Related Experiment Video
Updated: Aug 2, 2025
Lignin Down-regulation of Zea mays via dsRNAi and Klason Lignin Analysis
Published on: July 23, 2014
Control of white mold (Sclerotinia sclerotiorum) through plant-mediated RNA interference
Philip L Walker1, Dylan J Ziegler1, Shayna Giesbrecht1
1Department of Biological Sciences, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada.
Abstract:
The causative agent of white mold, Sclerotinia sclerotiorum, is capable of infecting over 600 plant species and is responsible for significant crop losses across the globe. Control is currently dependent on broad-spectrum chemical agents that can negatively impact the agroecological environment, presenting a need to develop alternative control measures. In this study, we developed transgenic Arabidopsis thaliana (AT1703) expressing hairpin (hp)RNA to silence S. sclerotiorum ABHYDROLASE-3 and slow infection through host induced gene silencing (HIGS). Leaf infection assays show reduced S. sclerotiorum lesion size, fungal load, and ABHYDROLASE-3 transcript abundance in AT1703 compared to wild-type Col-0. To better understand how HIGS influences host-pathogen interactions, we performed global RNA sequencing on AT1703 and wild-type Col-0 directly at the site of S. sclerotiorum infection. RNA sequencing data reveals enrichment of the salicylic acid (SA)-mediated systemic acquired resistance (SAR) pathway, as well as transcription factors predicted to regulate plant immunity. Using RT-qPCR, we identified predicted interacting partners of ABHYDROLASE-3 in the polyamine synthesis pathway of S. sclerotiorum that demonstrate co-reduction with ABHYDROLASE-3 transcript levels during infection. Together, these results demonstrate the utility of HIGS technology in slowing S. sclerotiorum infection and provide insight into the role of ABHYDROLASE-3 in the A. thaliana-S. sclerotiorum pathosystem.
Insights
Host-induced gene silencing (HIGS) effectively reduced white mold pathogen Sclerotinia sclerotiorum infection in Arabidopsis. This novel approach targets the ABHYDROLASE-3 gene, offering a sustainable alternative to chemical controls.
Area of Science:
- Plant Pathology
- Molecular Biology
- Biotechnology
Background:
- White mold, caused by Sclerotinia sclerotiorum, devastates over 600 plant species, leading to significant global crop losses.
- Current control relies on broad-spectrum chemicals, posing risks to the agroecological environment.
- Development of sustainable, targeted control measures is crucial.
Purpose of the Study:
- To develop and evaluate host-induced gene silencing (HIGS) as a strategy to control Sclerotinia sclerotiorum infection.
- To investigate the role of the S. sclerotiorum ABHYDROLASE-3 gene in the pathosystem.
- To elucidate the molecular mechanisms underlying HIGS-mediated resistance.
Main Methods:
- Generated transgenic Arabidopsis thaliana (AT1703) expressing hairpin RNA to silence S. sclerotiorum ABHYDROLASE-3 via HIGS.
- Performed leaf infection assays to quantify disease progression (lesion size, fungal load).
- Utilized RNA sequencing and RT-qPCR to analyze host and pathogen gene expression changes during infection.
Main Results:
- Transgenic AT1703 plants exhibited significantly reduced S. sclerotiorum lesion size and fungal load compared to wild-type.
- ABHYDROLASE-3 transcript levels were reduced in infected AT1703 plants.
- RNA sequencing revealed activation of salicylic acid-mediated systemic acquired resistance (SAR) and immune-related transcription factors.
- Identified S. sclerotiorum polyamine synthesis pathway genes co-reduced with ABHYDROLASE-3.
Conclusions:
- HIGS is a viable technology for slowing Sclerotinia sclerotiorum infection in plants.
- ABHYDROLASE-3 plays a role in the Arabidopsis-S. sclerotiorum pathosystem.
- HIGS modulates plant immunity pathways and impacts pathogen gene expression, offering a novel control strategy.
Related Concept Videos
02:15Experimental RNAi
01:23RNA Interference
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
02:30siRNA - Small Interfering RNAs
In the cytoplasm, siRNA is processed from a double-stranded RNA, which comes from either endogenous DNA transcription or exogenous sources like a virus. This double-stranded RNA is then cleaved by the...