Abstract
Our current understanding of pathogen-associated molecular pattern (PAMP)-triggered immunity signaling pathways in plants is limited due to the redundancy of several components or the lethality of mutants in Arabidopsis (Arabidopsis thaliana). To overcome this, we used a virus-induced gene silencing-based approach in combination with pharmacological studies to decipher links between early PAMP-triggered immunity events and their roles in immunity following PAMP perception in Nicotiana benthamiana. Two different calcium influx inhibitors suppressed the reactive oxygen species (ROS) burst: activation of the mitogen-activated protein kinases (MAPKs) and PAMP-induced gene expression. The calcium burst was unaffected in plants specifically silenced for components involved in ROS generation or for MAPKs activated by PAMP treatment. Importantly, the ROS burst still occurred in plants silenced for the two major defense-associated MAPK genes NbSIPK (for salicylic acid-induced protein kinase) and NbWIPK (for wound-induced protein kinase) or for both genes simultaneously, demonstrating that these MAPKs are dispensable for ROS production. We further show that NbSIPK silencing is sufficient to prevent PAMP-induced gene expression but that both MAPKs are required for bacterial immunity against two virulent strains of Pseudomonas syringae and their respective nonpathogenic mutants. These results suggest that the PAMP-triggered calcium burst is upstream of separate signaling branches, one leading to MAPK activation and then gene expression and the other to ROS production. In addition, this study highlights the essential roles of NbSIPK and NbWIPK in antibacterial immunity. Unexpectedly, negative regulatory mechanisms controlling the intensity of the PAMP-triggered calcium and ROS bursts were also revealed by this work.
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Claims 9
Claim · model-written summary
Two calcium influx inhibitors suppressed the ROS burst.
- Taxon
- Nicotiana benthamiana
Nicotiana benthamianae1 - Treatment
- calcium influx inhibitorse0
- Outcome
- reactive oxygen species (ROS) burste0
- Direction
- suppressede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Two different calcium influx inhibitors suppressed the reactive oxygen species (ROS) burst”
e1“Nicotiana benthamiana”
Claim · model-written summary
Calcium influx inhibitors suppressed MAPK activation and PAMP induced gene expression.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- calcium influx inhibitorse0
- Outcome
- activation of the mitogen-activated protein kinases (MAPKs) and PAMP-induced gene expressione0
- Direction
- suppressede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Two different calcium influx inhibitors suppressed the reactive oxygen species (ROS) burst: activation of the mitogen-activated protein kinases (MAPKs) and PAMP-induced gene expression”
e1“Nicotiana benthamiana”
Claim · model-written summary
Calcium burst remained unaffected after silencing components of ROS generation or PAMP activated MAPKs.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- plants specifically silenced for components involved in ROS generation or for MAPKs activated by PAMP treatmente0
- Outcome
- calcium burste0
- Direction
- unaffectede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The calcium burst was unaffected in plants specifically silenced for components involved in ROS generation or for MAPKs activated by PAMP treatment”
e1“Nicotiana benthamiana”
Claim · model-written summary
ROS burst persisted in plants silenced for NbSIPK.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- plants silenced for the two major defense-associated MAPK genes NbSIPKe0
- Gene
- NbSIPKe0
- Outcome
- ROS burste0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The ROS burst still occurred in plants silenced for the two major defense-associated MAPK genes NbSIPK (for salicylic acid-induced protein kinase) and NbWIPK (for wound-induced protein kinase) or for both genes simultaneously”
e1“Nicotiana benthamiana”
Claim · model-written summary
ROS burst persisted in plants silenced for NbWIPK.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- plants silenced for the two major defense-associated MAPK genes NbSIPK (for salicylic acid-induced protein kinase) and NbWIPKe0
- Gene
- NbWIPKe0
- Outcome
- ROS burste0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The ROS burst still occurred in plants silenced for the two major defense-associated MAPK genes NbSIPK (for salicylic acid-induced protein kinase) and NbWIPK (for wound-induced protein kinase) or for both genes simultaneously”
e1“Nicotiana benthamiana”
Claim · model-written summary
NbSIPK silencing prevented PAMP induced gene expression.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Treatment
- NbSIPK silencinge0
- Gene
- NbSIPKe0
- Outcome
- PAMP-induced gene expressione0
- Direction
- prevente0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“NbSIPK silencing is sufficient to prevent PAMP-induced gene expression”
e1“Nicotiana benthamiana”
Claim · model-written summary
Both MAPKs were required for immunity against virulent Pseudomonas syringae and nonpathogenic mutants.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Interactor
- Pseudomonas syringae
Pseudomonas syringaee0pathogen - Gene
- MAPKse0
- Outcome
- bacterial immunitye0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“both MAPKs are required for bacterial immunity against two virulent strains of Pseudomonas syringae and their respective nonpathogenic mutants”
e1“Nicotiana benthamiana”
Claim · model-written summary
Authors propose the PAMP triggered calcium burst precedes separate MAPK and ROS signaling branches.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Outcome
- separate signaling branchese0
- Qualifier
- PAMP-triggered calcium burste0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“PAMP-triggered calcium burst is upstream of separate signaling branches, one leading to MAPK activation and then gene expression and the other to ROS production”
e1“Nicotiana benthamiana”
Claim · model-written summary
The study revealed negative regulation of PAMP triggered calcium and ROS burst intensity.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae1
- Outcome
- intensity of the PAMP-triggered calcium and ROS burstse0
- Qualifier
- negative regulatory mechanismse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“negative regulatory mechanisms controlling the intensity of the PAMP-triggered calcium and ROS bursts were also revealed”
e1“Nicotiana benthamiana”
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loss of function · molecular: expression · Nicotiana benthamianaPositive and negative results
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