野蛞蝓对高低温胁迫响应的转录组分析  

Transcriptome analysis on response of Agriolimax agrestis to high and low temperature stress

作　　者：王艺焜 周建云 饶陈 冯裕洋 祝紫江 祖庆学 WANG Yi-kun;ZHOU Jian-yun;RAO Chen;FENG Yu-yang;ZHU Zi-jiang;ZU Qing-xue(Guiyang Branch Company,Guizhou Tobacco Company,Guiyang 550025,China)

机构地区：[1]贵州省烟草公司贵阳市公司,贵阳550025

出　　处：《西南农业学报》2025年第1期115-124,共10页Southwest China Journal of Agricultural Sciences

基　　金：贵州省烟草公司贵阳市公司科技项目(2022-07)。

摘　　要：【目的】探究野蛞蝓对极端温度的适应机制,为野蛞蝓防治提供参考依据。【方法】以常温处理(25℃)为对照(TC),采用Illumina高通量测序技术,分别对经过35℃高温(TH)胁迫1 h、-2℃低温(TL)胁迫1 h及TC的野蛞蝓成体进行转录组测序分析;筛选不同温度处理野蛞蝓的差异表达基因,并对其进行KEGG相关代谢通路富集分析与功能注释;采用实时荧光定量PCR(qRT-PCR)对筛选的热激蛋白基因进行验证。【结果】对比组合TH vs TC的差异表达基因为1127个,包括531个表达上调,596个表达下调;对比组合TL vs TC的差异表达基因为1003个,包括631个表达上调,372个表达下调;对比组合TH vs TL的差异表达基因为1847个,包括811个表达上调,1036个表达下调。GO富集表明,差异表达基因主要与氧化还原过程、蛋白水解、细胞组分组织或生物发生和细胞蛋白代谢过程相关。KEGG富集表明,差异基因主要参与内质网蛋白质加工、核糖体合成、长寿调节途径-多物种和抗原处理及呈现等途径。与温度胁迫相关的2类基因热激蛋白和细胞色素P450均表现为上调,表明其可能参与野蛞蝓对极端温度的耐受性作用。随机筛选的HSP70、HSP16.6、HSP90等7个热激蛋白基因qRT-PCR测定结果与转录组测定结果一致,表明转录组测序数据准确可信。【结论】本研究筛选得到野蛞蝓响应温度胁迫的关键基因HSPs和P450,其差异表达可能在抵御高低温胁迫中发挥重要作用。研究结果有助于进一步理解野蛞蝓响应高低温胁迫的分子机制,为野蛞蝓的防治提供参考依据。【Objective】The adaptation mechanism of Agriolimax agrestis to extreme temperature was studied to provide a theoretical basis for control of A.agrestis.【Method】The transcriptomes of A.agrestis adults at high temperature(TH,at 35 ℃ for 1 hours),low temperature(TL,at-2 ℃ for 1 hours) and normal temperature(TC,25 ℃) were detected by Illumina high-throughput sequencing.The functional annotation and KEGG relative metabolic pathway of the differential expression genes were analyzed.The screened heat shock protein was verified by real-time fluorescence quantification PCR(qRT-PCR).【Result】There were 1127 differential expression genes(531 up-regulated genes and 596 down-regulated genes) in TH vs TC combination.There were 1003 differential expression genes(631 up-regulated genes and 372 down-regulated genes) in TL vs TC combination.There were 1847 differential expression genes(811 up-regulated genes and 1036 down-regulated genes) in TH vs TL combination.GO enrichment analysis showed that the differential expression genes were mainly related to oxidation-reduction process, proteolysis, cell component tissue or biogenesis and cellular protein metabolic process.KEGG enrichment analysis revealed that the differential expression genes involved in pathways of antigen treatment and presentation, endoplasmic reticulum protein processing, longevity regulation pathway-multispecies and ribosome.Heat shock protein and cytochrome P450 genes both were up-regulation, which indicated that these genes may involve in the tolerance of A.agrestis to extreme temperature.The qRT-PCR results of seven randomly screened heat shock protein genes(HSP70,HSP16.6,HSP90 and so on) were consistent with the transcriptome results, which indicated that the transcriptome sequencing data was accurate and credible.【Conclusion】The screened key genes HSPs and P450 are related to response of A.agrestis to temperature stress and their differential expression may play an important role in resisting high and low temperature stress of A.agrestis.

关 键 词：野蛞蝓 转录组分析 温度胁迫 热激蛋白 GO富集 KEGG富集 

分 类 号：S433[农业科学—农业昆虫与害虫防治]