| Apis mellifera ligustica is the main bee species raised in China and plays an indispensable role in maintaining the balance of the ecosystem and the sustainable development of the ecological environment.Compared with others,Apis mellifera ligustica has weaker anti-mite ability.In recent years,bee mites have become a major enemy restricting the development of Chinese bee industry due to their wide range and high degree of damage,causing huge losses to the Chinese bee industry.At present,the main component of commonly used acaricides is fluvalinate,which is a widely used and relatively safe acaricide for bees,but it will still have a negative impact on the colony after use;As a kind of non-coding small RNA,microRNA can participate in various biological regulation processes at the post-transcriptional level,and participate in many biological functions in the organism.In the experiment,6 groups of Apis mellifera ligustica with similar group potential were randomly selected,among which 3 groups were blank control groups(three replicate groups),and 3 groups were experimental groups(each group was three replicate groups).The experimental group selected fluvalinate strips for administration,dissected the brain tissue and performed miRNA sequencing,and analyzed the differentially expressed microRNAs and potential target genes in the brain tissues of fluvalinate over time after the use of fluvalinate,in order to reveal the role of differentially expressed miRNA in the response to fluvalinate stress.The research results are as follows:1.Twelve samples were sequenced to obtain a total of 184051902 clean reads.After filtering the data,the final s RNA clean tags sequence obtained was 18109705 at most and9626768 at least,with a distribution range of 94% to 98%.The number of reads in each sample library is within a reasonable range,there are no abnormal samples,and the sequencing quality is good.2.After annotation,a total of 1069 miRNA were identified to be expressed in 12 Apis mellifera ligustica brain tissues;the differentially expressed miRNAs were screened from the samples treated with fluvalinate stress and the control group(FDR<0.05 and|log2(FC)| >1)15,including two novel miRNAs(novel-m0194-5p,novel-m0184-5p).Among these differentially expressed miRNAs,CK-vs-X1D10 had the most up-regulated differential expression,with a total of 6;CK-vs-X1D20 had the most down-regulated differential expression,with 7 of them.3.The cluster analysis of expression patterns showed that different miRNAs showed three expression patterns(C1,C2,C3).Among them,clusters C1 and C2,which accounted for more than half of the total number of differential miRNAs,generally showed high expression after medication,with a peak on the 10 th day,and with the increase of medication time,the expression level gradually decreased to the level before medication;and mainly concentrated in various types Metabolism-related pathways indicate that the transcription level regulation involved in C1 and C2 cluster miRNAs may be related to stress response;after C3 clusters account for nearly half of the total number of differential miRNAs,the expression of miRNAs gradually declines and becomes normal after 20 days.It is mainly enriched in functional pathways,indicating that the transcriptional level regulation involved in these differential miRNAs may be related to the response process.4.In the GO enrichment analysis,biological processes are mainly concentrated in the process of cell protein modification,biological regulation,and m RNA metabolism;cell components are mainly distributed in cells and organelles;molecular functions are mainly distributed in binding and cell protein modification processes.Through enrichment analysis,we found that the target genes corresponding to differential miRNAs were significantly enriched in three KEGG pathways,namely Dorso-ventral axis formation pathway,Phototransduction-fly pathway and Wnt signaling pathway.The results of this study proved that the miRNA expression profile in the brain did change correspondingly after the use of fluvalinate by bees.The functional enrichment results indicated that these differential miRNAs may be involved in the apoptosis process of the hippocampal cells and brain memory.The regulation of the process of functional impairment may be related to the behaviors of the Apis mellifera ligustica not returning home at night and behavior disorder after medication.In general,this study provides new information for the study of the molecular mechanism and related pathways of fluvalinate on the brain tissue of Apis mellifera ligustica. |
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