Receptor-Like Kinases BAK1 and SOBIR1 Are Required for Necrotizing Activity of a Novel Group of Sclerotinia sclerotiorum Necrosis-Inducing Effectors

Sclerotinia sclerotiorum is a characteristic necrotrophic plant pathogen and is dependent on the induction of host cell death for nutrient acquisition. To identify necrosis-inducing effectors, the genome of S. sclerotiorum was scanned for genes encoding small, secreted, cysteine-rich proteins. These...

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Main Authors: Shirin Seifbarghi, Mohammad Hossein Borhan, Yangdou Wei, Lisong Ma, Cathy Coutu, Diana Bekkaoui, Dwayne D. Hegedus
Format: Article
Language:English
Published: Frontiers Media S.A. 2020-07-01
Series:Frontiers in Plant Science
Subjects:
Online Access:https://www.frontiersin.org/article/10.3389/fpls.2020.01021/full
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author Shirin Seifbarghi
Shirin Seifbarghi
Mohammad Hossein Borhan
Yangdou Wei
Lisong Ma
Cathy Coutu
Diana Bekkaoui
Dwayne D. Hegedus
Dwayne D. Hegedus
author_facet Shirin Seifbarghi
Shirin Seifbarghi
Mohammad Hossein Borhan
Yangdou Wei
Lisong Ma
Cathy Coutu
Diana Bekkaoui
Dwayne D. Hegedus
Dwayne D. Hegedus
author_sort Shirin Seifbarghi
collection DOAJ
description Sclerotinia sclerotiorum is a characteristic necrotrophic plant pathogen and is dependent on the induction of host cell death for nutrient acquisition. To identify necrosis-inducing effectors, the genome of S. sclerotiorum was scanned for genes encoding small, secreted, cysteine-rich proteins. These potential effectors were tested for their ability to induce necrosis in Nicotiana benthamiana via Agrobacterium-mediated expression and for cellular localization in host cells. Six novel proteins were discovered, of which all but one required a signal peptide for export to the apoplast for necrotizing activity. Virus-induced gene silencing revealed that the five necrosis-inducing effectors with a requirement for secretion also required the plant co-receptor-like kinases Brassinosteroid Insensitive 1-Associated Receptor Kinase 1 (BAK1) and Suppressor of BAK1-Interacting Receptor-like Kinase 1 (SOBIR1) for the induction of necrosis. S. sclerotiorum necrosis-inducing effector 2 (SsNE2) represented a new class of necrosis-inducing proteins as orthologs were identified in several other phytopathogenic fungi that were also capable of inducing necrosis. Substitution of conserved cysteine residues with alanine reduced, but did not abolish, the necrotizing activity of SsNE2 and full-length protein was required for function as peptides spanning the entire protein were unable to induce necrosis. These results illustrate the importance of necrosis-inducing effectors for S. sclerotiorum virulence and the role of host extracellular receptor(s) in effector-triggered susceptibility to this pathogen.
