HvStaufenC contributes to the high RNAi efficiency in the 28-spotted ladybeetle, Henosepilachna vigintioctopunctata

IF 3.2 2区 农林科学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Zhaoyang Li , June-Sun Yoon , Zexin Zhong , Yalin Ruan , Chunxiao Yang , Xuguo Zhou , Youjun Zhang , Huipeng Pan
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Abstract

RNA interference (RNAi) has been shown to be relatively effective in coleopteran insects, with limited exploration into the molecular mechanisms that underlie this effectiveness. This study specifically examines the 28-spotted ladybeetle, Henosepilachna vigintioctopunctata (Hvig), known for its high RNAi efficiency. Here, we utilized RNAi and CRISPR/Cas9 techniques to identify and validate the genes involved in the RNAi pathway that enhance RNAi efficacy in Hvig. We identified a total of 15 potential genes within the RNAi pathway that may impact RNAi efficiency. The bioassay results showed that only knockdown of HvStaufenC in the 3rd instar larvae could block the abnormal body color phenotype and lethality induced by the subsequent silencing of the two marker genes, HvTH (tyrosine hydroxylase) and HvABCH1 (ATP-binding cassette H transporter gene), respectively. Additionally, successful CRISPR/Cas9-mediated knockout of HvStaufenC led to the generation of stable, heritable mutants that exhibited insensitivity to RNAi, displaying no response to RNAi targeting HvTH and HvABCH1. Compared to the wild-type strain, the HvStaufenC knockout (HvStaufenCKO) mutant females demonstrated a 42 % decrease in oviposition rate and a 41.3 % reduction in egg hatchability. This study demonstrates that HvStaufenC gene is crucial for the RNAi efficiency of Hvig and offers new evidence into the RNAi mechanisms in coleopteran species.

Abstract Image

HvStaufenC有助于28斑瓢虫(Henosepilachna vigintioctopunctata)的高RNAi效率
RNA干扰(RNAi)已被证明在鞘翅目昆虫中是相对有效的,但对这种有效性背后的分子机制的探索有限。这项研究特别研究了28斑点瓢虫,Henosepilachna vigintioctopunctata (Hvig),以其高RNAi效率而闻名。在这里,我们利用RNAi和CRISPR/Cas9技术来鉴定和验证RNAi途径中参与增强RNAi在Hvig疗效的基因。我们在RNAi途径中共鉴定了15个可能影响RNAi效率的潜在基因。生物实验结果表明,只有敲低HvStaufenC才能阻断3龄幼虫酪氨酸羟化酶(HvTH)和atp结合盒H转运体基因(HvABCH1)随后沉默导致的异常体色表型和致死率。此外,CRISPR/ cas9介导的HvStaufenC成功敲除导致产生稳定的、可遗传的突变体,这些突变体对RNAi不敏感,对靶向HvTH和HvABCH1的RNAi没有反应。与野生型菌株相比,HvStaufenCKO基因敲除突变雌性的产卵率降低了42%,卵子孵化率降低了41.3%。本研究表明,HvStaufenC基因对Hvig的RNAi效率起着至关重要的作用,为研究鞘翅类物种的RNAi机制提供了新的证据。
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来源期刊
CiteScore
7.40
自引率
5.30%
发文量
105
审稿时长
40 days
期刊介绍: This international journal publishes original contributions and mini-reviews in the fields of insect biochemistry and insect molecular biology. Main areas of interest are neurochemistry, hormone and pheromone biochemistry, enzymes and metabolism, hormone action and gene regulation, gene characterization and structure, pharmacology, immunology and cell and tissue culture. Papers on the biochemistry and molecular biology of other groups of arthropods are published if of general interest to the readership. Technique papers will be considered for publication if they significantly advance the field of insect biochemistry and molecular biology in the opinion of the Editors and Editorial Board.
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