Idetification And Differetial Analysis In Cytochrome P450 Of Pardosa Astrigera Based On Transcriptome Sequencing

Pardosa astrigera is the important predators of insect and play the crucial role in biological control.A series of insecticides was used to kill insects and cause the "3R"phenomenon,while the spider is affected.So that the aim of the present study was to understand the mechanism of pesticides on Pardosa astrigera,Therefore,The transcriptome sequencing technology was employed to sequence the Pardosa astrigera transcriptome in order to expand the gene information resources of the species.Screening Pardosa astrigera cytochrome P450 and differentially expressed genes,provide a theoretical basis for the development of resistance to the dominant enzyme inhibitors.and lay a molecular foundation for future study of insecticide resistance.The main results as follows:1.The experiment using deltamethrins pharmacodynamics results handle the Pardosa astrigera because of the longer life span of Pardosa astrigera about 165 days.The results showed that the concentration of LC10 was 5.151mg·L-1,LC30 was 8.619 mg·L-1,LC50 was 12.311 mg·L-1.The toxicity of deltamethrin is weaker than cyhalothrin(LC50 was 3.2 mg·L-1)and avermectin(LC50 was 7.51 mg·L-1).So,sexually mature Pardosa astrigera treated with deltamethrin three concentrations were subjected to transcriptome sequencing.2.Our project is sequenced on the platform of Illumina Hiseq 2000.19,468,934,460nt bases are generated totally.In the results of assembly,68410 Unigenes are detected,total length for Unigenes is 74,731,570 nt,average length is 1092 nt,N50 is 2048 nt.Q20 was higher than 98%,the ratio of the number of bases that could not be determined to the total number of bases was 0.01%,the GC content was 39.02%,and the average N50 of conting and Unigene was 419 nt and 1500 nt.For function annotation analysis,we get 24713,10365,21300,18701,10225,12255 Unigenes which annotate to the NR,NT,Swiss-Prot,KEGG,COG,GO database,respectively,the total annotation Unigenes are 26814.For protein coding region prediction analysis,the number of CDS that mapped to the protein database is 24688,the number of predicted CDS is 3367.The total number of CDS is 28055.SSR is 10869.3.On the basis of transcriptome,manually searching the database.Use ORF finder to translate Unigene,The conserved amino acid sequence was used to identify the cytochrome P450,and the phylogenetic tree was used to classify the identified cytochrome P450 by using the Neighbor-joining(NJ)method in MEGA5.Result shows that,a total of 66 P450 genes were identified,and 23 were associated with CYP2 family gene,11 were associated with CYP3 family gene.There were 17 belong to CYP4 and 15 others belonged to other families(CYP10,CYP12,CYP49,etc).The P450 genes were compared to the KEGG database and found a total of 13 metabolic pathways,and some of them were associated with detoxification.4.Differentially expressed genes were screened with different concentration treatments Pardosa astrigera,adopting FPKM calculation method.The criteria of false discovery rate(FDR)?0.001 and absolute value of log2 ratio?1 were used to identify differentially expressed genes(DEGs).We selected 10 cytochrome P450 Unigenes which were differentially expressed genes.GO and KEGG pathway enrichment analysis revealed that the 5 P450 DEGs were significantly enriched in GO and KEGQ and the GO term and KEGG pathways were involved in some metabolic activities,all related to the metabolism of exogenous substances,so the final screening of five star leopard spider cytochrome P450 differentially expressed.Then predicted they were associated with resistance.