RNA Interference towards the Potato Psyllid, Bactericera cockerelli, Is Induced in Plants Infected with Recombinant Tobacco mosaic virus (TMV).
RNA Interference towards the Potato Psyllid, Bactericera cockerelli, Is Induced in Plants Infected with Recombinant Tobacco mosaic virus (TMV).
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DOI：
10.1371/journal.pone.0066050
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Falk BW
中科院分区：
文献类型：
--
作者：
Wuriyanghan H;Falk BW
马铃薯/番茄木虱Bactericera cockerelli（B. cockerelli）是一种重要的植物害虫，也是韧皮部限制性细菌青枯菌（Solanacearum）的载体，该细菌与马铃薯的斑马片病有关。以前，我们报道了在B中诱导RNA干扰效应。cockerelli通过胸内注射后体外制备的dsRNA/siRNA，以及饲喂含有这些效应RNA的人工饲料后。为了通过植物宿主传递RNAi效应子并快速鉴定取食植物的B中的有效靶序列。Cockerelli，在这里我们开发了一种基于植物病毒载体的植物内系统，用于评估候选序列。结果表明，含B. cockerelli序列可以在番茄（Solanumlycopersicum）、番茄（Physalisphiladelphica）和烟草（Nicotianatabacum）植物中有效地感染和产生小干扰RNA，更重要的是，通过TMV递送干扰序列在B中诱导RNAi效应，如通过肌动蛋白和V-ATP酶mRNA减少所测量的。以这些植物为食的小公鸡RNA干扰效应主要在B中检测到。鸡的内脏与我们用TMV的结果相反，重组马铃薯X病毒（PVX）和烟草脆裂病毒（TRV）没有在所有植物中产生稳健的感染，并且在B中没有诱导可检测的RNAi效应。cockerelli当B.使小公鸡以从相应的TMV感染植物的接种或下部展开的叶收集的叶圆片为食。用含有B的重组TMV感染番茄植株。cockerelli肌动蛋白或V-ATP酶序列也显示出导致B降低的表型效应。cockerelli后代产量与用含有GFP的重组TMV感染的植物相比。这些结果表明，可以在植物中实现针对韧皮部饲养者B的RNAi效应。cockerelli的方法和TMV-植物系统将为在植物中筛选合适的RNAi靶序列提供更快和更方便的方法。
The potato/tomato psyllid, Bactericera cockerelli (B. cockerelli), is an important plant pest and the vector of the phloem-limited bacterium Candidatus Liberibacter psyllaurous (solanacearum), which is associated with the zebra chip disease of potatoes. Previously, we reported induction of RNA interference effects in B. cockerelli via in vitro-prepared dsRNA/siRNAs after intrathoracic injection, and after feeding of artificial diets containing these effector RNAs. In order to deliver RNAi effectors via plant hosts and to rapidly identify effective target sequences in plant-feeding B. cockerelli, here we developed a plant virus vector-based in planta system for evaluating candidate sequences. We show that recombinant Tobacco mosaic virus (TMV) containing B. cockerelli sequences can efficiently infect and generate small interfering RNAs in tomato (Solanum lycopersicum), tomatillo (Physalis philadelphica) and tobacco (Nicotiana tabacum) plants, and more importantly delivery of interfering sequences via TMV induces RNAi effects, as measured by actin and V-ATPase mRNA reductions, in B. cockerelli feeding on these plants. RNAi effects were primarily detected in the B. cockerelli guts. In contrast to our results with TMV, recombinant Potato virus X (PVX) and Tobacco rattle virus (TRV) did not give robust infections in all plants and did not induce detectable RNAi effects in B. cockerelli. The greatest RNA interference effects were observed when B. cockerelli nymphs were allowed to feed on leaf discs collected from inoculated or lower expanded leaves from corresponding TMV-infected plants. Tomatillo plants infected with recombinant TMV containing B. cockerelli actin or V-ATPase sequences also showed phenotypic effects resulting in decreased B. cockerelli progeny production as compared to plants infected by recombinant TMV containing GFP. These results showed that RNAi effects can be achieved in plants against the phloem feeder, B. cockerelli, and the TMV-plant system will provide a faster and more convenient method for screening of suitable RNAi target sequences in planta.
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