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...
Main Authors: | , , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Frontiers Media S.A.
2021-07-01
|
Series: | Frontiers in Plant Science |
Subjects: | |
Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2021.677631/full |
_version_ | 1819152440442748928 |
---|---|
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. |
first_indexed | 2024-12-22T14:49:20Z |
format | Article |
id | doaj.art-feda19f94e1046879c0b9f6266288854 |
institution | Directory Open Access Journal |
issn | 1664-462X |
language | English |
last_indexed | 2024-12-22T14:49:20Z |
publishDate | 2021-07-01 |
publisher | Frontiers Media S.A. |
record_format | Article |
series | Frontiers in Plant Science |
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 |
work_keys_str_mv | AT meganmccaghey hostinducedgenesilencingofasclerotiniasclerotiorumoxaloacetateacetylhydrolaseusingbeanpodmottlevirusasavehiclereducesdiseaseonsoybean AT dandanshao hostinducedgenesilencingofasclerotiniasclerotiorumoxaloacetateacetylhydrolaseusingbeanpodmottlevirusasavehiclereducesdiseaseonsoybean AT jakekurcezewski hostinducedgenesilencingofasclerotiniasclerotiorumoxaloacetateacetylhydrolaseusingbeanpodmottlevirusasavehiclereducesdiseaseonsoybean AT allylindstrom hostinducedgenesilencingofasclerotiniasclerotiorumoxaloacetateacetylhydrolaseusingbeanpodmottlevirusasavehiclereducesdiseaseonsoybean AT ashishranjan hostinducedgenesilencingofasclerotiniasclerotiorumoxaloacetateacetylhydrolaseusingbeanpodmottlevirusasavehiclereducesdiseaseonsoybean AT stevenawhitham hostinducedgenesilencingofasclerotiniasclerotiorumoxaloacetateacetylhydrolaseusingbeanpodmottlevirusasavehiclereducesdiseaseonsoybean AT shawnpconley hostinducedgenesilencingofasclerotiniasclerotiorumoxaloacetateacetylhydrolaseusingbeanpodmottlevirusasavehiclereducesdiseaseonsoybean AT brettwilliams hostinducedgenesilencingofasclerotiniasclerotiorumoxaloacetateacetylhydrolaseusingbeanpodmottlevirusasavehiclereducesdiseaseonsoybean AT damonlsmith hostinducedgenesilencingofasclerotiniasclerotiorumoxaloacetateacetylhydrolaseusingbeanpodmottlevirusasavehiclereducesdiseaseonsoybean AT mehdikabbage hostinducedgenesilencingofasclerotiniasclerotiorumoxaloacetateacetylhydrolaseusingbeanpodmottlevirusasavehiclereducesdiseaseonsoybean |