摘要
【目的】探究家蚕Bombyx mori肠道特异表达的金属羧肽酶基因BmMCP 18的功能及对外源病毒侵染的抵抗机制。【方法】构建敲除BmMCP18的家蚕BmMCP18KO(C18KO),对C18KO和对照组野生型家蚕(C18KOC)的5龄幼虫中肠以及感染家蚕核型多角体病毒(Bombyx mori nuclear polyhedrosis virus,BmNPV)的家蚕BmMCP18KO(C18KOV)和对照野生型(C18KOCV)的5龄幼虫中肠进行转录组测序;分析差异表达基因并进行GO功能注释和KEGG通路富集;利用qRT-PCR验证相关基因表达。【结果】与对照组C18KOC相比,C18KO有75个基因上调表达,303个基因下调表达。与对照组C18KOCV相比,C18KOV有96个基因上调表达,57个基因下调表达。C18KOC vs C18KO比较组差异表达基因显著富集的GO条目为细胞外空间、细胞表面、肽交联和突触靶标识别等。C18KOCV vs C18KOV比较组差异表达基因富集最显著的GO条目为跨膜转运蛋白活性。C18KO 5龄幼虫中肠转录组中免疫通路、碳水化合物和能量代谢通路相关基因的表达比C18KOC的显著下调,包括Toll和Imd通路及MAPK通路相关基因,而泛素介导的蛋白质水解通路相关基因表达比C18KOC的显著上调。C18KOV 5龄幼虫中肠转录组中的能量代谢基因的表达比C18KOCV的显著上调。qRT-PCR验证结果表明转录组数据可靠。【结论】BmMCP18的功能涉及家蚕中肠的细胞识别、免疫调节和能量代谢,可能是通过参与中肠细胞免疫反应、能量和物质供应,从而影响蚕体对外源病原侵染的抵抗能力。
【Aim】To explore the function of BmMCP 18,a gut-specific metallocarboxypeptidase gene from Bombyx mori,and its resistance mechanism to exogenous virus infection.【Methods】The BmMCP18 knockout B.mori(BmMCP18KO)(C18KO)was constructed.The 5th instar larval midgut transcriptomes from C18KO and its control wild-type B.mori(C18KOC),and the 5th instar larval midgut transcriptomes of BmMCP18KO(C18KOV)infected with nuclear polyhedrosis virus(B.mori nuclear polyhedrosis virus,BmNPV)and its control wild-type B.mori(C18KOCV)were sequenced.The differentially expressed genes were analyzed for GO functional annotation and KEGG pathway enrichment,and qRT-PCR was used to verify the expression of related genes.【Results】Compared with the control group C18KOC,C18KO had 75 genes with up-regulated expression and 303 genes with down-regulated expression.Compared with the control group C18KOCV,C18KOV had 96 genes with up-regulated expression and 57 genes with down-regulated expression.The significantly enriched GO items by differentially expressed genes in the C18KOC vs C18KO comparison group were extracellular space,cell surface,peptide cross-linking and synaptic target recognition.The most significantly enriched GO item by differentially expressed genes in the C18KOCV vs C18KOV comparison group was transmembrane transporter activity.The expression of genes related to the immune pathway,carbohydrate and energy metabolism pathway,including Toll and Imd pathway,and MAPK pathway in the transcriptome of the 5th instar larval midgut of C18KO,was significantly down-regulated,while that related to ubiquitin-mediated proteolysis pathway was significantly up-regulated,as compared with that of C18KOC.The expression of energy metabolism genes in the 5th instar larval midgut of C18KOV was significantly up-regulated as compared with that of C18KOCV.The qRT-PCR validation results indicated that the transcriptome data were reliable.【Conclusion】The function of BmMCP18 involves cellular recognition,immune regulation and energy metabolism in the midgut of B.mori,possibly by participating in the immune response,energy and material supply of midgut cells to affect the resistance of B.mori to the infection of exogenous pathogens.
作者
王倩茹
王亚清
沈诗桀
耿琳烨
刘东华
郭金鑫
沈以红
黄先智
WANG Qian-Ru;WANG Ya-Qing;SHEN Shi-Jie;GENG Lin-Ye;LIU Dong-Hua;GUO Jin-Xin;SHEN Yi-Hong;HUANG Xian-Zhi(State Key Laboratory of Resource Insects,Southwest University,Chongqing 400715,China;College of Fisheries,Southwest University,Chongqing 400716,China)
出处
《昆虫学报》
CAS
CSCD
北大核心
2024年第11期1572-1584,共13页
Acta Entomologica Sinica
基金
国家现代农业产业体系专项(CARS-18)。
关键词
家蚕
金属羧肽酶基因
基因敲除
家蚕核型多角体病毒
中肠转录组
Bombyx mori
metallocarboxypeptidase gene
gene knockout
Bombyx mori nuclear polyhedrosis virus
midgut transcriptome
