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RNA 测序鉴定出一种卵巢特异性 microRNA,miR-311-3p,通过靶向 Bactrocera dorsalis 中的内收蛋白 B1 参与卵巢发育和生育力。

RNA sequencing identifies an ovary-enriched microRNA, miR-311-3p, involved in ovarian development and fecundity by targeting Endophilin B1 in Bactrocera dorsalis.

机构信息

Key Laboratory of Horticultural Plant Biology (MOE), Hubei Key Laboratory of Insect Resource Application and Sustainable Pest Control, Institute of Urban and Horticultural Entomology, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, China.

出版信息

Pest Manag Sci. 2023 Feb;79(2):688-700. doi: 10.1002/ps.7236. Epub 2022 Nov 18.

Abstract

BACKGROUND

The oriental fruit fly, Bactrocera dorsalis, is a highly invasive pest in East Asia and the Pacific. With the development of pesticides resistance, environment-friendly pesticides are urgently needed. MicroRNAs (miRNAs) are critical regulators of numerous biological processes, including reproduction. Thus, it is significant to identify reproductive-related miRNAs in this notorious pest to facilitate its control, such as RNAi-based biopesticides targeting essential miRNAs.

RESULTS

A high-throughput sequencing was carried out to identify miRNAs involved in reproduction from the ovary and fat body at four developmental stages [1 day (d), 5, 9, and 13 days post-eclosion] in female B. dorsalis. Results showed that 98 and 74 miRNAs were differentially expressed in ovary and fat body, respectively, during sexual maturation. Gene ontology analysis showed that target genes involved in oogenesis and lipid particle accounted for 33% and 15% of the total targets, respectively. Among these differentially expressed miRNAs, we found by qPCR that miR-311-3p was enriched in the ovary and down-regulated during sexual maturation. Injection of agomir-miR-311-3p resulted in arrested ovarian development, reduced egg deposition and progeny viability. Endophilin B1 was confirmed to be the target of miR-311-3p, via dual-luciferase assay and expression profiling. Knockdown of Endophilin B1 resulted in reproductive defects similar to those caused by injection of miR-311-3p agomir. Thus, miR-311-3p might play a critical role in female reproduction by targeting Endophilin B1.

CONCLUSION

Our data not only provides knowledge on the abundance of reproductive-related miRNAs and target genes, but also promotes new control strategies for this pest. © 2022 Society of Chemical Industry.

摘要

背景

东方果实蝇是一种高度入侵性害虫,分布于东亚和太平洋地区。随着农药抗性的发展,人们迫切需要环境友好型农药。微小 RNA(miRNA)是许多生物过程(包括生殖)的关键调控因子。因此,鉴定这种臭名昭著的害虫的生殖相关 miRNA 对于促进其防治具有重要意义,例如针对必需 miRNA 的 RNAi 为基础的生物农药。

结果

对性成熟过程中四个发育阶段(1 天(d)、5、9 和 13 天出蛹)的雌果蝇卵巢和脂肪体中的生殖相关 miRNA 进行了高通量测序。结果表明,卵巢和脂肪体中分别有 98 和 74 个 miRNA 存在差异表达。基因本体分析表明,卵发生和脂质颗粒相关的靶基因分别占总靶基因的 33%和 15%。在这些差异表达的 miRNA 中,我们通过 qPCR 发现 miR-311-3p 在卵巢中富集,并在性成熟过程中下调。注射 agomir-miR-311-3p 导致卵巢发育停滞、产卵减少和后代活力降低。双荧光素酶报告基因实验和表达谱分析证实内吞蛋白 B1 是 miR-311-3p 的靶基因。内吞蛋白 B1 敲低导致的生殖缺陷与注射 miR-311-3p agomir 引起的生殖缺陷相似。因此,miR-311-3p 通过靶向内吞蛋白 B1 可能在雌性生殖中发挥关键作用。

结论

本研究不仅为生殖相关 miRNA 和靶基因的丰度提供了知识,还为该害虫的防治提供了新的策略。© 2022 化学工业协会。

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