parahei Xu & Jiang sp.nov.,Entoria inermus Xu & Jiang sp.nov.,Ramulus chinensis Xu & Jiang sp.nov.,Ramulus sparsiserrulate Xu & Jiang sp.nov.,Ramulus exilisianalus Xu & Jiang gen.nov.,Ramulus albopunctatus Xu & Jiang sp.nov.,Ramulus longiantenna Xu & Jiang sp.nov.,Cnipsomorpha jinpingensis Xu & Jiang sp.nov.,Cnipsomorpha yuxiensis Xu & Jiang sp.nov.,Phryganistria guangxiensis longtanensis Xu& Yang subsp.nov.Supplementary described of the characteristics of the opposite sex of the three species:male of Ramulus bomiense(Chen & He,1994)and Ramulus mikado(Rehn,1904),female of Ramulus fasciatus(Chen & He,2006).In addition,the Chinese names of 1 family,4 genera and 25 species are first recorded.(2)This research involved more than 300 specimens(collected by the research group,including duplicate specimens,which cannot be distinguished from specimens,and Specimens from other families),and DNA extraction was performed on 54 specimens from the Phasmatidae,involved 39 species of 11 genera.A COII primer and 95 DNA barcodes were obtained,included 34 28 S barcodes,related to 7 genera 34 species,25 COI barcodes,related to 5 genera 25 species,36 COII barcodes,related to 11 genera 36 species.(for detailed information,please refer to Table 1,3 for Genkank number).(3)Furthermore research through phylogenetic study showed that r DNA could be constructed a more reliable phylogenetic tree by BI method,while mt DNA by NJ method or ML method,At the same time,we have obtained more reliable phylogenetic relationships of some species of Chinese Phasmatidae:Subfamily: Platycraninae +(Phasmatinae + Clitumninae)?Genus: Platycranini +((Stephanacridini + Phasmatini)+(Pharnaciini +(Medaurini +Clitumnini)))?Tribes: Megacrania +((Phasmotaenia + Sinoacrophylla)+((Ramtirachoidea+(Phryganistria + Tirachoidea))+(((Cnipsomorpha + Parapachymorpha)+(Neo Cnipsomorpha +(Interphasma + Neosinophasma))+(Ramulus +(Entoria +Rhamphophasma)))))?...