基于Illumina HiSeq测序技术研究吡虫啉对意大利蜜蜂雄蜂的影响  

Using Illumina HiSeq sequencing technology to reveal the effects of imidacloprid on Apis mellifera Ligustica drones

作　　者：李剑飞 庄明亮[1] 李志勇[1] 牛庆生[1] 王志[1] 葛蓬[1] 张发[1] LI Jian-Fei;ZHUANG Ming-Liang;LI Zhi-Yong;NIU Qing-Sheng;WANG Zhi;GE Peng;ZHANG Fa(Apiculture Science Institute of Jilin Province,Jilin 132108,China)

机构地区：[1]吉林省养蜂科学研究所,吉林132108

出　　处：《应用昆虫学报》2021年第5期1103-1112,共10页Chinese Journal of Applied Entomology

基　　金：吉林省科技发展计划项目-自然科学基金(20190201284JC)。

摘　　要：【目的】雄蜂对蜂群繁衍有着非常重要的作用。本研究旨在探究吡虫啉对意大利蜜蜂Apis mellifera Ligustica雄蜂生长发育和基因表达产生的影响。【方法】以意大利蜜蜂雄蜂为研究对象,分别以0.00001、0.0001和0.001μg/μL浓度的吡虫啉对雄蜂幼虫进行连续饲喂处理。每天观察并记录幼虫的发育形态及死亡率,在雄蜂幼虫后期(移虫后6d)测量幼虫体重。利用Illumina HiSeq测序技术对经吡虫啉处理的雄蜂进行转录组测序,进而对差异表达基因进行深入分析。【结果】取食吡虫啉后的雄蜂幼虫,体重低于正常雄蜂,当浓度高于0.0001μg/μL时差异显著;雄蜂幼虫取食吡虫啉后出现死亡现象,且死亡率随吡虫啉浓度的升高而增大;差异表达基因分析结果上调与下调基因数量分别为390个和130个。GO富集分析结果上调基因共分布于55个GO条目,富集基因数量最多的是细胞进程、细胞、细胞组件、细胞膜、细胞膜组件、结合,下调基因共分布于48个GO条目,富集基因数量最多的是细胞进程、细胞、细胞组件。富集在有关生殖功能的差异表达基因中,上调基因数量为21个,下调基因数量为5个。KEGG代谢通路富集分析结果上调基因富集在159个通路上,其中富集基因数最多的是蛋白质消化吸收和神经活性配体-受体相互作用通路。下调基因富集在71个通路上,其中富集基因数最多的是溶酶体、胰液分泌、神经活性配体-受体相互作用通路。【结论】吡虫啉能抑制意大利蜜蜂雄蜂的生长发育,甚至造成幼虫死亡,同时,可以影响雄蜂的神经系统、代谢系统和生殖系统等。本研究结果为蜜蜂资源保护提供理论依据。[Objectives]To investigate the effect of imidacloprid on the growth,development and gene expression of Apis mellifera Ligustica drones.[Methods]A.mellifera drone larvae were fed solutions containing 0.00001,0.0001 or 0.001μg/μL imidacloprid and their development and mortality were recorded daily.Larval weight was measured in the late larval stage(6 d)and differential expression of genes was analyzed in depth.[Results]Imidacloprid treated drone larvae were lighter in weight than control larvae,significantly so at concentrations higher than 0.0001μg/μL.Mortality was also higher in the imidacloprid treatment group and increased with dosage.Analysis of differential gene expression indicated that 390 genes were up-regulated and 130 down-regulated.GO enrichment analysis revealed that the up-regulated genes were distributed in 55 GO teams,most of which are involved in cellular process,cells,cell parts,membranes,membrane parts and binding.Down-regulated genes were distributed in 48 GO teams,most of which are involved in cellular processes,cells and cell parts.There were also differentially expressed genes involved in reproduction,21 of which were upregulated and 5 of which were down-regulated.KEGG pathway enrichment analysis showed that up-regulated genes were enriched in 159 pathways,among which the most enriched were involved in protein digestion and absorption and neuroactive ligand-receptor interaction.Down-regulated genes were enriched in 71 pathways,among which the most enriched were involved in lysosome,pancreatic secretion and neuroactive ligand-receptor interaction.[Conclusion]Imidacloprid can inhibit the growth and development of A.mellifera drones,and can cause larval mortality.Analysis of gene expression indicates that imidacloprid affects the nervous,metabolic and reproductive,systems of drone larvae.These results highlight the importance of protecting bees from the toxic effects of pesticides.

关 键 词：意大利蜜蜂 雄蜂 吡虫啉 转录组 

分 类 号：S891[农业科学—特种经济动物饲养]