Transcriptome Analysis And Differentially Expressed Genes Cloning Of Apis Mellifera In Response To Ascosphaera Apis

Honey bee as an important pollinator plays a significant role in global crops and Eco-agriculture.However,diseases and environmental factors working synergistically weaken honey bee health,which may cause the losses of bee colonys.Among all of bee diseases,chalkbrood disease has been on the rise.To better explore the immune response to Ascosphaera apis,we used a clinical isolated strain of Ascosphaera apis to establish a Ascosphaera apis artificial infection model based on the bee larvae,we analyzed the transcriptome profiling of Apis mellifera challenged with Ascosphaera apis.Some immune-related genes with significantly differential expression were cloned and used for sequencing bioinformatics analysis.This study may provide precious genomic resources for future research into the immune-related genes of Apis mellifera.1.Isolation and identification of Ascosphaera apisFrom this study,clinically suspected infected bees cultivate a pathogenic fungi were isolated from sick bee larvaes,based on fungal morphology and 18 s ribosomal RNA area and molecular biology identification of universal primers,we determined disease pathogenic fungi was Ascosphaera apis,the separation of pathogenic bacteria identification will provide the basis for subsequent experimental study.2.Transcriptome Analysis of Apis mellifera In Response To Ascosphaera apisWe obtained 50,175,666 and 42,001,818 unigenes through Illumina RNA sequencing and sequence assembly,2890 differentially expressed genes were found from DGEs ibraries.2214 up-regulated genes and 676 down-regulated genes in the comparison of healthy honey bee larvae and chalkbrood disease larvae were found by DEGs significant analysis.We did Gene Ontology(GO)enrichment analysis and Pathway enrichment analysis.Immune-related genes diffierent expression,main gene families and related pathways in Ascosphaera apis pathogen pressure of honey bees were clearly screened at the transcriptome level.Using the transcriptome analysis,several key transcriptional pathways were found that compose the whole effort in the honey bee larvae to resist Ascosphaera apis infection.In this study,immune regulation of the Ascosphaera apis responses in honey bee larvae was found to involve highly synergistic activation in Jak-STAT signaling pathway,NF-?B signaling pathway and Toll-like receptor signaling pathway.These pathways leading to the reaction of anti-microbial and the production of anti-microbial peptides.3.Cloning and analysis of differentially expressed genes in Apis melliferaAccording to the transcriptome sequencing,significantly differentially expressed genes data have been obtained,designing specific primers to bees myeloid differentiation factor(myeloid differentiation factor 88,MyD88),Abaecin antibacterial peptide gene family:Abaecin,Hymenoptaecin,Defensin 1 gene sequence length on CDs amplification and build a cloning vector,phylogenetic analysis was carried out on the related sequence,amino acid composition,index,the antigen epitope antigen,secondary structure and tertiary structure prediction of bioinformatics analysis,related genes and proteins function for the further research in the future to provide a oretical reference.