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spelling doaj.art-3adecee47fbe4c08825ce365f8e038822022-12-22T00:59:47ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2020-07-011110.3389/fpls.2020.01021527257Receptor-Like Kinases BAK1 and SOBIR1 Are Required for Necrotizing Activity of a Novel Group of Sclerotinia sclerotiorum Necrosis-Inducing EffectorsShirin Seifbarghi0Shirin Seifbarghi1Mohammad Hossein Borhan2Yangdou Wei3Lisong Ma4Cathy Coutu5Diana Bekkaoui6Dwayne D. Hegedus7Dwayne D. Hegedus8Agriculture and Agri-Food Canada, Saskatoon, SK, CanadaDepartment of Biology, University of Saskatchewan, Saskatoon, SK, CanadaAgriculture and Agri-Food Canada, Saskatoon, SK, CanadaDepartment of Biology, University of Saskatchewan, Saskatoon, SK, CanadaAgriculture and Agri-Food Canada, Saskatoon, SK, CanadaAgriculture and Agri-Food Canada, Saskatoon, SK, CanadaAgriculture and Agri-Food Canada, Saskatoon, SK, CanadaAgriculture and Agri-Food Canada, Saskatoon, SK, CanadaDepartment of Food and Bioproduct Sciences, University of Saskatchewan, Saskatoon, SK, CanadaSclerotinia sclerotiorum is a characteristic necrotrophic plant pathogen and is dependent on the induction of host cell death for nutrient acquisition. To identify necrosis-inducing effectors, the genome of S. sclerotiorum was scanned for genes encoding small, secreted, cysteine-rich proteins. These potential effectors were tested for their ability to induce necrosis in Nicotiana benthamiana via Agrobacterium-mediated expression and for cellular localization in host cells. Six novel proteins were discovered, of which all but one required a signal peptide for export to the apoplast for necrotizing activity. Virus-induced gene silencing revealed that the five necrosis-inducing effectors with a requirement for secretion also required the plant co-receptor-like kinases Brassinosteroid Insensitive 1-Associated Receptor Kinase 1 (BAK1) and Suppressor of BAK1-Interacting Receptor-like Kinase 1 (SOBIR1) for the induction of necrosis. S. sclerotiorum necrosis-inducing effector 2 (SsNE2) represented a new class of necrosis-inducing proteins as orthologs were identified in several other phytopathogenic fungi that were also capable of inducing necrosis. Substitution of conserved cysteine residues with alanine reduced, but did not abolish, the necrotizing activity of SsNE2 and full-length protein was required for function as peptides spanning the entire protein were unable to induce necrosis. These results illustrate the importance of necrosis-inducing effectors for S. sclerotiorum virulence and the role of host extracellular receptor(s) in effector-triggered susceptibility to this pathogen.https://www.frontiersin.org/article/10.3389/fpls.2020.01021/fullSclerotinia sclerotiorumcell deathnecrosisreceptorsvirus-induced gene silencing
spellingShingle Shirin Seifbarghi
Shirin Seifbarghi
Mohammad Hossein Borhan
Yangdou Wei
Lisong Ma
Cathy Coutu
Diana Bekkaoui
Dwayne D. Hegedus
Dwayne D. Hegedus
Receptor-Like Kinases BAK1 and SOBIR1 Are Required for Necrotizing Activity of a Novel Group of Sclerotinia sclerotiorum Necrosis-Inducing Effectors
Frontiers in Plant Science
Sclerotinia sclerotiorum
cell death
necrosis
receptors
virus-induced gene silencing
title Receptor-Like Kinases BAK1 and SOBIR1 Are Required for Necrotizing Activity of a Novel Group of Sclerotinia sclerotiorum Necrosis-Inducing Effectors
title_full Receptor-Like Kinases BAK1 and SOBIR1 Are Required for Necrotizing Activity of a Novel Group of Sclerotinia sclerotiorum Necrosis-Inducing Effectors
title_fullStr Receptor-Like Kinases BAK1 and SOBIR1 Are Required for Necrotizing Activity of a Novel Group of Sclerotinia sclerotiorum Necrosis-Inducing Effectors
title_full_unstemmed Receptor-Like Kinases BAK1 and SOBIR1 Are Required for Necrotizing Activity of a Novel Group of Sclerotinia sclerotiorum Necrosis-Inducing Effectors
title_short Receptor-Like Kinases BAK1 and SOBIR1 Are Required for Necrotizing Activity of a Novel Group of Sclerotinia sclerotiorum Necrosis-Inducing Effectors
title_sort receptor like kinases bak1 and sobir1 are required for necrotizing activity of a novel group of sclerotinia sclerotiorum necrosis inducing effectors
topic Sclerotinia sclerotiorum
cell death
necrosis
receptors
virus-induced gene silencing
url https://www.frontiersin.org/article/10.3389/fpls.2020.01021/full
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