The Arabidopsis thaliana non-specific phospholipase C2 is involved in the response to Pseudomonas syringae attack

Zuzana Krčková, Daniela Kocourková, Michal Daněk, Jitka Brouzdová, Přemysl Pejchar, Martin Janda, Igor Pokotylo, P. Ott, Olga Valentová, Jan Martinec

Research output: Contribution to journalArticle

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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 nonspecific phospholipase C protein family. The results suggest that NPC2 is involved in the response of Arabidopsis to P. Syringae attack.

Original languageEnglish
Pages (from-to)297-310
Number of pages14
JournalAnnals of Botany
Volume121
Issue number2
DOIs
Publication statusPublished - Feb 12 2018

Fingerprint

phospholipase C
Pseudomonas syringae
Arabidopsis thaliana
Arabidopsis
phosphatidylcholines
reactive oxygen species
enzyme activity
brassinolide
flagellin
diacylglycerols
silicon
Golgi apparatus
salicylic acid
aluminum
transgenic plants
quantitative polymerase chain reaction
proteins
transcription (genetics)
promoter regions
immune response

Keywords

  • Arabidopsis thaliana
  • effector-triggered immunity
  • flagellin
  • MAMP-triggered immunity
  • non-specific phospholipase C
  • phosphatidylcholine-specific phospholipase C
  • Pseudomonas syringae
  • reactive oxygen species

ASJC Scopus subject areas

  • Plant Science

Cite this

Krčková, Z., Kocourková, D., Daněk, M., Brouzdová, J., Pejchar, P., Janda, M., ... Martinec, J. (2018). The Arabidopsis thaliana non-specific phospholipase C2 is involved in the response to Pseudomonas syringae attack. Annals of Botany, 121(2), 297-310. https://doi.org/10.1093/aob/mcx160

The Arabidopsis thaliana non-specific phospholipase C2 is involved in the response to Pseudomonas syringae attack. / Krčková, Zuzana; Kocourková, Daniela; Daněk, Michal; Brouzdová, Jitka; Pejchar, Přemysl; Janda, Martin; Pokotylo, Igor; Ott, P.; Valentová, Olga; Martinec, Jan.

In: Annals of Botany, Vol. 121, No. 2, 12.02.2018, p. 297-310.

Research output: Contribution to journalArticle

Krčková, Z, Kocourková, D, Daněk, M, Brouzdová, J, Pejchar, P, Janda, M, Pokotylo, I, Ott, P, Valentová, O & Martinec, J 2018, 'The Arabidopsis thaliana non-specific phospholipase C2 is involved in the response to Pseudomonas syringae attack', Annals of Botany, vol. 121, no. 2, pp. 297-310. https://doi.org/10.1093/aob/mcx160
Krčková Z, Kocourková D, Daněk M, Brouzdová J, Pejchar P, Janda M et al. The Arabidopsis thaliana non-specific phospholipase C2 is involved in the response to Pseudomonas syringae attack. Annals of Botany. 2018 Feb 12;121(2):297-310. https://doi.org/10.1093/aob/mcx160
Krčková, Zuzana ; Kocourková, Daniela ; Daněk, Michal ; Brouzdová, Jitka ; Pejchar, Přemysl ; Janda, Martin ; Pokotylo, Igor ; Ott, P. ; Valentová, Olga ; Martinec, Jan. / The Arabidopsis thaliana non-specific phospholipase C2 is involved in the response to Pseudomonas syringae attack. In: Annals of Botany. 2018 ; Vol. 121, No. 2. pp. 297-310.
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AU - Brouzdová, Jitka

AU - Pejchar, Přemysl

AU - Janda, Martin

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