SIGS vs HIGS: a study on the efficacy of two dsRNA delivery strategies to silence Fusarium FgCYP51 genes in infected host and non-host plants
SIGS vs HIGS: a study on the efficacy of two dsRNA delivery strategies to silence Fusarium FgCYP51 genes in infected host and non-host plants
复制标题
DOI：
10.1111/mpp.12866
复制
发表时间：
2019-10-11
影响因子：
4.9
通讯作者：
Kogel, Karl-Heinz
中科院分区：
文献类型：
--
作者：
Koch, Aline;Hoefle, Lisa;Kogel, Karl-Heinz
CYP 3RNA是一种双链（ds）RNA，其被设计为同时靶向两种甾醇14 α-脱甲基酶基因FgCYP 51 A和FgCYP 51 B以及真菌毒力因子FgCYP 51 C，在体外和植物中抑制子囊菌真菌禾谷镰刀菌（Fg）的生长。在这里，我们比较了两种不同的dsRNA递送方法（设置），即转基因表达（宿主诱导的基因沉默，HIGS）和喷雾应用（喷雾诱导的基因沉默，SIGS），以评估CYP 3RNA和设计用于靶向一个或两个FgCYP 51基因的新型dsRNA种类的活性。使用拟南芥和大麦，我们发现设计为靶向两个FgCYP 51基因的dsRNA比靶向单个基因的dsRNA更有效地抑制真菌生长，尽管两种dsRNA种类都减少了真菌感染。任一种dsRNA递送方法降低真菌生长的能力强于先前突变敲除（KO）策略的预期，其中单基因KO对真菌活力没有显著影响。与dsRNA在两种设置中对真菌发育的强抑制作用一致，我们在很大程度上检测到dsRNA介导的相应非靶FgCYP 51基因的共沉默。总之，我们的数据进一步支持了这样的评价，即dsRNA应用除了具有作物保护的潜力外，还具有农药靶标验证和基因功能研究的有趣潜力。
CYP3RNA, a double-stranded (ds)RNA designed to concomitantly target the two sterol 14 alpha-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.