| Insects,as the most species of animals on earth,occupy a very important position in the ecosystem.Insects often meet with various adverse abiotic stress factors during their growth and development,such as high temperature,low temperature,heavy metal,starvation and ultraviolet radiation.When they feel the adversity signal,they regulate the expression of intracellular anti-stress proteins through the signal conduction process,and adjust their own behavior or physiological state to adapt to the adverse environment.In this study,Apis cerana cerana(Chinese honeybee)was used as experimental object to study the effects on physiology and biochemistry of different times under starvation stress and Ultraviolet A(UVA)radiation.In addition,RNA-seq technology was used to further study the immune response mechanism of Apis cerana cerana under starvation stress and UVA radiation.The main conclusions of this study are as follows:1.The survival rate of Apis cerana cerana decreased significantly after different times of starvation stress;There was no significant effect of short time starvation stress(0-3 h)on the body weight of Apis cerana cerana(P>0.05)and with the extension of starvation stress(after 3 h),the body weight of Apis cerana cerana decreased significantly(P<0.05).The survival rate of Apis cerana cerana decreased significantly after different times of UVA radiation;There was no significant effect of short time UVA radiation(0-6 h)on the body weight of Apis cerana cerana(P>0.05)and with the extension of starvation stress(after 6 h),the body weight of Apis cerana cerana decreased significantly(P<0.05).2.After starvation and UVA stress for different time,the content of haemolymph protein in Apis cerana cerana increased first and then decreased,and there was a significant difference in the content of haemolymph protein between the treatment group and the control group(P<0.05).The protein content of Apis cerana cerana was the highest under starvation stress for 9 h,which was 76.58±0.60 g/L.The protein content of Apis cerana cerana was the highest under UVA radiation for 12 h,which was 93.56±2.71 g/L.3.A total clean data of 62.20 GB was obtained by transeriptome sequencing of the control group,starvation stress for 9 h and UVA radiation for 12 h.The clean data of all the 9 samples reached 6.65GB,the percentage of all the bases of Q30 was not less than 90%,and the GC content was about 40%.The ratios of clean reads to reference genomes respectively were 90.99%,91.14%,92.45%,89.54%,91.32%,92.84%,93.33%,91.14%,92.24%.4.DEseq2(|log2FC|?1 and Qvalue<0.05 as the screening criteria)was used to analyze differential expression of the obtained data:In the process of starvation stress,a total of 543 differential expression genes were screened out.The GO Function annotation results showed that respectively 298,317 and 321 genes were annotated in Biological Process,Molecular Function and Cellular Component.102 differential expression genes were enriched in response to stimulus,58 differential expression genes were enriched in signaling,and 11 differential expression genes were enriched in the immune system process.KEGG pathway analysis of differential expression genes showed that the majority of differential expression genes enriched in cellular immunity and humoral immunity were up-regulation,indicating that the immune system of Apis eerana cerana was significantly activated.5.In the process of starvation stress,a total of 585 differential expression genes were screened out.The GO Function annotation results showed that respectively 320,337 and 354 genes were annotated in Biological Process,Molecular Function and Cellular Component.117 differential expression genes were enriched in response to stimulus,61 differential expression genes were enriched in signaling,and 15 differential expression genes were enriched in the immune system process.KEGG pathway analysis of differential expression genes showed that under UVA radiation,some differential expression genes enriched in Apis cerana ceraoa cell immunity showed up-regulation trend,while lore showed down-regulation trend.The majority of differential expression genes enriched in humoral immunity were up-regulated,indicating that the cellular immune pathway of Apis cerana cerana was partially activated,while humoral immunity was significantly activated.According to the GO and KEGG results,the immune response of Apis cerana cerana under starvation and UVA stress was significantly activated to adapt to changes in the environment.6.Eight immune genes were selected from the two treatment groups and verified by qRT-PCR,and the results were consistent with RNA-seq,proving the reliability of transcriptome data. |
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