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口服双链RNA可导致亚洲柑橘木虱若虫和成虫死亡。

Oral delivery of double-stranded RNAs induces mortality in nymphs and adults of the Asian citrus psyllid, Diaphorina citri.

作者信息

Galdeano Diogo Manzano, Breton Michèle Claire, Lopes João Roberto Spotti, Falk Bryce W, Machado Marcos Antonio

机构信息

Laboratório de Biotecnologia, Centro de Citricultura Sylvio Moreira, Instituto Agronômico de Campinas, Cordeirópolis, São Paulo, Brazil.

Instituto de Biologia, Universidade Estadual de Campinas, Campinas, São Paulo, Brazil.

出版信息

PLoS One. 2017 Mar 10;12(3):e0171847. doi: 10.1371/journal.pone.0171847. eCollection 2017.

Abstract

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama, is one of the most important citrus pests. ACP is the vector of the phloem-limited bacteria Candidatus Liberibacter americanus and Candidatus Liberibacter asiaticus, the causal agents of the devastating citrus disease huanglongbing (HLB). The management of HLB is based on the use of healthy young plants, eradication of infected plants and chemical control of the vector. RNA interference (RNAi) has proven to be a promising tool to control pests and explore gene functions. Recently, studies have reported that target mRNA knockdown in many insects can be induced through feeding with double-stranded RNA (dsRNA). In the current study, we targeted the cathepsin D, chitin synthase and inhibitor of apoptosis genes of adult and nymph ACP by feeding artificial diets mixed with dsRNAs and Murraya paniculata leaves placed in dsRNAs solutions, respectively. Adult ACP mortality was positively correlated with the amount of dsRNA used. Both nymphs and adult ACP fed dsRNAs exhibited significantly increased mortality over time compared with that of the controls. Moreover, qRT-PCR analysis confirmed the dsRNA-mediated RNAi effects on target mRNAs. These results showed that RNAi can be a powerful tool for gene function studies in ACP and perhaps for HLB control.

摘要

亚洲柑橘木虱(Diaphorina citri Kuwayama)是最重要的柑橘害虫之一。亚洲柑橘木虱是韧皮部限制性细菌美洲韧皮杆菌(Candidatus Liberibacter americanus)和亚洲韧皮杆菌(Candidatus Liberibacter asiaticus)的传播媒介,这两种细菌是毁灭性柑橘病害黄龙病(HLB)的致病因子。黄龙病的防治基于使用健康的幼树、清除受感染的植株以及对传播媒介进行化学防治。RNA干扰(RNAi)已被证明是控制害虫和探索基因功能的一种有前景的工具。最近,有研究报道通过喂食双链RNA(dsRNA)可诱导许多昆虫体内的靶标mRNA敲低。在本研究中,我们分别通过喂食混合dsRNA的人工饲料以及将九里香叶片置于dsRNA溶液中,针对成虫和若虫亚洲柑橘木虱的组织蛋白酶D、几丁质合成酶和凋亡抑制基因进行研究。成虫亚洲柑橘木虱的死亡率与所用dsRNA的量呈正相关。与对照相比,喂食dsRNA的若虫和成虫亚洲柑橘木虱随着时间的推移死亡率均显著增加。此外,qRT-PCR分析证实了dsRNA介导的对靶标mRNA的RNAi效应。这些结果表明,RNAi可能是亚洲柑橘木虱基因功能研究以及或许用于黄龙病防治的一种有力工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a033/5345766/83307d6cd9dd/pone.0171847.g001.jpg

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