SIGS vs HIGS: a study on the efficacy of two dsRNA delivery strategies to silence <i>Fusarium FgCYP51</i> genes in infected host and non‐host plants

Koch, Aline; Höfle, Lisa; Werner, B.; Imani, Jafargholi; Schmidt, Arthur H.; Jelonek, Lukas; Kögel, Karl-Heinz

doi:10.1111/mpp.12866

Cited by 56 publications

(60 citation statements)

References 51 publications

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“…Moreover, we found that the transferred SIGS-associated RNAs confer target gene silencing in the respective tissues as well as Fg disease resistance (Fig. 7, Koch et al, 2019). Interestingly, we measured the highest silencing efficiency in the tissue that was either directly sprayed or near to the sprayed locus.…”

Section: Discussionmentioning

confidence: 83%

“…In addition to the generation of RNA silencing signals in planta , plants can be protected from pathogens and pests by exogenously applied RNA biopesticides (known as spray-induced gene silencing, SIGS) (for review see: Mitter et al 2017; Cai et al 2018; Dubrovina and Kiselev 2019; Gaffar and Koch 2019; Dalakouras et al 2020). Over the last decade, our research (Koch et al, 2013; 2016; 2018; 2019; Höfle et al, 2019, Werner et al, 2019) and the work of others have revealed the enormous potential of RNA-silencing strategies as a potential alternative to conventional pesticides for plant protection, regardless of how target-specific inhibitory RNAs are applied (i.e. endogenously or exogenously).…”

Section: Introductionmentioning

confidence: 99%

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Phloem-mediated spreading of SIGS-derived non-coding RNAs inHordeum vulgare

Biedenkopf

Will

Knauer³

et al. 2019

Preprint

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1 7 Highlight 1 8The use of the SIGS-technology as a biopesticide will require the systemic spreading of 1 9 dsRNA/siRNA signals. Our findings strongly support the notion of phloem-mediated long-2 0 distance movement of SIGS-associated RNAs. Abstract 2 2 Small (s)RNA molecules are crucial factors in the communication between hosts and their 2 3 3 1 distance movement of SIGS-associated dsRNA and/or siRNA. These findings are significant 3 2 contributions to our mechanistic understanding of RNA spray technology, as our previous 3 3 data indicate that SIGS requires the processing of dsRNAs by the fungal RNAi machinery. In 3 4 summary, our findings support the model that SIGS involves: (i) uptake of sprayed dsRNA by 3 5 the plant (via stomata); (ii) transfer of apoplastic dsRNAs into the symplast (DCL processing 3 6 into siRNAs); (iii) systemic translocation of siRNA or unprocessed dsRNA via the vascular 3 7 system (phloem/xylem); (iv) uptake of apoplastic dsRNA or symplastic dsRNA/siRNA 3 8 depending on the lifestyle/feeding behavior of the pathogen/pest.3 9

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Section: Discussionmentioning

confidence: 83%

Section: Introductionmentioning

confidence: 99%

Phloem-mediated spreading of SIGS-derived non-coding RNAs inHordeum vulgare

Biedenkopf

Will

Knauer³

et al. 2019

Preprint

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“…Previously, we discovered that longer dsRNAs of 400–800 nt exhibited a higher gene-silencing efficiency and a stronger disease resistance than 200-nt dsRNAs (Koch et al, 2019), indicating that the quantity of siRNAs derived from a longer dsRNA precursor is simply higher. To test whether the length and/or the selected target gene sequence influences silencing efficiencies, we constructed 10 different dsRNA constructs targeting FgAGO/FgDCL pairs (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Regardless of how target-specific inhibitory RNAs are applied (i.e. endogenously or exogenously), the use of HIGS and SIGS technologies to control Fusarium species have been shown to be a potential alternative to conventional pesticides (Koch et al 2013; Ghag et al 2014; Cheng et al 2015; Hu et al 2015; Chen et al 2016; Pareek and Rajam 2017; Bharti et al 2017; Baldwin et al 2018; Koch et al 2018; Koch et al 2019), supporting the notion that RNAi strategies may improve food safety by controlling the growth of phytopathogenic, mycotoxin-producing fungi (reviewed by Machado et al 2017; Majumdar et al 2017).…”

