Department of Phytopathology, University of Hohenheim, Stuttgart, Germany.
Methods Mol Biol. 2022;2523:79-91. doi: 10.1007/978-1-0716-2449-4_6.
To obtain direct evidence for the influence of an effector on the virulence or pathogenicity of a pathogen, it is necessary to knock out, knock down, or silence the respective gene. Since genetic transformation is not yet possible for rust fungi, silencing the gene is the only option. Posttranscriptional gene silencing uses RNAi. RNAi in plant pathogens can be accomplished by introducing dsRNA either by direct application of in vitro synthesized dsRNA or through positive-strand or double-strand RNA plant viruses. For studying effectors in Phakopsora pachyrhizi, we have implemented a host-induced silencing procedure based on virus-induced gene silencing using the bean pod mottle virus system. Here, procedures and interpretations of results are described and limitations of the system are discussed.
为了获得效应子对病原体毒力或致病性影响的直接证据,有必要敲除、敲低或沉默相应的基因。由于锈菌的遗传转化尚未实现,因此沉默基因是唯一的选择。转录后基因沉默利用 RNAi。植物病原体中的 RNAi 可以通过直接应用体外合成的 dsRNA 或通过正链或双链 RNA 植物病毒来实现。为了研究褐斑病菌中的效应子,我们已经实施了一种基于病毒诱导基因沉默的寄主诱导沉默程序,该程序使用菜豆荚斑驳病毒系统。在此,描述了程序和结果的解释,并讨论了该系统的局限性。