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目的 通过甲基化测序结合生物信息学分析，筛选感染与未感染CSBV中蜂幼虫的差异表达基因，鉴定出中华蜜蜂20-蜕皮激素单氧酶基因（20-Ecdysone Monooxygenase, 20emo），探究其在中蜂感染中华蜜蜂囊状病毒（Chinese sacbrood virus, CSBV）过程中的功能，为CSBV防控提供潜在分子靶点。方法 采用实时荧光定量PCR(qPCR)验证20emo的表达变化，进行时空表达分析；利用RNAi干扰技术敲低20emo，检测CSBV复制及相关基因表达；对20emo基因序列及编码蛋白进行分析。结果 筛选出8个激素合成分泌相关差异基因，20emo在CSBV感染早期的表达被显著抑制；20emo在中蜂不同发育阶段表达差异显著，成蜂肠组织表达量最高；敲低20emo后CSBV复制受到抑制，IMD、TOLL免疫通路相关基因表达上调，营养应激相关基因表达也发生改变；20emo编码蛋白属细胞色素P450家族，与西方蜜蜂同源蛋白功能相近。结论 20emo可能通过调控中蜂激素平衡与免疫应答影响CSBV复制效率，为揭示蜜蜂生长发育基因的抗病毒作用提供新视角，也为CSBV防控提供了潜在分子靶点。
Abstract：Objective Using methylation sequencing combined with bioinformatic analyses, we screened differentially expressed genes in Apis cerana larvae infected or uninfected with Chinese sacbrood virus(CSBV). We identified the A. cerana 20-ecdysone monooxygenase gene(20 emo) and investigated its functional role during CSBV infection, aiming to provide potential molecular targets for CSBV prevention and control.Methods Quantitative real-time PCR(qPCR) was used to validate changes in 20 emo expression and to analyze its spatiotemporal expression patterns. RNA interference(RNAi) was employed to knock down 20 emo, followed by assessment of CSBV replication and the expression of immune-and metabolism-related genes. In addition, sequence and functional analyses of the 20 emo gene and its encoded protein were performed.Results Eight differentially expressed genes associated with hormone synthesis and secretion were identified, and 20 emo expression was significantly downregulated at the early stage of CSBV infection. The expression of 20 emo varied markedly across developmental stages of A. cerana, with the highest levels observed in the intestinal tissue of adult bees. Knockdown of 20 emo resulted in suppressed CSBV replication, accompanied by upregulation of genes involved in the IMD and Toll immune pathways, as well as altered expression of genes related to nutritional stress. The protein encoded by 20 emo belongs to the cytochrome P450 family and exhibits functional similarity to its homolog in Apis mellifera. Conclusion The 20 emo gene may influence CSBV replication efficiency by regulating hormonal homeostasis and host immune responses in Apis cerana. These findings provide new insights into the antiviral roles of honeybee growth-and development-related genes and identify 20 emo as a potential molecular target for CSBV prevention and control.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.250343
中图分类号:S895
引用信息:
[1]戚杨,黄滢铄,刘宇明,等.中华蜜蜂20-蜕皮激素单氧酶通过调控宿主免疫影响CSBV复制的研究[J].病毒学报,2026,42(01):269-280.DOI:10.13242/j.cnki.bingduxuebao.250343.
基金信息:
国家自然科学基金（项目号：32172789）,题目：m6A修饰在中蜂囊状幼虫病毒复制及RNAi免疫调控中的作用与机制研究; 辽宁省教育厅高校青年基金项目（项目号：LJ212410160014）,题目：西方蜜蜂幼虫特异性肠道共生菌抗中华蜜蜂囊状幼虫病毒机制的研究; 科技创新团队项目（项目号：LJ222410160007）,题目：重要动物疫病和人兽共患病信息库及其防控体系建立~~