Silencing of aphid genes by dsRNA feeding from plants.
Silencing of aphid genes by dsRNA feeding from plants.
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DOI：
10.1371/journal.pone.0025709
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Hogenhout SA
中科院分区：
文献类型：
--
作者：
Pitino M;Coleman AD;Maffei ME;Ridout CJ;Hogenhout SA
RNA干扰（RNAi）是研究包括蚜虫等半翅目昆虫在内的多种生物基因功能的重要反向遗传学工具。先前的工作已经表明，RNAi介导的豌豆蚜（Acyrthosiphon pisum）基因的敲除可以通过将双链RNA（dsRNA）或小干扰RNA（siRNA）直接注射到豌豆蚜血淋巴中或通过用含有小RNA的人工饲料喂养这些昆虫来实现。在这项研究中，我们已经开发了植物介导的RNAi技术的蚜虫，允许在蚜虫的自然环境中的基因沉默，并尽量减少这些昆虫在实验过程中的处理。绿色桃蚜M.选择桃蚜是因为它具有广泛的植物宿主范围，包括模式植物本氏烟草（Nicotianabenthamiana）和拟南芥（Arabidopsis thaliana），对于它们可以相对快速地产生转基因材料。我们瞄准了M。persicae Rack 1主要在肠道中表达; persicae C 002（MpC 002），主要在唾液腺中表达。以N. benthamiana叶盘瞬时产生对应于这些基因的dsRNA，并且在A.稳定产生dsRNA的拟南芥植物。MpC 002和Rack-1在转基因N. benthamiana和A. thaliana.此外，沉默的M。与这些具有基本功能的基因一致，桃产生较少的后代。在我们的植物介导的RNAi方法和公开的蚜虫沉默方法中实现了相似水平的基因沉默。此外，N.本氏叶盘试验可以发展为筛选以评估哪些基因是蚜虫在植物上存活所必需的。我们的研究结果也证明了植物介导的RNAi方法用于蚜虫控制的可行性。
RNA interference (RNAi) is a valuable reverse genetics tool to study gene function in various organisms, including hemipteran insects such as aphids. Previous work has shown that RNAi-mediated knockdown of pea aphid (Acyrthosiphon pisum) genes can be achieved through direct injection of double-stranded RNA (dsRNA) or small-interfering RNAs (siRNA) into the pea aphid hemolymph or by feeding these insects on artificial diets containing the small RNAs. In this study, we have developed the plant-mediated RNAi technology for aphids to allow for gene silencing in the aphid natural environment and minimize handling of these insects during experiments. The green peach aphid M. persicae was selected because it has a broad plant host range that includes the model plants Nicotiana benthamiana and Arabidopsis thaliana for which transgenic materials can relatively quickly be generated. We targeted M. persicae Rack1, which is predominantly expressed in the gut, and M. persicae C002 (MpC002), which is predominantly expressed in the salivary glands. The aphids were fed on N. benthamiana leaf disks transiently producing dsRNA corresponding to these genes and on A. thaliana plants stably producing the dsRNAs. MpC002 and Rack-1 expression were knocked down by up to 60% on transgenic N. benthamiana and A. thaliana. Moreover, silenced M. persicae produced less progeny consistent with these genes having essential functions. Similar levels of gene silencing were achieved in our plant-mediated RNAi approach and published silencing methods for aphids. Furthermore, the N. benthamiana leaf disk assay can be developed into a screen to assess which genes are essential for aphid survival on plants. Our results also demonstrate the feasibility of the plant-mediated RNAi approach for aphid control.
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影响因子：
11.4
作者：
Elbashir, SM;Martinez, J;Tuschl, T
通讯作者：
Tuschl, T
RNAi在豌豆蚜虫acyrthosiphon pisum中敲除基因。
影响因子：
3.5
作者：
Jaubert-Possamai, Stephanie;Le Trionnaire, Gael;Bonhomme, Joel;Christophides, Georges K;Rispe, Claude;Tagu, Denis
通讯作者：
Tagu, Denis
影响因子：
4.8
作者：
Liliental, J;Chang, DD
通讯作者：
Chang, DD
影响因子：
2.5
作者：
Kadrmas, Julie L.;Smith, Mark A.;Beckerle, Mary C.
通讯作者：
Beckerle, Mary C.
影响因子：
7.4
作者：
Johansen, LK;Carrington, JC
通讯作者：
Carrington, JC