Abstract
We successfully implemented virus-induced gene silencing (VIGS) in barley (Hordeum vulgare) for the functional characterization of genes required for Mla13-mediated resistance toward the biotrophic barley pathogen Blumeria graminis f. sp. hordei. Initially, barley cultivars were screened for their ability to host the barley stripe mosaic virus (BSMV)-VIGS vector by allowing its replication and systemic movement without causing excessive symptoms. Phytoene desaturase silencing leading to photobleaching was used as a phenotypic marker alongside reverse transcription-PCR data to characterize the silencing response at the molecular level. Barley cultivar Clansman, harboring the Mla13 resistance gene, was chosen as the most suitable host for BSMV-VIGS-based functional characterization of Rar1, Sgt1, and Hsp90 in the Mla-mediated resistance toward powdery mildew. BSMV-induced gene silencing of these candidate genes, which are associated in many but not all race-specific pathways, proved to be robust and could be detected at both mRNA and protein levels for up to 21 d postinoculation. Systemic silencing was observed not only in the newly developed leaves from the main stem but also in axillary shoots. By examining fungal development from an incompatible mildew strain carrying the cognate Avr13 gene on plants BSMV silenced for Rar1, Sgt1, and Hsp90, a resistance-breaking phenotype was observed, while plants infected with BSMV control constructs remained resistant. We demonstrate that Hsp90 is a required component for Mla13-mediated race-specific resistance and that BSMV-induced VIGS is a powerful tool to characterize genes involved in pathogen resistance in barley.
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Claims 13
Claim · model-written summary
VIGS was implemented in barley to characterize genes for Mla13 resistance.
- Taxon
- barley
.jpg?width=60)
Hordeum vulgaree0 - Interactor
- Blumeria graminis f. sp. hordeiBlumeria hordeie0pathogen
- Outcome
- functional characterization of genes required for Mla13-mediated resistancee0
- Value
- Mla13e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
.jpg?width=500)
Verbatim
e0“We successfully implemented virus-induced gene silencing (VIGS) in barley (Hordeum vulgare) for the functional characterization of genes required for Mla13-mediated resistance toward the biotrophic barley pathogen Blumeria graminis f. sp. hordei.”
Claim · model-written summary
Phytoene desaturase silencing produced photobleaching as a marker.
- Taxon
- BarleyHordeum vulgaret0
- Gene
- Phytoene desaturasee0
- Outcome
- photobleachinge0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Phytoene desaturase silencing leading to photobleaching was used as a phenotypic marker alongside reverse transcription-PCR data to characterize the silencing response at the molecular level.”
t0“Barley”
Claim · model-written summary
Clansman was selected as the most suitable BSMV-VIGS host.
- Taxon
- BarleyHordeum vulgaree0
- Material
- Clansmane0
- Gene
- Mla13e0
- Outcome
- most suitable hoste0
- Comparator
- BSMV-VIGS-based functional characterizatione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Barley cultivar Clansman, harboring the Mla13 resistance gene, was chosen as the most suitable host for BSMV-VIGS-based functional characterization of Rar1, Sgt1, and Hsp90 in the Mla-mediated resistance toward powdery mildew.”
Claim · model-written summary
BSMV silenced Rar1 robustly at mRNA and protein levels for up to 21 days.
- Taxon
- BarleyHordeum vulgaret0
- Gene
- Rar1e1
- Outcome
- gene silencinge0
- Value
- 21 d postinoculatione0
- Qualifier
- both mRNA and protein levelse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“BSMV-induced gene silencing of these candidate genes, which are associated in many but not all race-specific pathways, proved to be robust and could be detected at both mRNA and protein levels for up to 21 d postinoculation.”
e1“characterization of Rar1, Sgt1, and Hsp90”
t0“Barley”
Claim · model-written summary
BSMV silenced Sgt1 robustly at mRNA and protein levels for up to 21 days.
- Taxon
- BarleyHordeum vulgaret0
- Gene
- Sgt1e1
- Outcome
- gene silencinge0
- Value
- 21 d postinoculatione0
- Qualifier
- both mRNA and protein levelse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“BSMV-induced gene silencing of these candidate genes, which are associated in many but not all race-specific pathways, proved to be robust and could be detected at both mRNA and protein levels for up to 21 d postinoculation.”
e1“characterization of Rar1, Sgt1, and Hsp90”
t0“Barley”
Claim · model-written summary
BSMV silenced Hsp90 robustly at mRNA and protein levels for up to 21 days.
