Taxonomy And Phylogeny Of The Family Caracidae In Chin

The planthopper family Caliscelidae(Hemiptera: Auchenorrhyncha: Fulgoroidea)is a small group of superfamily Fulgoroidea.With various body types,it can be divided into long-winged type and short-winged type.Worldwide distribution.By the end of 2019,two subfamilies,five tribes,78 genera,and 245 species had been recorded in the world,and two subfamilies,five tribes,15 genera,and 38 species had been recorded in China.Caliscelidae are all herbivorous.Many groups specialize in feeding on bamboo,and many of them are important or potential pests on bamboo.At present,the main problems in the classification of Caliscelidae are as follows: the species diversity of Caliscelidae in China is not well studied,and there are still many new taxa to be discovered and described;the phylogenetic status of the Caliscelidae in the Fulgoroidea is unclear;the relationship among the higher order members of Caliscelidae is questionable;at the same time,previous phylogenetic research lacks mitochondrial genome data to explore the problem.The present dissertation is a systematic study on the family Caliscelidae from China.The main contents include: taxonomic research,description and comparative analysis of the mitochondrial genomes of representative species,phylogenetic studies based on morphological characteristics,phylogenetic evolution of Caliscelidae based on mitochondrial genome data,and reconstruction of phylogenetic relationship based on mitochondrial genome data.The results are as follows:1.TaxonomyIn this paper,a comprehensive and systematic taxonomy of family Caliscelidae from China has been carried out.There were two subfamilies,five tribes,23 genera and 57 species of family Caliscelidae recorded.More than 2000 specimens were examined,two subfamilies,four tribes,18 genera and 41 species,included four new genera and 17 new species were identified,and four genera and two species were recorded from China for the first time.The main morphological characteristic figures were provided.The identification characteristics and keys to subfamilies,tribes,genera and species were also provided respectively.Four new genera were established: Tuberculum gen.nov.;Fuscinulum gen.nov.;Neosymplana Chen & Gong,2020(Previously published);Parasymplanella gen.nov..Seventeen new species were discovered: Bambusicaliscelis bibulbus sp.nov.;Bambusicaliscelis clavatus sp.nov.;Bambusicaliscelis anlongensisi sp.nov.;Bambusicaliscelis nigritus sp.nov.;Bambusicaliscelis suiyangensis sp.nov.;Tuberculum maculata sp.nov.;Annamatissus mohanium sp.nov.;Peltonotellus lobulus sp.nov.;Peltonotellus albulus sp.nov.;Fuscinulum rubellus sp.nov.;Symplanella fulva Gong & Chen,2020(Previously published);Symplanella nigricans Gong & Chen,2020(Previously published);Neosymplana vittatum Gong,Yang & Chen,2020(Previously published);Augilina triaina Chen & Gong,2021(Previously published);Augilina tetraina Chen & Gong,2021(Previously published);Anthracidium maculatus sp.nov;Parasymplanella maximua sp.nov..Four genera and two species were recorded from China for the first time.New record genera: Chirodisca Emeljanov,1996(Previously distributed in Africa,etc.);Anthracidium Emeljanov,2013(Previously distributed in Vietnam);Annamatissus Gnezdilov & Bourgoin,2014(Previously distributed in Vietnam);Augilina Melichar,1914(Previously distributed in Vietnam and Philippines);New record species: Chirodisca eximia,(St?l,1859);Annamatissus tami Gnezdilov & Soulier-Perkins,2014.2.Description and comparative analysis of mitochondrial genome of CaliscelidaeTwenty-one complete mitochondrial genome sequences were obtained by total DNA extraction,high-throughput sequencing,comparative splicing and annotation.The description and comparative analysis of 13 protein-coding genes,22 t RNA,2 r RNA and control regions of mitochondrial genome showed that:(1)The mitochondrial genome of 21 species of Caliscelidae were sequenced,representing two subfamilies,three tribes and 15 genera of Caliscelidae in China.The subfamilies,tribes and genera accounted for 100%,60.0% and 69.6%,respectively.The size of the mitogenomes of the Caliscelidae is betwcen 15.2 kb-16.7 kb.The length is mainly determined by the size of the control region,and is also related to the number and size of the non-coding regions.(2)The mitochondrial genome is conserved without gene rearrangement,addition or deletion.There are short spaces or even overlaps between genes in mitochondrial genomes.An eight-bp overlap between trn W and trn C,a seven-bp overlap between ATP8 and ATP6(except Caliscelisshandongensis),and a seven-bp overlap between nd4 and nd4L(except Youtuus erythrus),were found in this study.Overlap is relatively consistent among tribes,genera and species of Caliscelidae.