Abstract
The roles of mitogen-activated protein kinases (MAPKs) in plant-fungal pathogenic interactions are poorly understood in crops. Here, microscopic, phenotypic, proteomic, and biochemical analyses revealed that roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection. When co-cultured with roots of the HvMPK3 KO lines, F. graminearum hyphae were excluded to the extracellular space, the growth pattern of extracellular hyphae was considerably deregulated, mycelia development was less efficient, and number of appressoria-like structures and their penetration potential were substantially reduced. Intracellular penetration of hyphae was preceded by the massive production of reactive oxygen species (ROS) in attacked cells of the wild-type (WT), but ROS production was mitigated in the HvMPK3 KO lines. Suppression of ROS production in these lines coincided with elevated abundance of catalase (CAT) and ascorbate peroxidase (APX). Moreover, differential proteomic analysis revealed downregulation of several defense-related proteins in WT, and the upregulation of pathogenesis-related protein 1 (PR-1) and cysteine proteases in HvMPK3 KO lines. Proteins involved in suberin formation, such as peroxidases, lipid transfer proteins (LTPs), and the GDSL esterase/lipase (containing "GDSL" aminosequence motif) were differentially regulated in HvMPK3 KO lines after F. graminearum inoculation. Consistent with proteomic analysis, microscopic observations showed enhanced suberin accumulation in roots of HvMPK3 KO lines, most likely contributing to the arrested infection by F. graminearum. These results suggest that TALEN-based knockout of HvMPK3 leads to barley root resistance against Fusarium root rot.
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Claims 13
Claim · model-written summary
TALEN-based knockout of HvMPK3 gives barley roots resistance against Fusarium graminearum infection.
- Taxon
- barley
.jpg?width=60)
Hordeum vulgaree0 - Material
- HvMPK3 KOe0
- Organ / stage
- rootse0
- Treatment
- (TALEN)-based knockoute0
- Interactor
- Fusarium graminearum
_on_Corn_(Zea_mays)_at_Waimea%2C_Hawaii.jpg?width=60)
Fusarium graminearume0pathogen - Gene
- HvMPK3e0
- Outcome
- resistant against Fusarium graminearum infectione0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
.jpg?width=500)
_on_Corn_(Zea_mays)_at_Waimea%2C_Hawaii.jpg?width=500)
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
Claim · model-written summary
Fusarium graminearum hyphae were excluded to the extracellular space when co-cultured with HvMPK3 KO barley roots.
- Taxon
- barleyHordeum vulgaree0
- Treatment
- co-cultured with roots of the HvMPK3 KO linese1
- Interactor
- F. graminearumFusarium graminearume1pathogen
- Outcome
- F. graminearum hyphae were excluded to the extracellular spacee1
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
e1“When co-cultured with roots of the HvMPK3 KO lines, F. graminearum hyphae were excluded to the extracellular space, the growth pattern of extracellular hyphae was considerably deregulated, mycelia development was less efficient, and number of appressoria-like structures and their penetration potential were substantially reduced”
Claim · model-written summary
In HvMPK3 KO barley roots co-cultured with F. graminearum, the growth pattern of extracellular hyphae was considerably deregulated.
- Taxon
- barleyHordeum vulgaree0
- Treatment
- co-cultured with roots of the HvMPK3 KO linese1
- Interactor
- F. graminearumFusarium graminearume1pathogen
- Outcome
- growth pattern of extracellular hyphae was considerably deregulatede1
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
e1“When co-cultured with roots of the HvMPK3 KO lines, F. graminearum hyphae were excluded to the extracellular space, the growth pattern of extracellular hyphae was considerably deregulated, mycelia development was less efficient, and number of appressoria-like structures and their penetration potential were substantially reduced”
Claim · model-written summary
Mycelia development of F. graminearum was less efficient on HvMPK3 KO barley roots.
- Taxon
- barleyHordeum vulgaree0
- Treatment
- co-cultured with roots of the HvMPK3 KO linese1
- Interactor
- F. graminearumFusarium graminearume1pathogen
- Outcome
- mycelia developmente1
- Direction
- less efficiente1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
e1“When co-cultured with roots of the HvMPK3 KO lines, F. graminearum hyphae were excluded to the extracellular space, the growth pattern of extracellular hyphae was considerably deregulated, mycelia development was less efficient, and number of appressoria-like structures and their penetration potential were substantially reduced”
Claim · model-written summary
Appressoria-like structures of F. graminearum and their penetration potential were substantially reduced on HvMPK3 KO barley roots.
- Taxon
- barleyHordeum vulgaree0
- Treatment
- co-cultured with roots of the HvMPK3 KO linese1
- Interactor
- F. graminearumFusarium graminearume1pathogen
- Outcome
- number of appressoria-like structures and their penetration potentiale1
- Direction
- substantially reducede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
e1“When co-cultured with roots of the HvMPK3 KO lines, F. graminearum hyphae were excluded to the extracellular space, the growth pattern of extracellular hyphae was considerably deregulated, mycelia development was less efficient, and number of appressoria-like structures and their penetration potential were substantially reduced”
Claim · model-written summary
In wild-type barley, intracellular penetration by F. graminearum hyphae was preceded by a massive production of ROS in attacked cells.
