Comparative Transcriptomic And Proteomic Analysis Of Pupation Of Apis Mellifera Ligustica Under Low Temperature Stress

The honey bee is among the holometabolous insects which brood is typically stenothermic.In this study,In order to explor low temperature on pupation's damage and key gene that involved in honey bee Apis mellifera ligustica 3-day-old worker brood sensitivity to cold stress.we conducted the statistics of pupation rate,the observation of the changes of external morphology and fat body during pupation under low temperature stress 18 h and 36 h.Meantime,the differential expression genes and differential proteins of 3-day-old worker brood under low temperature stress at different times were analyzed by means of transcriptomics and proteomics.Further,q RT-PCR was used to detect gene response reactions on key pathways affected by low temperature stress.This research shows are as follows:(1)Apis mellifera ligustica pupation is affected by low temperature.The external morphology showed that the pupation failed under low temperature stress,and when restored to 35?,The pupation rates of T18 and T36 were 81.71% and 9.23%,the pupation rate decreased with the extension of stress time.TUNEL staining showed that fat body did not recombine and differentiate at low temperature,only apoptosis occurred,the apoptosis rates were 24.16% and 40.20% respectively,which increased with the prolongation of low temperature stress time.(2)The transcriptome data showed that most of the differentially expressed genes were enriched in metabolic pathways.At the same time,pupation related genes was detected,such as CREB-binding protein(CBP),CYP450 306a1(CYP306a1),insulin-like peptide(ILP),forkhead box protein O(Fox O)et al..phm has the function of regulating 20 E synthesis,The decrease of its expression level may lead to the decrease of 20 E synthesis titer.Moreover,Under cold stress,Fox O phosphorylated by ILP and 20 E in honey bee were arrested in cytoplasm repressing the transcription activity of Fox O in the nucleus that fails to prime downstream genes in 20 E signaling and ecdysis.(3)q RT-PCR was used to detect the gene response in Hippo signal pathway,Wnt signal pathway,MAPK signal pathway,Fo XO signal pathway.The results showed that the expression of most genes in the pathway was inhibited under low temperature stress,which may affect tissue and cell development during pupation,such as recombination,differentiation,autophagy and apoptosis.(4)Proteome data showed that differentially expressed proteins' number increased significantly with the extension of low temperature stress time,and most were concentrated on signal pathways such as protein synthesis and transportation,cell development and hormone synthesis.(5)Association analysis of proteome and transcriptome shows that both cytochrome P450 4C1(CYP4C1)and cytochrome P450 4C1(CYP4C1)are down-regulated expression which has the function of regulating juvenile hormone synthesis.Therefore,low temperature may lead to an increase in JH level and inhibit 20 E synthesis,thus leading to the inability of 3-day-old worker brood to ecdysone.The results of this study can enrich the mechanism of temperature influence on the development of worker brood and provide basic data for further study of its mechanism.It is helpful to study the feedback and damage of insects and even life to low temperature.It is helpful to understand the different survival strategies of social insects and solitary insects in response to environmental temperature changes.It is of great significance to enrich insect ecology and insect developmental biology.