RNAi Analysis Of Albendazole Resistance-related Genes In Haemonchus Contortus

In order to clarify the molecular mechanism of albendazole resistance in Haemonchus contortus and verify the function of resistance related genes,this study was based on the comparative analysis of transcriptome sequencing data of albendazole-sensitive and albendazole-resistant strains of Haemonchus contortus,resistance-related genes in the four signaling pathways necessary for the Dauer growth and development of Haemonchus contortus were screened,and bioinformatics analysis was carried out on the screened differentially expressed genes.The gene silencing of x L3 significantly different genes and drug resistance-related genes GST,P450 and ABC in Haemonchus contortus was carried out by RNAi,and the duration of drug resistance-related gene silencing with significantly reduced expression after interference was tested.For the drug resistance-related genes with significantly reduced x L3 expression after RNAi,carrying out the RNAi experiment of worm eggs of Haemonchus contortus,and the interference temperature of worm eggs was optimized.The drug resistance correlation of worm eggs after interference was verified by the incubation experiment of worm eggs.The results are as follows:（1）In the Dauer signaling pathway of Haemonchus contortus,there were 101differential genes in the c GMP signaling pathway,and GCY-12 was selected as the candidate gene for drug resistance for subsequent studies.There are two differential genes in the ILS signaling pathway,and there is no significant differential gene in this pathway.There were 32 differential genes in the TGF-βsignaling pathway,and RSKS-1 was selected as the candidate gene for drug resistance for subsequent studies.There were 22differential genes in the hormone-like signaling pathway,and UGT2B31 was selected as the drug resistance candidate gene for subsequent studies.（2）The expression of P450,RSKS-1 and UGT2B31 did not change significantly after x L3 RNAi.The expression of ABC increased significantly,the expression of GST and GCY-12 decreased by 46%and 61.5%respectively.The silencing time of GST and GCY-12 was 9d and 11d respectively,which can be used for subsequent drug resistance studies.（3）The optimal temperature for RNAi of Haemonchus contortus eggs was 12°C.The EC₅₀of the egg hatching test treatment group after GST interference was 70.22%lower than that of the blank control group.The EC₅₀of the egg hatching test treatment group after GCY-12 interference was 70.86%lower than that of the blank control group,the sensitivity was increased.