Yunnan Card Locust, Spotted Legs Mengla Locust Coriolis Winged Locust Mitochondrial Genome Sequencing And Analysis

For a long time,Orthopterists have been researching phylogenetic relationships among groups of the Orthoptera.Although Ensifera and Caelifera are accepted as monophyletic taxa,some Orthopterists believe that they have different taxonomic ranks.Animal mitochondrial genomes are used as a useful model for structural,functional and evolutionary studies.Complete mitochondrial genome sequences were available in GenBank for 1537 metazoan species,of which 116 for Insecta.In this paper,the complete mitochondrial genomes of Carsula yunnana,Menglacris maculata and Zubovskia koeppeni were sequenced,assemblyed,annotated and analyzed in details with the strategies of long-PCR and sub-PCR.The three new mitochondrial genomes combined with that of other insects deposited in GenBank,as well as other Orthoptera species which completed mitochondrial genome were sequenced in our laboratory were used to reconstructed Orthoptera phylogeny and explored the phylogenetic utility of different combined data set.The main conclusions are summarized as following.1.Carsula yunnana,Menglacris maculata and Zubovskia koeppeni are 15 444 bp,15 602 bp and 15815 bp respectively.They all encode 13 protein genes:COâ… ,COâ…¡,COâ…¢,ATP6 and ATP8;ND1,ND2,ND3,ND4,ND4L,ND5,ND6å'ŒCytb,2 rRNA genes:1rRNA and srRNA, 22 tRNA genes and A+T rich region。2.The average A+T content of Carsula yunnana,Menglacris maculata and Zubovskia koeppeni mitochondrial genome has strongly the A+T bias.The strongest A+T biase is found in Carsula yunnana which is nearly 76.1%of the three species,the weakest found in Menglacris maculata which is 74%3.COâ… and ND5 genes in the Carsula yunnana,Menglacris maculata and Zubovskia koeppeni had abnormal start and stop codons.The putative initiation codon for cox1 appears to be ACC in the Menglacris maculate,ND5 gene has incomplete termination codon(TA and T) in the Carsula yunnana and Zubovskia koeppeni mitogenome respectively.This phenomenon is popular in the other Orthoptera mitogenome.4.All the 22 tRNA genes of Carsula yunnana,Menglacris maculata and Zubovskia koeppeni mitochondrial genomes have a typical clover leaf structure except tRNA Ser(AGN),which is lack the DHU arm.5.The complete secondary structures of the rRNA genes in Carsula yunnana,Menglacris maculata and Zubovskia koeppeni mitogenome are predicted.Most parts of the secondary structure among different species are similar,the main differences appear in two terminal regions. 6.The A+T contents of the A+T-rich regions of the 22 Orthoptera insects vary from 66.8% (Troglophilus neglectus) to 87.3%(Acrida willemsei).We found that Acridoidea insects generally have the conserved block,such as Block A,Block B,Block E1 and E2,of which Block E1 and E2 can form certain secondary structure which connects with the origion replication of the N chain.But Ensifera insects do not find this block obviously.7.Phylogenetic analyses from the protein coding gene sequences support two suborder Caelifera and Ensifera as monophyletic groups.Tettigonioidea was supported as monophyletic groups too.Within Acridoidea,Tridactylus japonicus was the acncestral group.Pyrgomorphidae and Chrotogonidae have closely relationships,Oedipodidae,showed the monophyly,but Catantopidae, Acrididae,and Arcypteridae were not accepted as monophyletic groups respectively.