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Down-regulation of Fusarium oxysporum endogenous genes by Host-Delivered RNA interference enhances disease resistance

机译:宿主传递的RNA干扰下镰刀菌内源基因的下调增强了抗病能力

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Fusarium oxysporum is a devastating pathogen causing extensive yield losses in a variety of crops and development of sustainable, environmentally friendly methods to improve crop resistance is crucial. We have used Host-Derived RNA interference (HD-RNAi) technology to partially silence three different genes (FOW2, FRP1 and OPR) in the hemi-biotrophic fungus Fusarium oxysporum f. sp. conglutinans. Expression of double stranded RNA molecules targeting fungal pathogen genes was achieved in a number of transgenic Arabidopsis lines. F. oxysporum infecting the transgenic lines displayed substantially reduced mRNA levels on all three targeted genes, with an average of 75%, 83% and 72% reduction for FOW2, FRP1 and OPR respectively. The silencing of pathogen genes had a clear positive effect on the ability of the transgenic lines to fight infection. All transgenic lines displayed enhanced resistance to F. oxysporum with delayed disease symptom development, especially FRP1 and OPR lines. Survival rates after fungal infection were higher in the transgenic lines compared to control wild type plants which consistently showed survival rates of 10%, with FOW2 lines showing 25% survival; FRP1 lines 30-50% survival and FOW2 between 45-70% survival. The down-regulation effect was specific for the targeted genes without unintended effects in related genes. In addition to producing resistant crops, HD-RNAi can provide a useful tool to rapidly screen candidate fungal pathogenicity genes without the need to produce fungal knockout mutants.
机译:尖孢镰刀菌(Fusarium oxysporum)是毁灭性的病原体,其导致多种作物的大量产量损失,因此开发可持续的,环境友好的方法以提高作物的抗性至关重要。我们已使用宿主衍生的RNA干扰(HD-RNAi)技术部分沉默了半生营养真菌尖孢镰刀菌f中的三个不同基因(FOW2,FRP1和OPR)。 sp。粘蛋白。在许多转基因拟南芥品系中实现了靶向真菌病原体基因的双链RNA分子的表达。侵染转基因品系的尖孢镰刀菌在所有三个靶向基因上均显示出显着降低的mRNA水平,FOW2,FRP1和OPR分别平均降低了75%,83%和72%。病原体基因的沉默对转基因品系抵抗感染的能力具有明显的积极影响。所有转基因品系均表现出增强的对尖孢镰刀菌的抗性并延缓了疾病症状的发展,特别是FRP1和OPR品系。与对照野生型植物相比,转基因品系中真菌感染后的存活率更高,后者始终显示出10%的存活率,而FOW2品系显示出25%的存活率。 FRP1的存活率为30-50%,FOW2的存活率为45-70%。下调作用对于靶基因是特异性的,而在相关基因中没有意外作用。除了生产抗性农作物外,HD-RNAi还可以提供有用的工具来快速筛选候选真菌致病基因,而无需产生真菌敲除突变体。

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