Abstract
Background and Aims: The non-specific phospholipase C (NPC) is a new member of the plant phospholipase family that reacts to abiotic environmental stresses, such as phosphate deficiency, high salinity, heat and aluminium toxicity, and is involved in root development, silicon distribution and brassinolide signalling. Six NPC genes (NPC1-NPC6) are found in the Arabidopsis genome. The NPC2 isoform has not been experimentally characterized so far. Methods: The Arabidopsis NPC2 isoform was cloned and heterologously expressed in Escherichia coli. NPC2 enzyme activity was determined using fluorescent phosphatidylcholine as a substrate. Tissue expression and subcellular localization were analysed using GUS- and GFP-tagged NPC2. The expression patterns of NPC2 were analysed via quantitative real-time PCR. Independent homozygous transgenic plant lines overexpressing NPC2 under the control of a 35S promoter were generated, and reactive oxygen species were measured using a luminol-based assay. Key Results: The heterologously expressed protein possessed phospholipase C activity, being able to hydrolyse phosphatidylcholine to diacylglycerol. NPC2 tagged with GFP was predominantly localized to the Golgi apparatus in Arabidopsis roots. The level of NPC2 transcript is rapidly altered during plant immune responses and correlates with the activation of multiple layers of the plant defence system. Transcription of NPC2 decreased substantially after plant infiltration with Pseudomonas syringae, flagellin peptide flg22 and salicylic acid treatments and expression of the effector molecule AvrRpm1. The decrease in NPC2 transcript levels correlated with a decrease in NPC2 enzyme activity. NPC2-overexpressing mutants showed higher reactive oxygen species production triggered by flg22. Conclusions: This first experimental characterization of NPC2 provides new insights into the role of the non-specific phospholipase C protein family. The results suggest that NPC2 is involved in the response of Arabidopsis to P. syringae attack.
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Claims 10
Claim · model-written summary
Arabidopsis NPC2 hydrolysed phosphatidylcholine to diacylglycerol, showing phospholipase C activity.
Not reportedTaxon
Verbatim
e0“The heterologously expressed protein possessed phospholipase C activity, being able to hydrolyse phosphatidylcholine to diacylglycerol.”
Claim · model-written summary
GFP-tagged NPC2 was predominantly localized to the Golgi apparatus in Arabidopsis roots.
- Taxon
- Arabidopsis
Arabidopsise0 - Organ / stage
- rootse0
- Gene
- NPC2e0
- Outcome
- localized to the Golgi apparatuse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“NPC2 tagged with GFP was predominantly localized to the Golgi apparatus in Arabidopsis roots.”
Claim · model-written summary
NPC2 transcript levels changed rapidly during plant immune responses and correlated with activation of multiple defence layers.
Not reportedTaxon
Verbatim
e0“The level of NPC2 transcript is rapidly altered during plant immune responses and correlates with the activation of multiple layers of the plant defence system.”
Claim · model-written summary
Pseudomonas syringae infiltration substantially decreased NPC2 transcription.
- Treatment
- plant infiltration with Pseudomonas syringaee0
- Interactor
- Pseudomonas syringae
Pseudomonas syringaee0pathogen - Gene
- NPC2e0
- Outcome
- Transcription of NPC2e0
- Direction
- decreasede0
Not reportedTaxonComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Transcription of NPC2 decreased substantially after plant infiltration with Pseudomonas syringae, flagellin peptide flg22 and salicylic acid treatments and expression of the effector molecule AvrRpm1.”
Claim · model-written summary
Flg22 treatment substantially decreased NPC2 transcription.
Not reportedTaxonComparator
Verbatim
e0“Transcription of NPC2 decreased substantially after plant infiltration with Pseudomonas syringae, flagellin peptide flg22 and salicylic acid treatments and expression of the effector molecule AvrRpm1.”
Claim · model-written summary
Salicylic acid treatment substantially decreased NPC2 transcription.
Not reportedTaxonComparator
Verbatim
e0“Transcription of NPC2 decreased substantially after plant infiltration with Pseudomonas syringae, flagellin peptide flg22 and salicylic acid treatments and expression of the effector molecule AvrRpm1.”
Claim · model-written summary
Expression of AvrRpm1 substantially decreased NPC2 transcription.
- Treatment
- expression of the effector molecule AvrRpm1e0
- Gene
- NPC2e0
- Outcome
- Transcription of NPC2e0
- Direction
- decreasede0
Not reportedTaxonComparator
Verbatim
e0“Transcription of NPC2 decreased substantially after plant infiltration with Pseudomonas syringae, flagellin peptide flg22 and salicylic acid treatments and expression of the effector molecule AvrRpm1.”
Claim · model-written summary
Decreased NPC2 transcription correlated with decreased NPC2 enzyme activity.
- Gene
- NPC2e0
- Outcome
- NPC2 enzyme activitye0
- Direction
- decreasee0
- Qualifier
- decrease in NPC2 transcript levelse0
Not reportedTaxon
Verbatim
e0“The decrease in NPC2 transcript levels correlated with a decrease in NPC2 enzyme activity.”
Claim · model-written summary
NPC2 overexpression increased flg22-triggered reactive oxygen species production.
- Material
- NPC2-overexpressing mutantse0
- Treatment
- flg22e0
- Outcome
- reactive oxygen species productione0
- Direction
- highere0
Not reportedTaxonComparator
Verbatim
e0“NPC2-overexpressing mutants showed higher reactive oxygen species production triggered by flg22.”
Claim · model-written summary
The authors suggest NPC2 participates in Arabidopsis response to P. syringae attack.
- Taxon
- ArabidopsisArabidopsise0
- Interactor
- P. syringaePseudomonas syringaee0pathogen
- Gene
- NPC2e0
- Outcome
- response of Arabidopsis to P. syringae attacke0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The results suggest that NPC2 is involved in the response of Arabidopsis to P. syringae attack.”
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