(3)The AT content of the mitogenomes was significantly higher than the GC content,showing an AT bias.The 13 protein-coding genes often use the typical ATN as the start codon.GTG is also commonly used as the start codon in nd1 and nd5.The stop codon is often a typical TAN,in a few genes,such as cox1,atp6,etc.,and incomplete stop codon T is often adopted.The 5 amino acids with the highest codon usage among the 22 amino acids in each species were Phe,Ile,Leu2,Met and Ser2.(4)The results of evolution rate analysis indicated that cox1 showed the strongest purification selection,ND-family genes showed slightly loose purification selection,and atp8 showed the smallest selection pressure and the fastest evolution speed.(5)In Caliscelinae,trn S1 and trn V lack the dihydrouridine arm(DHU stem)to form a simple loop;in Ommatidiotinae,trn C,trn S1 and trn V lack the dihydrouridine arm to form a simple loop;while the rest of the genes are folded into a typical clover secondary structure.The secondary structure of t RNA shows correlation among subfamilies.3.Research on evolutionary status of Caliscelidae based on mitochondrial genomeThe two representative species of the family Cicadellidae,which are relatively closely related,were taken as the outgroup,and the 17 families of the superfamily Fulgoroidea were taken as the ingroup.The nucleotide sequences of 13 protein-encoding genes were selected to analyze the evolutionary status of Caliscelidae in the Fulgoroidea by using Bayesian(BI)and Maximum likelihood(ML).The results showed that:(1)The 15 genera of three tribes and two subfamilies of Caliscelidae(representing Peltonotellini,Caliscelini and Augilini)were always clustered into one branch,indicating that the Caliscelidae is a natural monophyletic group.Based on the mitochondrial genome,it is supported to separate this group from the Issidae and become an independent family.(2)Caliscelidae is one of the most evolved groups in the superfamily Fulgoroidea.The most closely related groups of the family are Ricaniidae,Flatidae and Issidae.4.Phylogenetic analysis of genus-level elements based on morphological characteristicsGergithus Quinquemaculatus were used as the outgroup,and 23 genera of five tribes and two subfamilies were used as the ingroup.Based on 84 morphological characters,a phylogenetic study of the genus-level elements of the Chinese Caliscelidae was carried out using the maximum parsimony(MP).The results showed that:(1)It is supported that Caliscelidae can be divided into two subfamilies,the subfamily Caliscelinae includes Peltonotellini and Caliscelini,and the subfamily Ommatidiotinae includes Adenissini,Ommatidiotini and Augilini.It is supported to divide Caliscelidae into five tribes,and the phylogenetic relationship among the five tribes is:((Peltonotellini + Caliscelini)+(Adenissini +(Ommatidiotini + Augilini))),Augilini was the latest in differentiation and was a more evolved group.(2)The tribe Caliscelini currently consists of eight genera and their phylogenetic relationships are as follows:((Chirodisca+(((Gelastissus + Nenasa)+ Annamatissus)+ Caliscelis)+(Tuberculum+ Cylindratus+Bambusicaliscelis))).(3)The tribe Adenissini currently includes three genera,and the phylogenetic relationship among them can be shown as:(Phusta+(Bocra+Delhina)).(4)The tribe Augilini currently includes 10 genera and their phylogenetic relationships are:(Youtuus +(Anthracidium +((Fuscinulum + Augilodes)+((Augilina + Symplana))+((Neosymplana +(Parasymplanella + Pseudosymplanella))+ Symplanella)))).5.Phylogenetic analysis of genus-level based on mitochondrial genome among the CaliscelidaeThe five representative species of the closely related Issidae,Ricaniidae and Flatidae were taken as the outgroup,and two subfamilies three tribes and 15 genera of the Caliscelidae were selected as the ingroup.The nucleotide sequences of 13 protein-coding genes were selected,using Bayesian(BI)and Maximum likelihood(ML)to perform phylogenetic analysis of the genus-level elements of the family Caliscelidae in China.The results showed that:(1)It is supported that Caliscelidae can be divided into two subfamilies,namely Caliscelinae and Ommatidiotinae.The subfamily Caliscelinae includes Peltonotellini and Caliscelini,and the subfamily Ommatidiotinae includes Augilini,which are consistent with the results of phylogenetic analysis based on morphological characteristics.(2)The tribe Caliscelini currently consists of four genera and their phylogenetic relationships are asfollows:(Caliscelis +(Tuberculum +(Cylindratus+ Bambusicaliscelis))).(3)The tribe Augilini currently consists of ten genera and the phylogenetic relationships among them are as follows:(Youtuus +(Anthracidium +((Fuscinulum + Augilodes)+((Symplana + Augilina)+((Neosymplana +(Parasymplanella + Pseudosymplanella))+ Symplanella))))).(4)Fuscinulum and Augilodes,Symplana and Augilina,Parasymplanella and Pseudosymplanella are sister groups,which are consistent with the phylogenetic analysis results based on morphological characteristics.