和梦颖,黎尔彤,田方圆,汪洁,金小宝,刘文彬,2023,基于转录组学的Vip3Aa杀美洲大蠊的作用机制研究[J].环境昆虫学报,45(5):1347-1355
基于转录组学的Vip3Aa杀美洲大蠊的作用机制研究
Study on the mechanism of Vip3Aa killing Periplaneta americana based on transcriptomics
  
DOI:
中文关键词:  美洲大蠊  Vip3Aa  转录组  凋亡
英文关键词:Periplaneta americana  Vip3Aa  transcriptome  apoptosis
基金项目:广东省公益研究与能力建设项目(2017A020211008);广东省自然科学基金(2020A1515011097);广州市科技计划项目(202201010357)
作者单位
和梦颖,黎尔彤,田方圆,汪洁,金小宝,刘文彬 广东药科大学生命科学与生物制药学院/广东省生物活性药物研究重点实验室广州 510006 
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中文摘要:
      美洲大蠊Periplaneta americana是常见的家居害虫,前期研究发现营养期杀虫蛋白3Aa(Vegetative insecticidal proteins 3Aa, Vip3Aa)蛋白具备杀美洲大蠊作用,但其机制尚不明确。本研究对美洲大蠊饲喂Vip3Aa蛋白后,进行中肠转录组分析,所得raw data过滤获得44.60 Gb Clean Data。通过生物信息学分析,共注释了23 470条基因,发掘新基因5 757个。与对照组比较,共获得2 302个差异表达基因,其中1 539个上调、763个下调。KEGG分析表明,差异基因主要涉及调控溶酶体、其他多糖降解、淀粉和蔗糖代谢、糖胺聚糖降解、昆虫激素合成、鞘糖脂生物合成、药物代谢和凋亡等信号通路,其中溶酶体相关差异表达基因50个,凋亡相关差异表达基因21个。透射电镜和Hoechst 33342染色显示,Vip3Aa给药后,中肠上皮细胞出现凋亡典型的形态改变,线粒体肿胀、空泡、破裂,出现大量次级溶酶体,可见部分细胞的细胞核凝集、固缩、荧光强度增加。结果表明Vip3Aa蛋白可能通过上皮细胞溶酶体-线粒体损伤介导凋亡通路的激活,发挥杀美洲大蠊作用。
英文摘要:
      Periplaneta Americana is a common household pest. Previous studies found that Vip3Aa protein can kill Periplaneta americana, but its mechanism is not clear. In this study, the midgut transcriptome of P.americana fed on Vip3Aa protein was analyzed, and 44.60 GB clean data was obtained. Through bioinformatics analysis, 23 470 genes were annotated and 5 757 new genes were discovered. 2 302 differentially expressed genes were screened, including 1 539 up-regulated genes and 763 down regulated genes. KEGG analysis showed that the differentially expressed genes were mainly related to the regulation of lysosome, other polysaccharide degradation, starch and sucrose metabolism, glycosaminoglycan degradation, insect hormone synthesis, sheath glycolipid biosynthesis, drug metabolism, apoptosis and other signal pathways. There were 50 differentially expressed genes related to lysosome and 21 differentially expressed genes related to apoptosis. Transmission electron microscopy and Hoechst 33342 staining showed that after administration of Vip3Aa, midgut epithelial cells showed typical morphological changes of apoptosis, mitochondria swelling, vacuoles and rupture, and a large number of secondary lysosomes appeared. It could be seen that the nuclear agglutination, pyknosis and fluorescence intensity of some cells increased. This study found that Vip3Aa protein may mediate the apoptosis of midgut epithelial cells of P.americana through lysosomal related signal pathway, so as to play a role in killing P.americana.
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