Systematic Classification Of Difficult Groups In The Tribe Olethreutini From China(Insecta:Lepidoptera:Tortricidae)

Olethreutini walsingham,with over 1200 described species,is one of the most diverse tribes in the family Tortricidae.However,it is not confirmed that this tribe is a monophyletic group,and the classification has been debated as to whether the four groups,Microcorsini,Gatesclarkeanae,Bactrae and Endotheniae,are members of Olethreutini.To define the monophyletic Olethreutini and clarify the taxonomic positions of the difficult groups,we recorded Chinese species of the four difficult groups,sequenced their COI genes and mitochondrial genomes,and presented the molecular estimate of relationships across the subtribes of Olethreutini and the tribes of Olethreutinae.The results are as followed.1.47 species in 7 genera from China are recognized.There are four species described as new:Syntozyga xizangica sp.nov.,Endothenia asymmetrica sp.nov.,E.bicruris sp.nov.and E.distensus sp.nov..One genus and eleven species are newly recorded for China: Ukamenia Oku,1980,U.sapporensis Mastsumura,1931,Cryptaspasma angulicostana Walsingham,1900,C.mirabilis Kuznetzov,1964,Bactra robustana(Christoph,1872),Syntozyga cerchnograpta Razowski,2014,S.pedias(Meyrick,1920),Endothenia designata Kuznetzov,1970,E.bira Kawabe,1976,E.menthivora Oku,1963,E.polymetalla(Turner,1916)and E.villosula Falkovtsh,1966.2.63 effective sequences(658bp)of COI gene were obtained,which from 19 species of 7genera.Among them,4 sequences under 2 species of Cryptaspasma,2 sequences under 1specie of Gatesclarkeana,2 sequences under 1 specie of Hiroshiinoueana,1 sequence from 1specie of Ukamenia,21 sequences belong to 5 species of Bactra,6 sequences under 2 species of Syntozyga,27 sequences under 7 species of Endothenia.Combined with 325 effective sequences reported in NCBI database(27 sequences from 12 species of Cryptaspasma,7sequences from 4 species of Gatesclarkeana,3 sequences from 2 species of Hiroshiinoueana,120 sequences from 16 species of Bactra,7 sequences from 3 species of Syntozyga and 161 sequences under 16 species of Endothenia),we did the analysis of the genetic distance between species and within species.The average genetic distance between the species of Cryptaspasma is from 4.56% to 14.23%,with an average value of 11.83%,the average intraspecies genetic distance is between 0 and 3.84%,with an average value of 1.18%;the average genetic distance between the species of Gatesclarkeana is from 4.72% to 15.72%,with an average value of 8.93%;the average genetic distance between the species of Hiroshiinoueana is 8.86%,the average intraspecies genetic distance is 0.97%;the genetic distance of Bactra species is ranging from 2.20% to 13.20%,with an average value of 7.98%,the average intraspecies genetic distance is between 0~1.59%,with an average value of 0.48%;the genetic distance between the species of Syntozyga is from 2.28% to 8.36%,and the average is 6.46%;the genetic distance between the species of Endothenia is from 2.35% to12.59%,with an average value of 7.84%,the average intraspecies genetic distance is between0~2.51%,with an average value of 0.58%.According to the DNA Barcodes(COI),we verified the identity of Endothenia genitanaeana.3.14 complete mitochondrial genomes were sequencinged,including 6 genomes from the difficult groups,6 genomes belong to the core groups of Olethreutini,1 genome from Enarmoniini and 1 genome from the Cochylini of Tortricinae.37 genes of sequences from the difficult groups have the same arrangement order and direction with those recorded in Lepidoptera.Compared with the C+G base content,the A+T base content makes up a larger proportion.Except COI gene,other PCGs use ATN as start codons and all of PCGs end with TAA or T-t RNA.7 complete mitochondrial genome sequences were downloaded in NCBI(including 2 genera of Eucosmini,2 genera of Grapholitini and 3 genera of Tortricinae).Combined with all the complete mitochondrial genome data,a ML tree was constructed for phylogenetic analysis.The results showed that Gatesclarkeanae,Bactrae and Endotheniae are closely related to each other,and they constitute a monophyletic group with the core group of Olethreutini.In conclusion,the Olethreutini should include these three taxa and become a natural taxon.Our results are consistent with Diaknoff' s opinion(1973)of composition of Olethreutini.And it roughly the same with the view of Regier(2012)based on the results of the nuclear genes.It shows that the Microcorsini is far from the core group of Olethreutini,and it is an early branch in the Olethreutinae and should be a single branch.