过表达ace-miR-14-y对中华蜜蜂工蜂幼虫靶 基因表达及个体和肠道重量的影响
Overexpression of ace-miR-14-y affects target gene and gut weight of Apis cerana cerana larvae
徐国钧1** 刘小玉1** 刘治滩1 任亚萍1 康 婧1 宓诗雨1 胡艳雯1 刘彩珍2 陈大福1, 3, 4 郭 睿1, 3, 4***
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DOI：10.7679/j.issn.2095-1353.2025.011
作者单位：1. 福建农林大学蜂学与生物医药学院，福州 350002；2. 福建农业职业技术学院，福州 350002； 3. 天然生物毒素国家地方联合工程实验室, 福州 350002；4. 福建农林大学蜂疗研究所，福州 350002
中文关键词： 中华蜜蜂；幼虫；ace-miR-14-y；过表达；靶基因
英文关键词：Apis cerana cerana; larva; ace-miR-14-y; overexpression; target gene
中文摘要：
【目的】 探究过表达ace-miR-14-y对中华蜜蜂Apis cerana cerana工蜂幼虫靶基因表达及个体和肠道重量的影响，进而揭示ace-miR-14-y的潜在调控作用。【方法】 使用茎环RT-PCR和Sanger测序验证ace-miR-14-y的表达和序列。采用相关软件预测和分析ace-miR-14-y的靶基因。通过饲喂模拟物（Mimic）进行幼虫肠道中ace-miR-14-y的过表达。利用RT-qPCR检测ace-miR-14-y的过表达效果及过表达ace-miR-14-y后靶基因的相对表达量。使用电子天平对幼虫个体和肠道进行称重。【结果】 ace-miR-14-y在工蜂幼虫肠道中存在和表达；ace-miR-14-y共靶向265个基因，涉及33个GO条目与180条KEGG通路；，mimic-miR-14组ace-miR-14-y的表达量显著高于mimic-NC组（P<0.05）；过表达ace-miR-14-y后，靶基因Wg和AP-1的表达量均显著下调（P<0.05），幼虫个体和肠道的重量均极显著上升（P<0.01）。【结论】 过表达ace-miR-14-y显著影响中华蜜蜂工蜂幼虫肠道中靶基因Wg和AP-1的表达及幼虫个体和肠道的重量，ace-miR-14-y通过调节Wg和AP-1表达发挥潜在的调控作用。
英文摘要：
[Aim] The objective of this study is to
investigate the effect of ace-miR-14-y overexpression on expression of target
genes in the Apis cerana cerana worker larvae as well as weights of
individual and gut. [Methods] Stem-loop RT-PCR and Sanger
sequencing were utilized to confirm the expression and sequence of
ace-miR-14-y. Relative software were used to predict and analyze the target
genes of ace-miR-14-y. Overexpression
of ace-miR-14-y in the larval gut was performed by feeding mimic, followed by
examination of the effect of overexpression and the relative expression level
of target genes after overexpression. Electronic balance was employed to weigh
the larval individual and gut. [Results] The results demonstrated that ace-miR-14-y authentically
existed and expressed in the A. cerana cerana worker larval gut.
ace-miR-14-y targeted a total of 265 genes, involving 33 GO terms and 180 KEGG
pathways. The expression level of ace-miR-14-y in the mimic-miR-14 group was
significantly higher than that in the mimic-NC group (P<0.05). After
overexpression of ace-miR-14-y, the expression level of target genes Wg and AP-1 were both significantly downregulated (P<0.05) and
the weights of larval individual and gut were increased with extreme
significance (P<0.01). [Conclusion] The overexpression of ace-miR-14-y significantly affects the
target genes Wg and AP-1’s expression in Apis cerana cerana worker larval gut as well as the weights of larval individual and
gut, ace-miR-14-y plays a potential regulatory part through the regulation of
expression of Wg and AP-1.