- Taxon
- barleyHordeum vulgaree0
- Material
- wild-type (WT)e5
- Outcome
- massive production of reactive oxygen species (ROS) in attacked cellse5
- Qualifier
- Intracellular penetration of hyphae was preceded bye5
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
e5“Intracellular penetration of hyphae was preceded by the massive production of reactive oxygen species (ROS) in attacked cells of the wild-type (WT), but ROS production was mitigated in the HvMPK3 KO lines”
Claim · model-written summary
ROS production was mitigated in HvMPK3 KO barley lines relative to wild-type.
- Taxon
- barleyHordeum vulgaree0
- Material
- HvMPK3 KO linese5
- Treatment
- (TALEN)-based knockoute0
- Outcome
- ROS productione5
- Direction
- mitigatede5
- Comparator
- wild-type (WT)e5
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
e5“Intracellular penetration of hyphae was preceded by the massive production of reactive oxygen species (ROS) in attacked cells of the wild-type (WT), but ROS production was mitigated in the HvMPK3 KO lines”
Claim · model-written summary
Suppression of ROS production in HvMPK3 KO barley coincided with elevated abundance of catalase (CAT) and ascorbate peroxidase (APX).
- Taxon
- barleyHordeum vulgaree0
- Treatment
- Suppression of ROS productione6
- Gene
- catalase (CAT) and ascorbate peroxidase (APX)e6
- Outcome
- abundance of catalase (CAT) and ascorbate peroxidase (APX)e6
- Direction
- elevatede6
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
e6“Suppression of ROS production in these lines coincided with elevated abundance of catalase (CAT) and ascorbate peroxidase (APX)”
Claim · model-written summary
Several defense-related proteins were downregulated in wild-type barley roots.
- Taxon
- barleyHordeum vulgaree0
- Material
- WTe7
- Outcome
- defense-related proteinse7
- Direction
- downregulatione7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
e7“differential proteomic analysis revealed downregulation of several defense-related proteins in WT, and the upregulation of pathogenesis-related protein 1 (PR-1) and cysteine proteases in HvMPK3 KO lines”
Claim · model-written summary
Pathogenesis-related protein 1 (PR-1) and cysteine proteases were upregulated in HvMPK3 KO barley relative to wild-type.
- Taxon
- barleyHordeum vulgaree0
- Treatment
- (TALEN)-based knockoute0
- Gene
- pathogenesis-related protein 1 (PR-1)e7
- Outcome
- pathogenesis-related protein 1 (PR-1) and cysteine proteasese7
- Direction
- upregulatione7
- Comparator
- WTe7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
e7“differential proteomic analysis revealed downregulation of several defense-related proteins in WT, and the upregulation of pathogenesis-related protein 1 (PR-1) and cysteine proteases in HvMPK3 KO lines”
Claim · model-written summary
Proteins involved in suberin formation were differentially regulated in HvMPK3 KO barley lines after F. graminearum inoculation.
- Taxon
- barleyHordeum vulgaree0
- Material
- HvMPK3 KO linese2
- Treatment
- F. graminearum inoculatione2
- Interactor
- F. graminearumFusarium graminearume2pathogen
- Gene
- peroxidases, lipid transfer proteins (LTPs), and the GDSL esterase/lipasee2
- Outcome
- proteins involved in suberin formatione2
- Direction
- differentially regulatede2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
e2“Proteins involved in suberin formation, such as peroxidases, lipid transfer proteins (LTPs), and the GDSL esterase/lipase (containing "GDSL" aminosequence motif) were differentially regulated in HvMPK3 KO lines after F. graminearum inoculation”
Claim · model-written summary
Suberin accumulation was enhanced in roots of HvMPK3 KO barley.
- Taxon
- barleyHordeum vulgaree0
- Material
- HvMPK3 KO linese3
- Organ / stage
- rootse3
- Treatment
- (TALEN)-based knockoute0
- Outcome
- suberin accumulatione3
- Direction
- enhancede3
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
e3“microscopic observations showed enhanced suberin accumulation in roots of HvMPK3 KO lines, most likely contributing to the arrested infection by F. graminearum”
Claim · model-written summary
Enhanced suberin accumulation in roots of HvMPK3 KO barley most likely contributes to arrested F. graminearum infection.
- Taxon
- barleyHordeum vulgaree0
- Interactor
- F. graminearumFusarium graminearume3pathogen
- Outcome
- arrested infection by F. graminearume3
- Qualifier
- enhanced suberin accumulation in roots of HvMPK3 KO linese3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“roots of independent transcription activator-like effector nuclease (TALEN)-based knockout lines of barley (Hordeum vulgare L.) MAPK 3 (HvMPK3 KO) were resistant against Fusarium graminearum infection”
e3“microscopic observations showed enhanced suberin accumulation in roots of HvMPK3 KO lines, most likely contributing to the arrested infection by F. graminearum”
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