CYP3RNA, a double‐stranded (ds)RNA designed to concomitantly target the two sterol 14α‐demethylase genes FgCYP51A and FgCYP51B and the fungal virulence factor FgCYP51C, inhibits the growth of the ascomycete fungus Fusarium graminearum (Fg) in vitro and in planta. Here we compare two different methods (setups) of dsRNA delivery, viz. transgene expression (host‐induced gene silencing, HIGS) and spray application (spray‐induced gene silencing, SIGS), to assess the activity of CYP3RNA and novel dsRNA species designed to target one or two FgCYP51 genes. Using Arabidopsis and barley, we found that dsRNA designed to target two FgCYP51 genes inhibited fungal growth more efficiently than dsRNA targeting a single gene, although both dsRNA species reduced fungal infection. Either dsRNA delivery method reduced fungal growth stronger than anticipated from previous mutational knock‐out (KO) strategies, where single gene KO had no significant effect on fungal viability. Consistent with the strong inhibitory effects of the dsRNAs on fungal development in both setups, we detected to a large extent dsRNA‐mediated co‐silencing of respective non‐target FgCYP51 genes. Together, our data further support the valuation that dsRNA applications have an interesting potential for pesticide target validation and gene function studies, apart from their potential for crop protection.

中文翻译：

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SIGS 与 HIGS：关于两种 dsRNA 递送策略沉默受感染宿主和非宿主植物中镰刀菌 FgCYP51 基因的功效的研究。

CYP3RNA 是一种双链 (ds)RNA，旨在同时靶向两个甾醇 14α-去甲基酶基因FgCYP51A和FgCYP51B以及真菌毒力因子FgCYP51C ，在体外和植物中抑制子囊菌禾谷镰刀菌( Fg )的生长。在这里，我们比较了两种不同的 dsRNA 传递方法（设置），即。转基因表达（宿主诱导基因沉默，HIGS）和喷雾应用（喷雾诱导基因沉默，SIGS），以评估 CYP3RNA 和旨在靶向一两个FgCYP51基因的新型 dsRNA 物种的活性。使用拟南芥和大麦，我们发现针对两个FgCYP51基因设计的 dsRNA 比针对单个基因的 dsRNA 更有效地抑制真菌生长，尽管这两种 dsRNA 都能减少真菌感染。两种 dsRNA 递送方法对真菌生长的抑制作用均强于之前突变敲除 (KO) 策略的预期，其中单基因 KO 对真菌活力没有显着影响。与两种设置中 dsRNA 对真菌发育的强烈抑制作用一致，我们在很大程度上检测到 dsRNA 介导的各个非靶FgCYP51基因的共沉默。总之，我们的数据进一步支持了这样的评估：dsRNA 应用除了在作物保护方面的潜力外，在农药靶标验证和基因功能研究方面也具有令人感兴趣的潜力。