- Taxon
- BarleyHordeum vulgaret0
- Gene
- Hsp90e1
- Outcome
- gene silencinge0
- Value
- 21 d postinoculatione0
- Qualifier
- both mRNA and protein levelse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“BSMV-induced gene silencing of these candidate genes, which are associated in many but not all race-specific pathways, proved to be robust and could be detected at both mRNA and protein levels for up to 21 d postinoculation.”
e1“characterization of Rar1, Sgt1, and Hsp90”
t0“Barley”
Claim · model-written summary
Systemic silencing occurred in newly developed main-stem leaves.
- Taxon
- BarleyHordeum vulgaret0
- Organ / stage
- newly developed leaves from the main steme0
- Outcome
- Systemic silencinge0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Systemic silencing was observed not only in the newly developed leaves from the main stem but also in axillary shoots.”
t0“Barley”
Claim · model-written summary
Systemic silencing occurred in axillary shoots.
- Taxon
- BarleyHordeum vulgaret0
- Organ / stage
- axillary shootse0
- Outcome
- Systemic silencinge0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Systemic silencing was observed not only in the newly developed leaves from the main stem but also in axillary shoots.”
t0“Barley”
Claim · model-written summary
Silencing Rar1 caused a resistance-breaking phenotype against incompatible mildew.
- Taxon
- BarleyHordeum vulgaret0
- Treatment
- BSMV silenced for Rar1e0
- Interactor
- incompatible mildew straine0pathogen
- Gene
- Rar1e0
- Outcome
- resistance-breaking phenotypee0
- Comparator
- BSMV control constructse0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“By examining fungal development from an incompatible mildew strain carrying the cognate Avr13 gene on plants BSMV silenced for Rar1, Sgt1, and Hsp90, a resistance-breaking phenotype was observed, while plants infected with BSMV control constructs remained resistant.”
t0“Barley”
Claim · model-written summary
Silencing Sgt1 caused a resistance-breaking phenotype against incompatible mildew.
- Taxon
- BarleyHordeum vulgaret0
- Treatment
- BSMV silenced for Rar1, Sgt1, and Hsp90e0
- Interactor
- incompatible mildew straine0pathogen
- Gene
- Sgt1e0
- Outcome
- resistance-breaking phenotypee0
- Comparator
- BSMV control constructse0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“By examining fungal development from an incompatible mildew strain carrying the cognate Avr13 gene on plants BSMV silenced for Rar1, Sgt1, and Hsp90, a resistance-breaking phenotype was observed, while plants infected with BSMV control constructs remained resistant.”
t0“Barley”
Claim · model-written summary
Silencing Hsp90 caused a resistance-breaking phenotype against incompatible mildew.
- Taxon
- BarleyHordeum vulgaret0
- Treatment
- BSMV silenced for Rar1, Sgt1, and Hsp90e0
- Interactor
- incompatible mildew straine0pathogen
- Gene
- Hsp90e0
- Outcome
- resistance-breaking phenotypee0
- Comparator
- BSMV control constructse0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“By examining fungal development from an incompatible mildew strain carrying the cognate Avr13 gene on plants BSMV silenced for Rar1, Sgt1, and Hsp90, a resistance-breaking phenotype was observed, while plants infected with BSMV control constructs remained resistant.”
t0“Barley”
Claim · model-written summary
Hsp90 is required for Mla13-mediated race-specific resistance.
- Taxon
- barleyHordeum vulgaree0
- Gene
- Hsp90e0
- Outcome
- Mla13-mediated race-specific resistancee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“We demonstrate that Hsp90 is a required component for Mla13-mediated race-specific resistance and that BSMV-induced VIGS is a powerful tool to characterize genes involved in pathogen resistance in barley.”
Claim · model-written summary
BSMV-induced VIGS is a powerful tool for characterizing barley resistance genes.
- Taxon
- barleyHordeum vulgaree0
- Outcome
- powerful tool to characterize genes involved in pathogen resistancee0
- Value
- BSMV-induced VIGSe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“We demonstrate that Hsp90 is a required component for Mla13-mediated race-specific resistance and that BSMV-induced VIGS is a powerful tool to characterize genes involved in pathogen resistance in barley.”
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loss of function · disease · Hordeum vulgare (barley)Positive and negative results
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