Abstract
Abstract The mitogen-activated protein kinase kinase kinase kinase (M4K) family is evolutionarily conserved across plants and animals. In Arabidopsis, the protein kinase SIK1, an M4K member, is known to positively modulate reactive oxygen species (ROS) production during pattern-triggered immunity (PTI) by stabilizing BIK1, a key receptor-like cytoplasmic kinase (RLCK). While homologs of SIK1 exhibit conserved protein domain architectures across a range of land plants, their functional conservation remains incompletely understood. This study investigates the functional conservation and divergence of SIK1 homologs, focusing particularly on NbM4K3 in Nicotiana benthamiana . Silencing NbM4K3 resulted in an impairment of the flg22-induced ROS burst and expression of PTI marker genes. Additionally, silencing NbM4K3 led to diminished protein accumulation of RLCKs, while overexpression of the RLCKs prominently enhanced the flg22-induced ROS burst in NbM4K3 -silenced plants. Furthermore, NbM4K3 -silenced plants exhibited a compromised hypersensitive response (HR), reduced ROS accumulation, and diminished expression of effector-triggered immunity (ETI) marker genes when challenged with the avirulent strains Ralstonia solanacearum GMI1000 and Pseudomonas syringae DC3000, suggesting that NbM4K3 is a positive regulator of ETI. The attenuated HR phenotype observed in NbM4K3 -silenced plants upon expression of RipP1 or RipE1, two avirulent type III effectors of GMI1000, further supports the affirmative role of NbM4K3 in ETI. In summary, our data indicate that the M4K NbM4K3 positively regulates both PTI and ETI in N. benthamiana , potentially by stabilizing RLCKs. These findings not only strengthen the role of M4K family in plant immunity but also suggest its potential in improving disease resistance in plants.
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Claims 10
Claim · model-written summary
NbM4K3 silencing impaired the flg22-induced ROS burst.
- Taxon
- Nicotiana benthamiana
Nicotiana benthamianae1 - Treatment
- Silencing NbM4K3e0
- Gene
- NbM4K3e0
- Outcome
- flg22-induced ROS burste0
- Direction
- impairmente0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Silencing NbM4K3 resulted in an impairment of the flg22-induced ROS burst and expression of PTI marker genes.”
e1“Nicotiana benthamiana”
Claim · model-written summary
NbM4K3 silencing impaired PTI marker-gene expression.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- Silencing NbM4K3e0
- Gene
- NbM4K3e0
- Outcome
- expression of PTI marker genese0
- Direction
- impairmente0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Silencing NbM4K3 resulted in an impairment of the flg22-induced ROS burst and expression of PTI marker genes.”
e1“Nicotiana benthamiana”
Claim · model-written summary
NbM4K3 silencing diminished RLCK protein accumulation.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- silencing NbM4K3e0
- Gene
- RLCKse0
- Outcome
- protein accumulation of RLCKse0
- Direction
- diminishede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“silencing NbM4K3 led to diminished protein accumulation of RLCKs”
e1“Nicotiana benthamiana”
Claim · model-written summary
RLCK overexpression enhanced the flg22 ROS burst in NbM4K3-silenced plants.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- overexpression of the RLCKse0
- Gene
- RLCKse0
- Outcome
- flg22-induced ROS burste0
- Direction
- enhancede0
- Qualifier
- NbM4K3 -silenced plantse0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“overexpression of the RLCKs prominently enhanced the flg22-induced ROS burst in NbM4K3 -silenced plants.”
e1“Nicotiana benthamiana”
Claim · model-written summary
NbM4K3 silencing compromised HR after challenge with Ralstonia solanacearum GMI1000.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- NbM4K3 -silencede0
- Interactor
- Ralstonia solanacearum GMI1000Ralstonia pseudosolanacearum GMI1000e0pathogen
- Outcome
- hypersensitive response (HR)e0
- Direction
- compromisede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“NbM4K3 -silenced plants exhibited a compromised hypersensitive response (HR), reduced ROS accumulation, and diminished expression of effector-triggered immunity (ETI) marker genes when challenged with the avirulent strains Ralstonia solanacearum GMI1000 and Pseudomonas syringae DC3000”
e1“Nicotiana benthamiana”
Claim · model-written summary
NbM4K3 silencing compromised HR after challenge with Pseudomonas syringae DC3000.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- NbM4K3 -silencede0
- Interactor
- Pseudomonas syringae DC3000Pseudomonas syringae pv. tomato str. DC3000e0pathogen
- Outcome
- hypersensitive response (HR)e0
- Direction
- compromisede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“NbM4K3 -silenced plants exhibited a compromised hypersensitive response (HR), reduced ROS accumulation, and diminished expression of effector-triggered immunity (ETI) marker genes when challenged with the avirulent strains Ralstonia solanacearum GMI1000 and Pseudomonas syringae DC3000”
e1“Nicotiana benthamiana”
Claim · model-written summary
NbM4K3 silencing reduced ROS accumulation after avirulent pathogen challenge.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- NbM4K3 -silencede0
- Interactor
- Ralstonia solanacearum GMI1000Ralstonia pseudosolanacearum GMI1000e0pathogen
- Second interactor
- Pseudomonas syringae DC3000Pseudomonas syringae pv. tomato str. DC3000e0pathogen
- Outcome
- ROS accumulatione0
- Direction
- reducede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“NbM4K3 -silenced plants exhibited a compromised hypersensitive response (HR), reduced ROS accumulation, and diminished expression of effector-triggered immunity (ETI) marker genes when challenged with the avirulent strains Ralstonia solanacearum GMI1000 and Pseudomonas syringae DC3000”
e1“Nicotiana benthamiana”
Claim · model-written summary
NbM4K3 silencing diminished ETI marker-gene expression after avirulent pathogen challenge.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- NbM4K3 -silencede0
- Interactor
- Ralstonia solanacearum GMI1000Ralstonia pseudosolanacearum GMI1000e0pathogen
- Second interactor
- Pseudomonas syringae DC3000Pseudomonas syringae pv. tomato str. DC3000e0pathogen
- Outcome
- expression of effector-triggered immunity (ETI) marker genese0
- Direction
- diminishede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“NbM4K3 -silenced plants exhibited a compromised hypersensitive response (HR), reduced ROS accumulation, and diminished expression of effector-triggered immunity (ETI) marker genes when challenged with the avirulent strains Ralstonia solanacearum GMI1000 and Pseudomonas syringae DC3000”
e1“Nicotiana benthamiana”
Claim · model-written summary
RipP1 expression attenuated HR in NbM4K3-silenced plants.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- expression of RipP1e0
- Gene
- NbM4K3e0
- Outcome
- HR phenotypee0
- Direction
- attenuatede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The attenuated HR phenotype observed in NbM4K3 -silenced plants upon expression of RipP1 or RipE1”
e1“Nicotiana benthamiana”
Claim · model-written summary
RipE1 expression attenuated HR in NbM4K3-silenced plants.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- RipE1e0
- Gene
- NbM4K3e0
- Outcome
- HR phenotypee0
- Direction
- attenuatede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The attenuated HR phenotype observed in NbM4K3 -silenced plants upon expression of RipP1 or RipE1”
e1“Nicotiana benthamiana”
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