| The codling moth,Cydia pomonella(L.)is a kind of destructive quarantine pest which is widely distributed in the world.It is harmful to the fruit borer of larva and causes great damage to the global fruit industry.Because of the long-term dependence on chemical insecticides for control,the problem of resistance to insecticides is becoming more and more serious in many countries and regions,and the cost of prevention and control and the hidden danger of quality and safety are increasing.It is an effective way to open up a new control method for apple moth to study the mechanism of resistance to insecticides and dig out a new target.It has become a research hotspot rapidly and the research difficulty has been greatly reduced by the completion of genome sequencing of the apple moth.The success rate is improved.In this paper,based on the genomic data of the apple moth,the genes related to insecticide resistance in the genome of the moth were deeply excavated by bioinformatics and molecular biological techniques.Based on the data of re-sequencing,11 resistance-related gene families were obtained,and GWAS analysis of the key genes was carried out.In order to fully analyze the resistance mechanism of the moth,a new target was found.Scientific data and theoretical basis were provided for the development of new insecticide for effective integrated pest management program.1.Based on genomic data of the codling moth,11 resistance-related gene families were obtained from high throughput analysis of insecticide resistance genes at the genomic level.By genome search and gene family analysis,127 cytochrome P450 genes,21 glutathione mercapto transferase genes,74 carboxylesterase genes,66 ABC transporters,4 glutamic gated chloride channels,4 GABA gated chloride channels,1 calcium ions,1 voltage-gated sodium channels,12 nicotinic acetylcholine receptors,2 acetylcholinesterase genes and 11 aquaporin coding genes were obtained in the genome of the codling moth,together with a total of 11 gene family related to pesticide resistance.Comparative analysis with other species revealed that the P450 gene,ABC transporter,and aquaporin were significantly amplified.2.Target resistance is an important mechanism of insect resistance.The nucleic acid and protein sequences of 12 nicotinic acetylcholine receptor genes were obtained from the genome data of codling moth,in order to further study the codling moth to neonicotinoid insecticides.The agent lays the foundation for the target resistance mechanism.The full-length cDNA sequences of the 8 nAChR subunit genes of codling moth were cloned using modern molecular biology techniques,and the bioinformatics analysis of the sequence structures of the eight-subunit genes was performed.The molecular weights of the eight nAChR subunit genes of the codling moth are between 36-60 kD,and the PI value is between 4.7 and 8.0.Both contain ?-helices,?-sheets,extended chains,and random coils,and contain introns.The number of exons is not the same,and all gene sequences are conserved.3.The detoxification function of cytochrome P450 is also an important mechanism of insect resistance.Through 5?RACE cloning of 127 P450 genes in the genome of the codling moth,the 5?UTR regions of 100 P450 genes were successfully obtained and mapped to the genome,and GWAS was performed using the re-sequencing data of codling moth.A total of five SNP sites related to methyl-glutathione resistance were detected in the promoter regions of the 100 P450 gene.Forty nine SNP sites related to deltamethrin resistance were detected.Most of these P450 genes were members of the Clan3 and Clan4 subfamily.In the coding regions of other key genes such as CarE,ABC,and GST,we found four SNP sites associated with methyl glutamate resistance and twenty four SNP sites associated with the resistance of deltamethrin. |
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