Taxonomy And DNA Barcoding Of Anling Wax Cicadas In China

The planthopper tribe Andini(Hemiptera: Auchenorrhyncha: Fulgoromorpha: Cixiidae:Cixiinae)consists of 123 species in 3 genera in the world and 13 species in 2 genera in China.As a phytophagous insect,the tribe Andini is a potential threat to agricultural and forestry production.At present,the research of species diversity of the tribe Andini in China is weak and unclear,and the appearance and body-color markings among the individuals are very similar.Therefore,molecular data should be combined to accurately identify the taxa.In order to grasp the species composition of the tribe Andini furtherly,find more new taxa,enrich the species resource bank,build a molecular database of this tribe,and solve problems such as the identification of related and difficult species.In this paper,a systematic taxonomic study of the tribe Andini in China is carried out.The main contents include morphological classification,DNA barcode research,and identification of doubtful species based on DNA barcode.The main findings are as follows:Taxonomy1.A systematic taxonomic study of the tribe Andini in China is carried out.31 species of 3 genera of Andini are recorded in China,among them,including 19 new species,1 new record genera and 1 new combination.The male genitalia of 23 species of Andini in China were mapped and the color pictures of adults were provided.Based on this,the keys to the genera and species of Andini in China was compiled.The type specimens and examined specimens were deposited in the Institute of Entomology,Guizhou University,Guiyang,China(GUGC).2.19 new species,1 newly recorded species from China,and 1 new combination were found.The new species discovered,newly recorded in China and new combination species proposed are listed as follows:New species(19 species):Andes curvispinus sp.nov.Andes dentatus sp.nov.Andes furcutus sp.nov.Andes fuscstictus sp.nov.Andes latanalus sp.nov.Andes melanohabenus sp.nov.Andes longiflagelis sp.nov.Andes pallidus sp.nov.Andixius cultratus Wang,Zhi & Chen,2020(Published)Andixius lingulatus Wang,Zhi & Chen,2020(Published)Andixius bicoloratus sp.nov.Andixius crispiprocessus sp.nov.Andixius motuoensis sp.nov.Andixius flagellihamus sp.nov.Andixius gracilispinus sp.nov.Andixius productus sp.nov.Parandes apicalis sp.nov.Parandes circinatus sp.nov.Parandes lobatus sp.nov.New records species from China(1 species):Parandes hemina(Fennah,1978)New combination(1 species):Parandes hemina(Fennah,1978),comb.nov.(from Andes St(?)l,1866)Study on DNA barcode1.The tribe Andini DNA barcode candidate molecular markers were initially screened.Among the sequence component analysis,R value and base substitution analysis,genetic distance and phylogenetic signal monitoring,the mitochondrial genes COI,COII,Cytb and 16 S rDNA were consistent with the mitochondrial base composition of insects and could provide phylogenetic information.16 S rDNA and Cytb gene sequences had higher variable sites and lower conserved sites,with the R values of 0.60 and 0.62,respectively,indicating unbefitting interspecific genetic distance.The variable sites and conservation sites of COI and COII gene sequences were high,and the R values were 1.04 and 0.85,respectively,indicating moderate interspecific genetic distance.Therefore,COI and COII gene sequences can be used as candidate molecular markers for interspecific species identification of the tribe Andini.2.The DNA barcode database of the tribe Andini was preliminarily constructed.Total DNA was extracted and two gene fragments of COI and COII were sequenced in 13 species of Andini belonging to 3 genera,and 26 available sequences were obtained.It includs 13 species of COI and 13 species of COII,which initially made up the molecular database of the tribe Andini.Based on COI gene sequence,the genetic distances between species and genera were0.1224-0.2069 and 0.1435-0.3784,respectively.Based on COII gene sequence,the genetic distances between species and genera were 0.1785-0.3037 and 0.2445-0.6464,respectively.Identification of doubtful species based on DNA barcodeBased on COI and COII gene sequences,adopted the genetic distance method and cladistic analysis were used to identify the suspected species of the tribe Andini.The results showed that the interspecific genetic distance among Andes hemina and the other five species of the genera of Andes was greater than that among A.hemina and the species of the genera of Parandes.In the phylogenetic tree,A.hemina was gathered to the genus of Parandes.Based on the results of DNA barcode,it is suggested that A.hemina should be transplanted to the genus of Parandes as a new combination.