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Effective and specific in planta RNAi in cyst nematodes: expression interference of four parasitism genes reduces parasitic success

机译:在囊肿线虫的植物RNAi中有效且特异性:四种寄生基因的表达干扰减少了寄生虫的成功

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摘要

Cyst nematodes are highly evolved sedentary plant endoparasites that use parasitism proteins injected through the stylet into host tissues to successfully parasitize plants. These secretory proteins likely are essential for parasitism as they are involved in a variety of parasitic events leading to the establishment of specialized feeding cells required by the nematode to obtain nourishment. With the advent of RNA interference (RNAi) technology and the demonstration of host-induced gene silencing in parasites, a new strategy to control pests and pathogens has become available, particularly in root-knot nematodes. Plant host-induced silencing of cyst nematode genes so far has had only limited success but similarly should disrupt the parasitic cycle and render the host plant resistant. Additional in planta RNAi data for cyst nematodes are being provided by targeting four parasitism genes through host-induced RNAi gene silencing in transgenic Arabidopsis thaliana, which is a host for the sugar beet cyst nematode Heterodera schachtii. Here it is reported that mRNA abundances of targeted nematode genes were specifically reduced in nematodes feeding on plants expressing corresponding RNAi constructs. Furthermore, this host-induced RNAi of all four nematode parasitism genes led to a reduction in the number of mature nematode females. Although no complete resistance was observed, the reduction of developing females ranged from 23% to 64% in different RNAi lines. These observations demonstrate the relevance of the targeted parasitism genes during the nematode life cycle and, potentially more importantly, suggest that a viable level of resistance in crop plants may be accomplished in the future using this technology against cyst nematodes.
机译:囊肿线虫是高度进化的久坐植物内寄生物,它使用通过管心针注入宿主组织的寄生蛋白成功寄生植物。这些分泌蛋白可能是寄生虫必不可少的,因为它们参与各种寄生虫事件,导致建立线虫获取营养所需的专门饲养细胞。随着RNA干扰(RNAi)技术的出现以及寄生虫中宿主诱导的基因沉默的证明,一种控制病虫害和病原体的新策略已经问世,尤其是在根结线虫中。迄今为止,植物宿主诱导的囊肿线虫基因沉默仅取得了有限的成功,但同样应该破坏寄生周期并赋予宿主植物抗性。通过在转基因拟南芥中的宿主诱导的RNAi基因沉默,通过靶向四个寄生虫基因来提供囊肿线虫的其他植物RNAi数据,该基因是甜菜囊肿线虫Heterodera schachtii的宿主。据报道,在以表达相应RNAi构建体的植物为食的线虫中,靶向线虫基因的mRNA丰度特别降低。此外,所有四个线虫寄生基因的宿主诱导的RNAi导致成熟线虫雌性数量的减少。尽管未观察到完全抗药性,但在不同的RNAi品系中,发育中的雌性的减少范围为23%至64%。这些观察结果证明了在线虫生命周期中靶向的寄生虫基因的相关性,并且可能更重要的是,表明使用该技术针对囊肿线虫的抗病性可能在将来实现。

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