Targeted Knockout of Eukaryotic Translation Initiation Factor 4E Confers Bymovirus Resistance in Winter Barley

The Eukaryotic Translation Initiation Factor 4E (EIF4E) is a well-known susceptibility factor for potyvirus infections in many plant species. The barley yellow mosaic virus disease, caused by the bymoviruses Barley yellow mosaic virus (BaYMV) and Barley mild mosaic virus (BaMMV), can lead to yield l...

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Autores principales: Robert Eric Hoffie, Ingrid Otto, Dragan Perovic, Nagaveni Budhagatapalli, Antje Habekuß, Frank Ordon, Jochen Kumlehn
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Publicado: Frontiers Media S.A. 2021
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Acceso en línea:https://doaj.org/article/b3e38729a6914286849256181a929a3c
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spelling oai:doaj.org-article:b3e38729a6914286849256181a929a3c2021-12-01T14:12:21ZTargeted Knockout of Eukaryotic Translation Initiation Factor 4E Confers Bymovirus Resistance in Winter Barley2673-343910.3389/fgeed.2021.784233https://doaj.org/article/b3e38729a6914286849256181a929a3c2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fgeed.2021.784233/fullhttps://doaj.org/toc/2673-3439The Eukaryotic Translation Initiation Factor 4E (EIF4E) is a well-known susceptibility factor for potyvirus infections in many plant species. The barley yellow mosaic virus disease, caused by the bymoviruses Barley yellow mosaic virus (BaYMV) and Barley mild mosaic virus (BaMMV), can lead to yield losses of up to 50% in winter barley. In autumn, the roots of young barley plants are infected by the soil-borne plasmodiophoraceous parasite Polymyxa graminis L. that serves as viral vector. Upon viral establishment and systemic spreading into the upper parts of the plants, yellow mosaics occur as first symptoms on leaves. In the further course of plant development, the disease entails leaf necrosis and increased susceptibility to frost damage. Thanks to the rym4 and rym5 allelic variants of the HvEIF4E gene, more than two thirds of current European winter barley cultivars are resistant to BaYMV and BaMMV. However, several strains of BaYMV and BaMMV have already overcome rym4- and rym5-mediated resistance. Accordingly, new resistance-conferring alleles are needed for barley breeding. Therefore, we performed targeted mutagenesis of the EIF4E gene by Cas9 endonuclease in BaMMV/BaYMV-susceptible winter barley cv. “Igri”. Small insertions were generated, resulting in a shift of the translational reading frame, thereby causing the loss-of-function of EIF4E. The mutations occurred in the homozygous state already in the primary mutants. Their progeny proved invariably homozygous and fully resistant to mechanical inoculation with BaMMV. EIF4E knockout plants showed normal growth habit and produced grains, yet exhibited a yield penalty.Robert Eric HoffieIngrid OttoDragan PerovicNagaveni BudhagatapalliAntje HabekußFrank OrdonJochen KumlehnFrontiers Media S.A.articleCas9CRISPRgenome editingRNA-guided endonucleasestargeted mutagenesisprotoplastsBiotechnologyTP248.13-248.65GeneticsQH426-470ENFrontiers in Genome Editing, Vol 3 (2021)
institution DOAJ
collection DOAJ
language EN
topic Cas9
CRISPR
genome editing
RNA-guided endonucleases
targeted mutagenesis
protoplasts
Biotechnology
TP248.13-248.65
Genetics
QH426-470
spellingShingle Cas9
CRISPR
genome editing
RNA-guided endonucleases
targeted mutagenesis
protoplasts
Biotechnology
TP248.13-248.65
Genetics
QH426-470
Robert Eric Hoffie
Ingrid Otto
Dragan Perovic
Nagaveni Budhagatapalli
Antje Habekuß
Frank Ordon
Jochen Kumlehn
Targeted Knockout of Eukaryotic Translation Initiation Factor 4E Confers Bymovirus Resistance in Winter Barley
description The Eukaryotic Translation Initiation Factor 4E (EIF4E) is a well-known susceptibility factor for potyvirus infections in many plant species. The barley yellow mosaic virus disease, caused by the bymoviruses Barley yellow mosaic virus (BaYMV) and Barley mild mosaic virus (BaMMV), can lead to yield losses of up to 50% in winter barley. In autumn, the roots of young barley plants are infected by the soil-borne plasmodiophoraceous parasite Polymyxa graminis L. that serves as viral vector. Upon viral establishment and systemic spreading into the upper parts of the plants, yellow mosaics occur as first symptoms on leaves. In the further course of plant development, the disease entails leaf necrosis and increased susceptibility to frost damage. Thanks to the rym4 and rym5 allelic variants of the HvEIF4E gene, more than two thirds of current European winter barley cultivars are resistant to BaYMV and BaMMV. However, several strains of BaYMV and BaMMV have already overcome rym4- and rym5-mediated resistance. Accordingly, new resistance-conferring alleles are needed for barley breeding. Therefore, we performed targeted mutagenesis of the EIF4E gene by Cas9 endonuclease in BaMMV/BaYMV-susceptible winter barley cv. “Igri”. Small insertions were generated, resulting in a shift of the translational reading frame, thereby causing the loss-of-function of EIF4E. The mutations occurred in the homozygous state already in the primary mutants. Their progeny proved invariably homozygous and fully resistant to mechanical inoculation with BaMMV. EIF4E knockout plants showed normal growth habit and produced grains, yet exhibited a yield penalty.
format article
author Robert Eric Hoffie
Ingrid Otto
Dragan Perovic
Nagaveni Budhagatapalli
Antje Habekuß
Frank Ordon
Jochen Kumlehn
author_facet Robert Eric Hoffie
Ingrid Otto
Dragan Perovic
Nagaveni Budhagatapalli
Antje Habekuß
Frank Ordon
Jochen Kumlehn
author_sort Robert Eric Hoffie
title Targeted Knockout of Eukaryotic Translation Initiation Factor 4E Confers Bymovirus Resistance in Winter Barley
title_short Targeted Knockout of Eukaryotic Translation Initiation Factor 4E Confers Bymovirus Resistance in Winter Barley
title_full Targeted Knockout of Eukaryotic Translation Initiation Factor 4E Confers Bymovirus Resistance in Winter Barley
title_fullStr Targeted Knockout of Eukaryotic Translation Initiation Factor 4E Confers Bymovirus Resistance in Winter Barley
title_full_unstemmed Targeted Knockout of Eukaryotic Translation Initiation Factor 4E Confers Bymovirus Resistance in Winter Barley
title_sort targeted knockout of eukaryotic translation initiation factor 4e confers bymovirus resistance in winter barley
publisher Frontiers Media S.A.
publishDate 2021
url https://doaj.org/article/b3e38729a6914286849256181a929a3c
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