Section: Introductionmentioning

confidence: 99%

“…In plants, RNAi strategies have the potential to protect host plants against predation or infection by pathogens and pests mediated by lethal RNAi signals generated in planta , a strategy known as ‘host-induced gene silencing’ (HIGS; Nowara et al 2010) (for review, see Koch and Kogel 2014; Yin and Hulbert 2015; Zhang et al, 2017; Qi et al, 2019; Gaffar and Koch 2019). In addition to the generation of RNA-silencing signals in planta , plants can be protected from pathogens and pests by spray-applied RNA biopesticides, known as spray-induced gene silencing (SIGS) (Koch et al, 2016; Wang et al, 2016; Konakalla et al, 2016; Mitter et al, 2017; Kaldis et al, 2018; Koch et al 2019). Regardless of how target-specific inhibitory RNAs are applied (i.e.…”

Section: Introductionmentioning

confidence: 99%

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RNA-spray-mediated silencing ofFusarium graminearum AGOandDCLgenes improve barley disease resistance

Werner

Gaffar

Schuemann

et al. 2019

Preprint

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AbstractOver the last decade, several studies have revealed the enormous potential of RNA-silencing strategies as a potential alternative to conventional pesticides for plant protection. We have previously shown that targeted gene silencing mediated by an in planta expression of non-coding inhibitory double-stranded RNAs (dsRNAs) can protect host plants against various diseases with unprecedented efficiency. In addition to the generation of RNA-silencing (RNAi) signals in planta, plants can be protected from pathogens and pests by spray-applied RNA-based biopesticides. Despite the striking efficiency of RNA-silencing-based technologies holds for agriculture, the molecular mechanisms underlying spray-induced gene silencing (SIGS) strategies are virtually unresolved, a requirement for successful future application in the field. Based on our previous work, we predict that the molecular mechanism of SIGS is controlled by the fungal-silencing machinery. In this study, we used SIGS to compare the silencing efficiencies of computationally-designed versus manually-designed dsRNA constructs targeting ARGONAUTE and DICER genes of Fusarium graminearum (Fg). We found that targeting key components of the fungal RNAi machinery via SIGS could protect barley leaves from Fg infection and that the manual design of dsRNAs resulted in higher gene-silencing efficiencies than the tool-based design. Moreover, our results indicate the possibility of cross-kingdom RNA silencing in the Fg-barley interaction, a phenomenon in which sRNAs operate as effector molecules to induce gene silencing between species from different kingdoms, such as a plant host and their interacting pathogens.

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Functionalized Carbon Dot‐Delivered RNA Nano Fungicides as Superior Tools to Control Phytophthora Pathogens through Plant RdRP1 Mediated Spray‐Induced Gene Silencing

Wang

Zhang

et al. 2023

Adv Funct Materials

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Spray-induced gene silencing (SIGS) represents an attractive avenue for plant protection, but limited uptake efficiency of double-stranded RNA (dsRNA) has restricted its application against the notorious oomycetes, Phytophthora. To control Phytophthora, dsRNAs targeting two essential genes in Phytophthora are screened for their ability to partially decrease Phytophthora capsici (P. capsici) infection and fecundity. To further facilitate SIGS for Phytophthora control, functionalized carbon dots (CDs) are complexed with the screened dsRNAs (dsRNA-CDs) through electrostatic interaction. dsRNA-CDs significantly enhanced the control efficacy of dsRNA against Phytophthora infestans, Phytophthora sojae, and both the wild-type and fungicide-resistant P. capsici. The synergism is based on enhancing dsRNA stability and internalization. Dual treatment with dsRNA-CDs and the corresponding fungicide reduced the amount of fungicide required to achieve the same protective effect by 90%. Plant RdRP1 is the main effector in processing various lengths of small RNAs to induce translation inhibition in Phytophthora. Notably, here the first application of a nano-delivery system to improve the effect of SIGS against Phytophthora pathogens is reported. Moreover, the elucidation of how CD facilitates dsRNA internalization in recipient cell and the molecular mechanism of SIGS can benefit the application of dsRNA-CDs in other plant-pathogen pathosystems through improving dsRNA bioactivity and reducing chemical fungicides application.

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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

Cited by 56 publications

References 51 publications

Phloem-mediated spreading of SIGS-derived non-coding RNAs inHordeum vulgare

Phloem-mediated spreading of SIGS-derived non-coding RNAs inHordeum vulgare

RNA-spray-mediated silencing ofFusarium graminearum AGOandDCLgenes improve barley disease resistance

Functionalized Carbon Dot‐Delivered RNA Nano Fungicides as Superior Tools to Control Phytophthora Pathogens through Plant RdRP1 Mediated Spray‐Induced Gene Silencing

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