Host-Induced Gene Silencing of a Sclerotinia sclerotiorum oxaloacetate acetylhydrolase Using Bean Pod Mottle Virus as a Vehicle Reduces Disease on Soybean

A lack of complete resistance in the current germplasm complicates the management of Sclerotinia stem rot (SSR) caused by Sclerotinia sclerotiorum in soybean. In this study, we used bean pod mottle virus (BPMV) as a vehicle to down-regulate expression of a key enzyme in the production of an importan...

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Main Authors: Megan McCaghey, Dandan Shao, Jake Kurcezewski, Ally Lindstrom, Ashish Ranjan, Steven A. Whitham, Shawn P. Conley, Brett Williams, Damon L. Smith, Mehdi Kabbage
Format: Article
Language:English
Published: Frontiers Media S.A. 2021-07-01
Series:Frontiers in Plant Science
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Online Access:https://www.frontiersin.org/articles/10.3389/fpls.2021.677631/full
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author Megan McCaghey
Dandan Shao
Jake Kurcezewski
Ally Lindstrom
Ashish Ranjan
Steven A. Whitham
Shawn P. Conley
Brett Williams
Damon L. Smith
Mehdi Kabbage
author_facet Megan McCaghey
Dandan Shao
Jake Kurcezewski
Ally Lindstrom
Ashish Ranjan
Steven A. Whitham
Shawn P. Conley
Brett Williams
Damon L. Smith
Mehdi Kabbage
author_sort Megan McCaghey
collection DOAJ
description A lack of complete resistance in the current germplasm complicates the management of Sclerotinia stem rot (SSR) caused by Sclerotinia sclerotiorum in soybean. In this study, we used bean pod mottle virus (BPMV) as a vehicle to down-regulate expression of a key enzyme in the production of an important virulence factor in S. sclerotiorum, oxalic acid (OA). Specifically, we targeted a gene encoding oxaloacetate acetylhydrolase (Ssoah1), because Ssoah1 deletion mutants are OA deficient and non-pathogenic on soybean. We first established that S. sclerotiorum can uptake environmental RNAs by monitoring the translocation of Cy3-labeled double-stranded and small interfering RNA (ds/siRNAs) into fungal hyphae using fluorescent confocal microscopy. This translocation led to a significant decrease in Ssoah1 transcript levels in vitro. Inoculation of soybean plants with BPMV vectors targeting Ssoah1 (pBPMV-OA) also led to decreased expression of Ssoah1. Importantly, pBPMV-OA inoculated plants showed enhanced resistance to S. sclerotiorum compared to empty-vector control plants. Our combined results provide evidence supporting the use of HIGS and exogenous applications of ds/siRNAs targeting virulence factors such as OA as viable strategies for the control of SSR in soybean and as discovery tools that can be used to identify previously unknown virulence factors.
