遵义医科大学学报2025，Vol.48Issue(5)：423-436,14.
基于转录组测序研究眼斑芫菁幼虫体内斑蝥素生物合成的调控机制
Study on the regulatory mechanism of cantharidin biosynthesis in the Larvae of Mylabris cichorii Linnaeus based on transcriptome sequencing
摘要
Abstract
Objective This study employed transcriptome sequencing and molecular biological techniques to in-vestigate the regulatory mechanisms underlying cantharidin biosynthesis during the larval stage of M.cichorii.Methods Laboratory populations of M.cichorii were established,and larvae at 1-5 instars were collected for tran-scriptome sequencing with three biological replicates.Bioinformatics analysis was conducted to identify target genes involved in the regulation of cantharidin biosynthesis.Phylogenetic trees were constructed,and the identi-fied target genes were cloned and systematically named.RNA interference(RNAi)technology was employed to preliminarily validate the regulatory mechanism of the target gene,the interference effect was detected by RT-qPCR technology,and the content of cantharidin was measured by GC-MS.Results Transcriptome sequencing a-nalysis identified only one key gene.Gene structure analysis and phylogenetic tree construction revealed that the target gene belongs to the NPC1b subfamily.Further cloning,sequencing,and alignment of the target gene led to its designation as McicNPC1b.The coding sequence(CDS)of McicNPC1b is 3 729 bp in length.RNA inter-ference(RNAi)results showed that downregulation of McicNPC1b expression led to an extremely significant re-duction in cantharidin levels in larvae(P＜0.01).RT-qPCR results indicated that the expression pattern of McicNPC1b across larval instars slightly differed from the trend of cantharidin biosynthesis.The expression peak of McicNPC1b occurred during the 4th instar.Further research found that when McicNPC1b expression was downregulated,the expression of MFE,an upstream gene in the cantharidin biosynthesis pathway,was signifi-cantly upregulated(P＜0.05).Conclusion McicNPC1b influences the biosynthesis of cantharidin,and its mech-anism may involve the regulation of cantharidin biosynthesis by inversely modulating the secretion of juvenile hor-mone(JH),which is a precursor substance for cantharidin synthesis.关键词
眼斑芜菁/斑蝥素/保幼激素/调控机制Key words
Mylabris cichorii Linnaeus/cantharidin/juvenile hormone/regulatory mechanism分类
中医学引用本文复制引用
张文,杨西京,袁孟黔,张建永,李萨丽,李晓飞..基于转录组测序研究眼斑芫菁幼虫体内斑蝥素生物合成的调控机制[J].遵义医科大学学报,2025,48(5):423-436,14.基金项目
国家自然科学基金青年基金项目(NO:82404794) （NO:82404794）
贵州省教育厅青年科技人才成长项目[NO:黔教技(2024)-130]. （2024）
