阿尔泰蝠蛾(鳞翅目:蝙蝠蛾科)幼虫低温胁迫转录组分析

Gene transcription in Hepialus altaicola(Lepidoptera:Hepialidae)larvae subject to cold stress

【目的】本研究旨在通过转录组测序技术分析低温胁迫引起的阿尔泰蝠蛾Hepialus altaicola Wang幼虫基因转录差异及上调表达基因的主要功能类群和参与的主要代谢通路。【方法】采用Illumina HiSeqTM 2500测序平台对阿尔泰蝠蛾幼虫进行转录组测序、组装,利用Blast软件进行数据库比对和基因功能注释,用DEGSeqR软件包分析4℃低温处理与室内适温饲养试虫的差异表达基因,并对上调表达基因进行GO和KEGG代谢途径富集分析。【结果】经序列拼接后共获得100300个unigenes,总长度81600309 bp,平均长度813 bp,N50长度1719 bp。与7大数据库同源比对,共获得34691(34.59%)条unigenes。低温胁迫转录组分析得到11569个差异表达基因(DEGs),7158条基因上调,4411条基因下调。富集到47个GO类群,217个KEGG途径。其中代谢过程、催化、结合活性类群占有重要比例,核糖体、碳代谢、剪接体等途径显著富集。另外,热激蛋白、昆虫表皮蛋白、海藻糖酶、超氧化物歧化酶等非生物胁迫相关基因显著上调表达。【结论】阿尔泰蝠蛾幼虫低温胁迫转录组分析揭示,代谢过程、细胞过程、生物调节、对刺激的反应等生物学过程相关基因和部分非生物胁迫响应基因显著上调表达,提示蝠蛾幼虫可能从抗氧化防御、分子伴侣、体温调节和维持细胞的渗透平衡等多方面应对低温胁迫。

[Objectives]To use transcriptome sequencing to identify the main functional groups of up-regulated genes,and the main metabolic pathways of Hepialus altaicola larvae,and investigate the effects of cold stress differences on gene transcription.[Methods]The transcriptome of H.altaicola larvae was sequenced and assembled on the Illumina HiSeq;2500 sequencing platform.Database comparison and gene annotation were conducted using Blast software.The DEGSeq R software package was used to analyze differentially expressed genes of larvae kept at 4℃ and GO term enrichment and KEGG metabolic pathway analysis were performed on the up-regulated genes.[Results]After sequence splicing,a total of 100300 genes were obtained with a total length of 81600309 bp,a mean length of 813 bp and an N50 length of 1719 bp.Comparison with seven major databases identified 34691(34.59%)unigenes.Transcriptome analysis revealed 11569 genes that were differentially expressed in response to cold stress(DEGs),7158 of which were up-regulated and 4411 down-regulated.The up-regulated genes were enriched to 47 GO terms and 217 KEGG pathways.An important proportion of these were involved in the metabolic process,catalysis and binding activity teams.Ribosome,carbon metabolism,spliceosome and other pathways were also significantly enriched.In addition,abiotic stress-related genes,such as heat shock protein,insect cuticle protein,trehalase and superoxide dismutase,were up-regulated.[Conclusion]The transcriptomic data of H.altaicola larvae reveal that the expression of genes related to metabolic processes,cellular processes,biological regulation,response to stimuli,as well as some abiotic stress response genes,are significantly up-regulated in response to cold stress.This suggests that H.altaicola larvae use a range of strategies to cope with cold stress,including antioxidant defense,molecular chaperones,body temperature regulation and maintenance of cell osmotic balance.