Mitochondrial Genome Sequenceing Of Four Cynipidae And Phylogenetic Relationship Analyzing Among Proctotrupomopha

Cynipidae belongs to the Hymenoptera,is the only pests for plants and forests.The technology of the mitochondrial DNA sequence gives the possibility for futher research of insect phylogenetic relationship in various level.Currently the database of NCBI totally has 113 mitochondrial genomes of Hymenoptera insect records.The records mainly focus on the large individuals,including species of Braconidae,Apoidae,Ichnenmonoidea and Tenthredinoidea eg.Only one species of Cynipoidea Ibalia lencospoides(KJ 814197)has acquires the complete mitochondrial genome sequence.The classification and internal relationship among the Proctotrupomorpha is still elusive and controversial.In this study,the method of Next-generation sequence and platform Illumina Hiseq are used to sequence the whole mitochondrial sequences of four Cynipidae species: Andricus flavus,Andricus sp.nov.(Andricus),Synergus castaneus(Synergus),Dryocosmus kuriphilus(Dryocomus),also with the splicing,annotating,composition analyzing.Combining with the comparative genomics and Bioinformatic,the arrangement of the four species is compared with the ancestral insect.The discussion of evolutionary selection pressure through the calculating combining the data set of protein coding genes(PCGs)of 12 proctotrupomorpha species have been included in the NCBI,with Diadromus(Ichneumonoidea)and Spathius(Braconidae)as outgroop,using Maximum likelihood and Bayesian method construct the phylogenetic tree and analysis the phylogentic relationship.Main coclusions of the research are summarized as following:1.The sequence length of Andricus falvus,Andricus sp.nov.,Synergus Castaneus and Dryocosmus kuriphilus are 18043 bp,19035bp,19167 bp and 14005 bp respectively.Andricus falvus encodes 12 protein genes,10 t RNA genes;Synergus Castaneus encodes 13 protein genes,2 r RNA genes,21 t RNA genes;Dryocosmus kuriphilus encodes 12 protein genes,1 r RNA genes,16 t RNA genes.2.The whole mt DNA and the different parts base compositions of four materials showed A+T bias,A+T contentis greater than the G+C content.Compare the AT skew and GC skew value,different bases were used in different sites.The third positions of all Cynipidae have the highest A+T percent.The shart coclon usage of Andricus falvus,Andricus sp.nov.,Synergus Castaneus and Dryocosmus kuriphilus.mitochondriak PCGs are all ATN,including ATA?ATT?ATC?ATG.Majority PCGs stop codon usage are TAA,while several use incomplete termination codon TA or T.Majority tRNA genes of four species mitochondrial genomes present a typical clover leaf structure except trn A,trn H,trn S1 Dryocosmus kuriphilus,with lack of arms.Some mistaches exist in t RNAs and maily the G-U mismatched base pair.The most frequently used condon is uu A(leu1),(The relative synonymous Codon usage)greater than 1 in the condon third site with the base A or U.3.The arrangement of the Cynipidae was vary from ancestral insect,several genes have rearranged in position and direction.4.By calculating the Ka,Ks,and Ka/Ks values of the four wasp protein coding genes,it was found that the insects of the Cynipidae were purified during evolution.5.Phylogenetic relationships of Proctotrupomorpha insects were constructed with 12 species all protein coding genes through ML and BI respectively.The relationship Proctotrupomorpha in general level is elusive,while in section level it is stable.