scholarly journals Study on the efficacy of dsRNAs with increasing length in RNAi-based silencing of the Fusarium CYP51 genes

2019 ◽  
Author(s):  
L Höfle ◽  
A Shrestha ◽  
B Werner ◽  
L Jelonek ◽  
A Koch
Keyword(s):  
Disease Resistance ◽  
Gene Silencing ◽  
Fungal Infection ◽  
Disease Control ◽  
Fusarium Graminearum ◽  
Rna Silencing ◽  
Transgenic Arabidopsis ◽  
Plant Protection ◽  
Field Application ◽  
Rnai Technology

AbstractPreviously, we have demonstrated that transgenic Arabidopsis and barley plants, expressing a 791 nucleotide (nt) dsRNA (CYP3RNA) that targets all three CYP51 genes (FgCYP51A, FgCYP51B, FgCYP51C) in Fusarium graminearum (Fg), inhibited fungal infection via a process designated as host-induced gene silencing (HIGS). More recently, we have shown that spray applications of CYP3RNA also protect barley from fungal infection via a process termed spray-induced gene silencing (SIGS). Thus, RNAi technology may have the potential to revolutionize plant protection in agriculture. Therefore, successful field application will require optimization of RNAi design necessary to maximize the efficacy of the RNA silencing construct for making RNAi-based strategies a realistic and sustainable approach.Previous studies indicate that silencing is correlated with the number of siRNAs generated from a dsRNA precursor. To prove the hypothesis that silencing efficiency is correlated with the number of siRNAs processed out of the dsRNA precursor, we tested in a HIGS and SIGS approach dsRNA precursors of increasing length ranging from 400 nt to 1500 nt to assess gene silencing efficiency of individual FgCYP51 genes. Concerning HIGS-mediated disease control, we found that there is no significant correlation between the length of the dsRNA precursor and the reduction of Fg infection on CYP51-dsRNA expressing Arabidopsis plants. Importantly and in clear contrast to HIGS, we measured a decrease in SIGS-mediated Fg disease resistance that significantly correlates with the length of the dsRNA construct that was sprayed, indicating that the size of the dsRNA interferes with a sufficient uptake of dsRNAs by the fungus.

scholarly journals RNA-spray-mediated silencing of Fusarium graminearum AGO and DCL genes improve barley disease resistance

2019 ◽  
Author(s):  
B Werner ◽  
FY Gaffar ◽  
J Schuemann ◽  
D Biedenkopf ◽  
A Koch
Keyword(s):  
Gene Silencing ◽  
Fusarium Graminearum ◽  
Rna Silencing ◽  
Molecular Mechanisms ◽  
Plant Protection ◽  
Future Application ◽  
In Planta ◽  
Plant Host ◽  
Barley Leaves ◽  
Potential Alternative

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.

scholarly journals Detoxification and Excretion of Trichothecenes in Transgenic Arabidopsisthaliana Expressing Fusarium graminearum Trichothecene 3-O-acetyltransferase

Toxins ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 320
Author(s):  
Guixia Hao ◽  
Susan McCormick ◽  
Helene Tiley ◽  
Thomas Usgaard
Keyword(s):  
Fusarium Graminearum ◽  
Transgenic Arabidopsis ◽  
Plant Protection ◽  
Plant Cells ◽  
Wild Type ◽  
Head Blight ◽  
Protection Mechanism ◽  
The Media ◽  
Self Protection

Fusarium graminearum, the causal agent of Fusarium head blight (FHB), produces trichothecenes including deoxynivalenol (DON), nivalenol (NIV), and 3,7,15-trihydroxy-12,13-epoxytrichothec-9-ene (NX-3). These toxins contaminate grains and cause profound health problems in humans and animals. To explore exploiting a fungal self-protection mechanism in plants, we examined the ability of F. graminearum trichothecene 3-O-acetyltransferase (FgTri101) to detoxify several key trichothecenes produced by F. graminearum: DON, 15-ADON, NX-3, and NIV. FgTri101 was cloned from F. graminearum and expressed in Arabidopsis plants. We compared the phytotoxic effects of purified DON, NIV, and NX-3 on the root growth of transgenic Arabidopsis expressing FgTri101. Compared to wild type and GUS controls, FgTri101 transgenic Arabidopsis plants displayed significantly longer root length on media containing DON and NX-3. Furthermore, we confirmed that the FgTri101 transgenic plants acetylated DON to 3-ADON, 15-ADON to 3,15-diADON, and NX-3 to NX-2, but did not acetylate NIV. Approximately 90% of the converted toxins were excreted into the media. Our study indicates that transgenic Arabidopsis expressing FgTri101 can provide plant protection by detoxifying trichothecenes and excreting the acetylated toxins out of plant cells. Characterization of plant transporters involved in trichothecene efflux will provide novel targets to reduce FHB and mycotoxin contamination in economically important plant crops.