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spelling doaj.art-feda19f94e1046879c0b9f62662888542022-12-21T18:22:22ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2021-07-011210.3389/fpls.2021.677631677631Host-Induced Gene Silencing of a Sclerotinia sclerotiorum oxaloacetate acetylhydrolase Using Bean Pod Mottle Virus as a Vehicle Reduces Disease on SoybeanMegan McCaghey0Dandan Shao1Jake Kurcezewski2Ally Lindstrom3Ashish Ranjan4Steven A. Whitham5Shawn P. Conley6Brett Williams7Damon L. Smith8Mehdi Kabbage9Department of Plant Pathology, University of Wisconsin-Madison, Madison, WI, United StatesDepartment of Plant Pathology, University of Wisconsin-Madison, Madison, WI, United StatesDepartment of Plant Pathology, University of Wisconsin-Madison, Madison, WI, United StatesDepartment of Plant Pathology, University of Wisconsin-Madison, Madison, WI, United StatesDepartment of Plant Pathology, University of Minnesota, St. Paul, MN, United StatesDepartment of Plant Pathology and Microbiology, Iowa State University, Ames, IA, United StatesDepartment of Agronomy, University of Wisconsin-Madison, Madison, WI, United StatesCentre for Tropical Crops and Biocommodities, Queensland University of Technology, Brisbane, QLD, AustraliaDepartment of Plant Pathology, University of Wisconsin-Madison, Madison, WI, United StatesDepartment of Plant Pathology, University of Wisconsin-Madison, Madison, WI, United StatesA lack of complete resistance in the current germplasm complicates the management of Sclerotinia stem rot (SSR) caused by Sclerotinia sclerotiorum in soybean. In this study, we used bean pod mottle virus (BPMV) as a vehicle to down-regulate expression of a key enzyme in the production of an important virulence factor in S. sclerotiorum, oxalic acid (OA). Specifically, we targeted a gene encoding oxaloacetate acetylhydrolase (Ssoah1), because Ssoah1 deletion mutants are OA deficient and non-pathogenic on soybean. We first established that S. sclerotiorum can uptake environmental RNAs by monitoring the translocation of Cy3-labeled double-stranded and small interfering RNA (ds/siRNAs) into fungal hyphae using fluorescent confocal microscopy. This translocation led to a significant decrease in Ssoah1 transcript levels in vitro. Inoculation of soybean plants with BPMV vectors targeting Ssoah1 (pBPMV-OA) also led to decreased expression of Ssoah1. Importantly, pBPMV-OA inoculated plants showed enhanced resistance to S. sclerotiorum compared to empty-vector control plants. Our combined results provide evidence supporting the use of HIGS and exogenous applications of ds/siRNAs targeting virulence factors such as OA as viable strategies for the control of SSR in soybean and as discovery tools that can be used to identify previously unknown virulence factors.https://www.frontiersin.org/articles/10.3389/fpls.2021.677631/fullhost induced gene silencingRNAiSclerotinia sclerotiorumsoybeanoxalic acidSsoah1
spellingShingle Megan McCaghey
Dandan Shao
Jake Kurcezewski
Ally Lindstrom
Ashish Ranjan
Steven A. Whitham
Shawn P. Conley
Brett Williams
Damon L. Smith
Mehdi Kabbage
Host-Induced Gene Silencing of a Sclerotinia sclerotiorum oxaloacetate acetylhydrolase Using Bean Pod Mottle Virus as a Vehicle Reduces Disease on Soybean
Frontiers in Plant Science
host induced gene silencing
RNAi
Sclerotinia sclerotiorum
soybean
oxalic acid
Ssoah1
title Host-Induced Gene Silencing of a Sclerotinia sclerotiorum oxaloacetate acetylhydrolase Using Bean Pod Mottle Virus as a Vehicle Reduces Disease on Soybean
title_full Host-Induced Gene Silencing of a Sclerotinia sclerotiorum oxaloacetate acetylhydrolase Using Bean Pod Mottle Virus as a Vehicle Reduces Disease on Soybean
title_fullStr Host-Induced Gene Silencing of a Sclerotinia sclerotiorum oxaloacetate acetylhydrolase Using Bean Pod Mottle Virus as a Vehicle Reduces Disease on Soybean
title_full_unstemmed Host-Induced Gene Silencing of a Sclerotinia sclerotiorum oxaloacetate acetylhydrolase Using Bean Pod Mottle Virus as a Vehicle Reduces Disease on Soybean
title_short Host-Induced Gene Silencing of a Sclerotinia sclerotiorum oxaloacetate acetylhydrolase Using Bean Pod Mottle Virus as a Vehicle Reduces Disease on Soybean
title_sort host induced gene silencing of a sclerotinia sclerotiorum oxaloacetate acetylhydrolase using bean pod mottle virus as a vehicle reduces disease on soybean
topic host induced gene silencing
RNAi
Sclerotinia sclerotiorum
soybean
oxalic acid
Ssoah1
url https://www.frontiersin.org/articles/10.3389/fpls.2021.677631/full
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