Identification And Prokaryotic Expression Analysis Of UDP-glycosyltransferases In Lissorhoptrus Oryzophilus Kuschel

Lissorhoptrus oryzophilus Kuschel belongs to the genus Coleoptera,which are important quarantine pests.Chemical pesticides are mainly used for prevention and control in paddy fields,which not only easily cause resistance of rice water weevil,but also cause pesticide residues and environmental pollution.The olfactory recognition system of insects is closely related to their behaviors of feeding and avoiding harm.Therefore,starting from the study of olfactory related genes,the research on the gene of odor-degrading enzyme of rice water weevil can provide new ideas for the prevention and treatment of rice water weevil.UGTs are widely distributed and can catalyze glycosylation reaction.They are very important metabolic enzymes in insects.They mainly catalyzes the transfer of glycosyl groups from the activated udp-glucose donor to the receptor hydrophobic small molecule-glycoside ligand,so that the hydrophobic molecules can form more hydrophilic compounds for efficient degradation and discharge.According to research,UGTs can degrade small odor molecules and play an important role in olfactory system.It is one of the potential odor-degrading enzymes in insects.In this study,13 rice water weevil UGTs(LoryUGTs)genes were screened and identified based on the antennae transcriptome data of rice water weevil.13 genes were compared with known UGT odor-degrading enzyme genes.Through bioinformatics analysis methods combined with constructing phylogenetic evolution tree,the results showed that the LoryUGTs with glycosyl transferase major structure domain and related function points are closely related to coleoptera,Holotrichia parallela UGTs and Tribolium castaneum UGTs.Therefore,LoryUGTs have the main function of glycosyltransferase.Real-time fluorescence quantitative PCR was used to detect the expression patterns of LoryUGTs gene in head,chest,abdomen,foot and antenna tissues.After the qRT-PCR detection,LoryUGTs genes are highly expressed in antennae,the relative expression levels of LoryUGT90778 and LoryUGT76491 were significantly different between antennae and other tissues,and were specifically expressed in antennae.Through bioinformatics analysis,LoryUGTs were compared with UGT2B7,a known uridine diphosphate glucose transferase gene,on the basis of similar primary structure,the three-dimensional crystal structure of UGT2B7 protein in PDB protein database was used as the template,LoryUGTs were homologous modeling and their binding sites were analyzed,after the models are evaluated and qualified,their three-dimensional structures are analyzed.The results showed that the three dimensional structure of LoryUGT90778 and LoryUGT76491 were similar to UGT2B7.It is speculated that LoryUGT90778 and LoryUGT76491 also have some functional similarities with UGT2B7.Since UGT2B7 is a known udp-glycosyltransferase.It plays a role in odor degradation,therefore,it was speculated that rice water weevil udp-glycosyltransferase gene was a potential odor-degrading enzyme,which played an important role in the olfactory recognition mechanism of insects.To further clarify the function of rice water weevil UGTs genes,two genes LoryUGT90778 and LoryUGT76491 were cloned,escherichia coli vector was constructed and SDS-PAGE was used to detect the expression of the target protein.It was used for functional analysis,laying a foundation for further analysis of the function of LoryUGTs gene.