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为了研究中蜂囊状幼虫病毒（Chinese sacbrood virus,CSBV）感染中华蜜蜂幼虫后体内MicroRNAs(miRNA)表达谱变化，探讨差异miRNAs(DEmiRNAs)在中蜂幼虫响应CSBV应答中的功能。本研究分别对感染CSBV24h和48h中蜂幼虫进行Illumina高通量测序，通过生物信息学软件分析DEmiRNAs及其靶基因，并进行GO功能注释和KEGG通路富集分析。通过RT-qPCR对测序结果进行验证。结果显示：对CSBV感染前后中蜂幼虫测序共筛选出140个已知miRNAs和预测到5个新miRNAs。其中，L24I组（CSBV感染24h幼虫）和L24H组（健康幼虫）对比发现18个DEmiRNAs，其中有12个表达上调，有6个表达下调；L48I组（CSBV感染48h幼虫）和L48H组（健康幼虫）筛选到47个DEmiRNAs，其中有30个表达上调，有17个表达下调。GO分析表明，部分DEmiRNAs及相关靶基因参与免疫系统过程和刺激反应等功能；KEGG通路分析显示，除涉及物质和能量代谢通路外，主要的免疫通路包括MAPK、溶酶体、Hippo、凋亡信号通路等。选取对蜜蜂调控具有重要作用和将要研究的8个miRNAs进行RT-qPCR验证结果表明，结果表明其表达水平与测序结果高度一致，表明测序准确可靠。通过本研究，不仅有助于我们深入认识CSBV感染中蜂幼虫过程中宿主miRNA变化情况，也为探究中华蜜蜂和CSBV相互作用时miRNA发挥的功能提供帮助。
Abstract：In order to study the changes of MicroRNAs(miRNA) expression profile in Apis cerana larvae infected with CSBV, the function of differential miRNAs(DEmiRNAs) in Apis cerana larvae responding to CSBV was investigated. In this study, Illumina high-throughput sequencing of Apis cerana larvae infected with CSBV for 24h and 48h were performed. DEmiRNAs and its target genes were analyzed by bioinformatics software, and GO functional annotation and KEGG pathway enrichment were analyzed. The sequencing results were validated by RT-qPCR. One hundred and forty known miRNAs and five novel miRNAs were identified in this study. A total of 18 DEmiRNAs were observed in L24I(the CSBV-infected group after 24 h) vs L24H(the health group of L24I) larva of A. cerana, including 12 upregulated and 6 downregulated miRNAs. 47DEmiRNAs were detected in L48I(the CSBV-infected group after 48 h) vs L48H(the health group of L48I), including 30 upregulated and 17 downregulated. Gene ontology analysis indicated some DEmiRNAs and related target genes were involved in functions including immune system processes and response to stimulus. KEGG pathway analysis showed that in addition to material and energy metabolism pathways, the main immune pathways include MAPK, lysosome, Hippo, and apoptosis signaling pathways. Validation by RT-qPCR showed that the expression trends of eight randomly selected miRNAs were consistent with the sequencing results, indicating accurate and reliable sequencing. This study not only helps us to deeply understand the changes of host miRNA during the process of CSBV infection, but also provides help to explore the function of miRNA during the interaction between Apis cerana and CSBV.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.004589
中图分类号:S895
引用信息:
[1]李一,肖宇婷,马跃宇,等.中华蜜蜂幼虫响应中蜂囊状幼虫病毒感染的MicroRNAs分析[J].病毒学报,2024,40(06):1366-1377.DOI:10.13242/j.cnki.bingduxuebao.004589.
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;国家自然科学基金（项目号：31772760）,题目：中华蜜蜂囊状幼虫病毒编码siRNA及其RNAi抑制因子在免疫保护中的作用; 辽宁省自然科学基金（项目号：2022-MS-385）,题目：CSBV非结构蛋白3C与宿主互作蛋白的筛选及其功能研究; 锦州医科大学横向课题（项目号：2022043）,题目：中国蜜蜂蜂群崩溃失调症流行情况调查研究~~
2024-09-06
2024-09-06
2024-09-06