Laboratory of Plant Breeding and Biometry, Faculty of Crop Science, Agricultural University of Athens, 11855, Athens, Greece.
Department of Agricultural Biotechnology, Assam Agricultural University, Jorhat, Assam, 785013, India.
Planta. 2017 Dec;246(6):1233-1241. doi: 10.1007/s00425-017-2776-7. Epub 2017 Sep 18.
Exogenously applied double-stranded RNA (dsRNA) molecules onto tomato leaves, moved rapidly from local to systemic leaves and were uptaken by agricultural pests namely aphids, whiteflies and mites. Four small interfering RNAs, deriving from the applied dsRNA, were molecularly detected in plants, aphids and mites but not in whiteflies. Double-stranded RNA (dsRNA) acts as the elicitor molecule of the RNA silencing (RNA interference, RNAi), the endogenous and evolutionary conserved surveillance system present in all eukaryotes. DsRNAs and their subsequent degradation products, namely the small interfering RNAs (siRNAs), act in a sequence-specific manner to control gene expression. Exogenous application of dsRNAs onto plants elicits resistance against plant viruses. In the present work, exogenously applied dsRNA molecules, derived from Zucchini yellow mosaic virus (ZYMV) HC-Pro region, onto tomato plants were detected in aphids (Myzus persicae), whiteflies (Trialeurodes vaporariorum) and mites (Tetranychus urticae) that were fed on treated as well as systemic tomato leaves. Furthermore, four siRNAs, deriving from the dsRNA applied, were detected in tomato and the agricultural pests fed on treated tomato plants. More specifically, dsRNA was detected in agricultural pests at 3 and 10 dpt (days post treatment) in dsRNA-treated leaves and at 14 dpt in systemic leaves. In addition, using stem-loop RT-PCR, siRNAs were detected in agricultural pests at 3 and 10 dpt in aphids and mites. Surprisingly, in whiteflies carrying the applied dsRNA, siRNAs were not molecularly detected. Our results showed that, upon exogenous application of dsRNAs molecules, these moved rapidly within tomato and were uptaken by agricultural pests fed on treated tomato. As a result, this non-transgenic method has the potential to control important crop pests via RNA silencing of vital genes of the respective pests.
将双链 RNA（dsRNA）分子施加到番茄叶片上，dsRNA 会迅速从局部叶片转移到系统叶片，并被农业害虫（如蚜虫、粉虱和螨虫）吸收。在植物、蚜虫和螨虫中检测到了源自施加的 dsRNA 的四种小干扰 RNA（siRNA），但在粉虱中没有检测到。dsRNA 作为 RNA 沉默（RNA 干扰，RNAi）的诱导分子，是所有真核生物中存在的内源性和进化上保守的监控系统。dsRNA 及其随后的降解产物，即小干扰 RNA（siRNA），以序列特异性的方式发挥作用来控制基因表达。将 dsRNA 施加到植物上可引发植物对病毒的抗性。在本工作中，将源自西葫芦黄花叶病毒（ZYMV）HC-Pro 区的 dsRNA 分子施加到番茄植株上，发现其可在取食处理过和系统叶片的蚜虫（桃蚜）、粉虱（烟粉虱）和螨虫（茶黄螨）中检测到。此外，还在取食处理过的番茄植株的番茄和农业害虫中检测到源自施加的 dsRNA 的四种 siRNA。更具体地说，在处理过的叶片中，dsRNA 在 3 和 10 dpt（处理后天数）时在农业害虫中检测到，而在系统叶片中在 14 dpt 时检测到。此外，使用茎环 RT-PCR，在取食处理过的番茄的蚜虫和螨虫中，在 3 和 10 dpt 时检测到 siRNA。令人惊讶的是，在携带施加的 dsRNA 的粉虱中，没有分子检测到 siRNA。我们的结果表明，dsRNA 分子施加后，它们在番茄体内迅速移动，并被取食处理过的番茄的农业害虫吸收。因此，这种非转基因方法有可能通过 RNA 沉默来控制各自害虫的重要作物害虫的关键基因。
