Determination And Analysis Of The Whole Mitochondrial Genome Of Six Species Of Longhorn Beetle

The mitogenome is becoming increasingly used in many reserach fields,such as population genetics,species identification and phylogenetic studies due to its stable composition,maternal inheritance,containing a large number of molecular markers.Cerambycidae belongs to coleoptera chrysomeloidea,is one of the most diverse groups in coleoptera,as well as the main pest harm the forestry production,crop cultivation and construction wood.Compare to other insects,cerambycidae have sequenced 13 complete mitogenomes and there are over 25000 species in Cerambycidae.Therefore,this study select six important forestry pests of cerambycidae,hoping to increase the richness of the cerambycidae genome information.At the same time,this study can provide some effctive information in phylogenetic.In this study we take the method of Illumina and tranditional sanger to sequence,comment and analyze the complete mitogenome of Ommata notabilis,Stenygrinum quadrinotatum,Glenea centroguttata,Arhopalus rusticus,Prionus insularis,Thraninus simplex.The results are followed;1.The length of the above cerambycidae mitogenomes is 15715bp,16091 bp,15561bp,15801bp,15641bp in size,respectively.All of them contain 37 genes and a non-coding region.There aren't gene rearrangement.2.The six mitogenomes and all of the compositions show aboviously high AT biase.In addition,the complete mitogenomes of six cerambycidae show A biase and C biase,but the most components of the mitogenomes show T biase and G biase.3.The main start codons in protein coding genes of six mitogenomes are typical ATN.The start codon of ND1 in Ommata notabilis,Stenygrinum quadrinotatum,Glenea centroguttata,Arhopalus rusticus is TTG.In Prionus insularis,the start codon is GTA,the stop codon is CCG.Among the avaliable codons,the most frequently used codons are UUU,UUA,AUU,AUA which highly contain A and U.The higher amino acid composition are Leu(Leucine),Ile(isoleucine),Ser(serine),Phe(phenylalanine).4.In the secondary structure of the 22tRNAs,21 tRNAs form a typical clover leaf structure,except tRNAser(AGN),which lack of a DHU stem.Unlike other arthropods,the tRNAser(AGN)use UCU instead of GCU as anticodon.There are a few mismatches of the base,G-U and is the main type.A-G mismatch was found in the tRNATrp,U-U mismatch was found in tRNALeu(UUR).5.In this study,we amplified COI and AT-region by the First-generation Sequencing Technology.The match rate of the two regions between the First-generation Sequencing Technology and Next-generation Sequencing Technology is 99.5%-100%.6.The comparative mitogenomics analysis in 28 species found the mitogenomes are about 1.54?1.66kb.The larger overlapping regions are in tRNATrp-tRNACys,COI-tRNATyr,the larger intergenic region is between ND1 and tRNASer.The A+T content of every component in the mitogenomes of 28 species is about 70%and the bias is A>T.The largest change of gene is ND3,the smallest is COIII.The evolution rate of ATP8 gene is fastest and COI gene is lowest.Most of the protein-coding genes use typical ATN as the start codon and TAA,TA,T as the stop codon.7.The sequence composition,genetic distance,sequence heterogeneity,basepair substitution of 11 PCGs in 28 species were analyzed,and the construction of phylogenetic tree was carried out.The highest percentages of the conserved site in protein coding gene is COI.The biggest genetic distance is between Batocera lineolata and Apriona swainsoni and the smallest is between Disteniinae.sp and obrium sp.The basepair substitution and sequence heterogeneity analysis show the 28 species nucleotide datasets can be used for construction of phylogenetic tree.The result of phylogenetic tree obtain the supports from comparative anatomy and biogeography.Two methods(ML and BI)are basically found the same results.This study established the six kinds of cerambycidae basic position in phylogenetic tree and the relationship among subfamilies of cerambycidae.The subfamilies of cerambycidae relationship were(Lamiinae+((Aseminae+(Necydaline+Lepturinae))+((Vesperidae+Disteniinae)+(Prioninae+Cerambycinae)))).Our results support the monophyly of the Lepturinae,Lamiinae,Cerambycinae and Batocerini,Saperdini,Lamiini,Clytini.