成体dsRNA喂养可同时沉默肠道核酸酶和一个重要的靶基因,从而提高RNAi效率,并增加毛角蝠成体的死亡率

Gennaro Volpe, Sarah Maria Mazzucchiello, Noemi Rosati, Francesca Lucibelli, Marianna Varone, Dora Baccaro, Ilaria Mattei, Ilaria Di Lelio, Andrea Becchimanzi, Ennio Giordano, Marco Salvemini, Serena Aceto, Francesco Pennacchio, Giuseppe Saccone
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引用次数: 0

摘要

地中海果蝇(Ceratitis capitata)是一种严重影响水果和蔬菜种植的主要双翅目害虫。目前,对它的控制主要依靠化学杀虫剂,但这些杀虫剂会带来健康风险,并对授粉昆虫造成影响。一种友好且针对特定物种的替代策略是通过喂食提供双链 RNA(dsRNA)来破坏害虫的基本功能,但这种方法在双翅目昆虫中的应用还很少。之前在直翅目和鞘翅目物种中的报道表明,dsRNA 在肠腔中被两种特异性核酸酶降解是昆虫摄食介导的 RNAi 的主要障碍之一。在我们的研究中,我们使用针对 ATPase 重要基因表达的 dsRNA 分子和两种肠道 dsRNA 核酸酶进行了为期三天的成虫喂养实验。这些dsRNA分子最近分别在两个Tephritidae物种中进行了测试,结果显示效果有限[1,2]。相比之下,我们在七天内观察到 79% 的死亡率,这与三个目标基因的 mRNA 水平下降有关。正如预期的那样,我们还观察到针对核酸酶的 RNAi 使 dsRNA 降解减少。这项研究说明了利用分子作为杀虫剂的潜力,通过靶向关键基因和肠道核酸酶来提高褐飞虱成虫的死亡率。此外,它还强调了探索基于 RNAi 的害虫管理方法的重要性。
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Simultaneous silencing of gut nucleases and a vital target gene by adult dsRNA feeding enhances RNAi efficiency and mortality in Ceratitis capitata adults
Ceratitis capitata, known as Mediterranean fruit fly (Medfly), is a major dipteran pest significantly impacting fruit and vegetable farming. Currently, its control heavily relies mainly on chemical insecticides, which pose health risks and have effects on pollinators. A friendly and species-specific alternative strategy involves providing double-stranded RNA (dsRNA) through feeding to disrupt essential functions in pest insects, which is poorly explored in dipteran species. Previous reports in Orthoptera and Coleoptera species suggested that dsRNA degradation by two specific nucleases in the intestinal lumen is among the major obstacle to feeding-mediated RNAi in insects. In our study, we experimented with three-day adult feeding using a combination of dsRNA molecules that target the expression of the ATPase vital gene and two intestinal dsRNA nucleases. These dsRNA molecules were recently tested separately in two Tephritidae species, showing limited effectiveness [1,2]. In contrast, we observed 79% mortality over seven days, which was associated with a decrease in mRNA levels of the three targeted genes. As expected, we also observed a reduction in dsRNA degradation following RNAi against nucleases. This research illustrates the potential of utilizing molecules as pesticides to achieve mortality rates in Medfly adults by targeting crucial genes and intestinal nucleases. Furthermore, it underscores the importance of exploring RNAi-based approaches for pest management
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