scholarly journals Селекция растений на устойчивость – основа защиты от болезней в органическом земледелии

2019 ◽  
Author(s):  
S.G. Monakhos ◽  
A.V. Voronina ◽  
A.V. Baidina ◽  
O.N. Zubko
Keyword(s):  
Quality Of Life ◽  
Disease Resistance ◽  
Organic Farming ◽  
Plant Protection ◽  
High Specificity ◽  
The One ◽  
Durable Disease Resistance

Одна из ключевых проблем, сдерживающих распространение органических технологий, – экологически безопасная защита растений от болезней и вредителей. В силу выраженной специфики органического земледелия (требования к использованию природоподобных технологий и запрета на использование пестицидов, надежная генетическая устойчивость сортов и гибридов становится одним из основных путей решения этой проблемы. Представлен аналитический обзор современных данных по этой тематике.The role and importance of organic farming in improving the quality of life of humanity are obvious. The one of the key problems limiting the organic technologies is the problem of plant protection against diseases and pests. Due to high specificity of organic farming technologies, particularly the requirements for the use of nature-like technologies and the restrictions on the use of pesticides, durable disease resistance of crops becomes super-actual.

The small RNA‐mediated gene silencing machinery is required in Arabidopsis for stimulation of growth, systemic disease resistance, and the suppression of nitrile‐specifier, NSP4 gene by Trichoderma atroviride

2021 ◽  
Author(s):  
Oscar Guillermo Rebolledo Prudencio ◽  
Magnolia Estrada Rivera ◽  
Mitzuko Dautt Castro ◽  
Mario A. Arteaga‐Vazquez ◽  
Catalina Arenas‐Huertero ◽  
...  
Keyword(s):  
Disease Resistance ◽  
Gene Silencing ◽  
Small Rna ◽  
Systemic Disease ◽  
Nsp4 Gene ◽  
Trichoderma Atroviride ◽  
Stimulation Of

Regulatory aspects of RNAi in plant production.

2021 ◽  
pp. 154-158
Author(s):  
Werner Schenkel ◽  
Achim Gathmann
Keyword(s):  
Small Rna ◽  
Plant Protection ◽  
Plant Protection Products ◽  
Plant Production ◽  
The European Union ◽  
Relevant Aspect ◽  
Rna Molecules ◽  
Rnai Technology ◽  
Living Organisms ◽  
Intended Use

Abstract Technologies based on RNA interference (RNAi) may be used in plant production in different contexts. With respect to applicable regulations, a major distinction is to be made between plants producing small RNA molecules due to modifications of the genome and topically applied plant protection products (PPPs) based on double-stranded RNA (dsRNA). The first group may be further divided into those using RNAi technology to achieve changes in the plant's metabolism and those where plant-produced RNA molecules are intended to impact other organisms that interact with the plant. For PPPs, relevant aspects are whether the product contains living organisms or only purified molecules. The intended use of the product is another relevant aspect with respect to regulation. It is expected that PPPs will be among the first products utilizing the RNAi mechanism in the European Union. This chapter discusses the regulation of modified RNAi plants and the regulation of PPPs utilizing RNAi mechanisms.

The future is now: revolution of RNA-mediated gene silencing in plant protection against insect pests and diseases

2020 ◽  
Vol 14 (6) ◽  
pp. 643-662
Author(s):  
Fook-Hwa Lim ◽  
Omar Abd Rasid ◽  
Abdul Wahab Mohd As’wad ◽  
Ganesan Vadamalai ◽  
Ghulam Kadir Ahmad Parveez ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Insect Pests ◽  
Plant Protection ◽  
Pests And Diseases ◽  
The Future
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