Cloud Spots Car Locust Mitochondrial Genome Sequencing And Analysis

The complete sequence of Gastrimargus marmoratus mitochondrial genome was determined using long PCR and conserved primers walking approaches.Phylogenomic analysis of 30 Orthopteran species was made based on the two mitochondrial rRNA genes.1.The determined mitochondrial genome sequence of G.marmoratus is 15904 bp long,encoded 13 proteins,22 tRNAs,2 rRNAs,and an A+T rich region.The gene order is identical to Locust migratoria and other insects of Acridoidea.2.The average A+T content of the G.marmoratus mitochondrial genome protein coding sequence,rRNA gene,tRNA gene and A+T rich region were corresponding well to the A+T bias generally observed in insect mitochondrial genomes.The strongest A+T biase is found at the A+T rich region and third codon positions which are under the low purifying selection.3.A total of 45bp overlap regions among G.marmoratus mt genomes are found in ten locations with a total of 86 bp of intergenic spacer.In these ovelaps,2 of them are between different protein genes,4 between protein and tRNA genes,1 between rRNA and tRNA genes,3 between different tRNA genes.4.Common Met start codons ATN could be assigned to all protein genes,except for COI and ND5,which use TTG.Common stop codons(TAA and TAG) could be assigned to most of the putative protein sequences.Only the ND5 genes terminate into a single T residue.5.All oftRNAs showed typical clover leaf secondary structures and their anticodons are similar to those found in other Hexapoda animals,tRNAs possess invariable aminoacyl stem,anticodon loop,and anticodon stem.Most of the size variation among tRNAs stemmed from length variation in the TΨC and DHU arms.The G-U pair is found in 14 of 19 mispairs in tRNA secondary structure.6.The location of two rRNAs is conserved in insects.The predicted secondary structure of SrRNA consists of 30 stems and loops forming three domains.The first and second domains vary among species compaired,the third one is conserved.The predicted secondary structure of LrRNA contains six domains,44 hairpins(stems and loops),which varies in size among the compaired species.7.Three tandem repeats was observed in A+T rich region of Gastrimargus marmoratus mitochondrial genome.Repeat 1 and Repeat 2 with 166bp are identical.There is only one difference between the first two repeats and Repeat 3,which has 120bp.8.Phylogenetic analysis of 30 Orthopteran species were performed with maximum parsimony, maximum likelihood and Bayesian inference methods based on SrRNA data set,LrRNA data set and the combined data set.The result support the basal place of Pamphagidae,Chrotogonidae,and Pyrgomorphidae,and the monophyly of Oedipodidae,Catantopidae,and Arcypteridae.