Live-cell imaging of phosphatidic acid dynamics in pollen tubes visualized by Spo20p-derived biosensor. - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue New Phytologist Année : 2014

Live-cell imaging of phosphatidic acid dynamics in pollen tubes visualized by Spo20p-derived biosensor.

Martin Potocký
  • Fonction : Auteur
Roman Pleskot
  • Fonction : Auteur
Přemysl Pejchar
  • Fonction : Auteur
Benedikt Kost
  • Fonction : Auteur
Viktor Zárský
  • Fonction : Auteur

Résumé

Although phosphatidic acid (PA) is structurally the simplest membrane phospholipid, it has been implicated in the regulation of many cellular events, including cytoskeletal dynamics, membrane trafficking and stress responses. Plant PA shows rapid turnover but the information about its spatio-temporal distribution in plant cells is missing. Here we demonstrate the use of a lipid biosensor that enables us to monitor PA dynamics in plant cells. The biosensor consists of a PA-binding domain of yeast SNARE Spo20p fused to fluorescent proteins. Live-cell imaging of PA dynamics in transiently transformed tobacco (Nicotiana tabacum) pollen tubes was performed using confocal laser scanning microscopy. In growing pollen tubes, PA shows distinct annulus-like fluorescence pattern in the plasma membrane behind the extreme tip. Coexpression studies with markers for other plasmalemma signaling lipids phosphatidylinositol 4,5-bisphosphate and diacylglycerol revealed limited colocalization at the shoulders of the apex. PA distribution and concentrations show distinct responses to various lipid signaling inhibitors. Fluorescence recovery after photobleaching (FRAP) analysis suggests high PA turnover in the plasma membrane. Our data show that a biosensor based on the Spo20p-PA binding domain is suitable for live-cell imaging of PA also in plant cells. In tobacco pollen tubes, distinct subapical PA maximum corroborates its involvement in the regulation of endocytosis and actin dynamics.

Dates et versions

hal-02613218 , version 1 (19-05-2020)

Identifiants

Citer

Martin Potocký, Roman Pleskot, Přemysl Pejchar, Nicolas Vitale, Benedikt Kost, et al.. Live-cell imaging of phosphatidic acid dynamics in pollen tubes visualized by Spo20p-derived biosensor.. New Phytologist, 2014, 203 (2), pp.483-494. ⟨10.1111/nph.12814⟩. ⟨hal-02613218⟩

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