Changes in cytosolic ATP levels and intracellular morphology during bacteria-induced hypersensitive cell death as revealed by real-time fluorescence microscopy imaging

Noriyuki Hatsugai, Vadim Perez Koldenkova, Hiromi Imamura, Hiroyuki Noji, Takeharu Nagai

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Hypersensitive cell death is known to involve dynamic remodeling of intracellular structures that uses energy released during ATP hydrolysis. However, the relationship between intracellular structural changes and ATP levels during hypersensitive cell death remains unclear. Here, to visualize ATP dynamics directly in real time in individual living plant cells, we applied a genetically encoded Förster resonance energy transfer (FRET)-based fluorescent ATP indicator, ATeam1.03-nD/nA, for plant cells. Intracellular ATP levels increased approximately 3 h after inoculation with the avirulent strain DC3000/avrRpm1 of Pseudomonas syringae pv. tomato (Pst), which was accompanied by the simultaneous disappearance of transvacuolar strands and appearance of bulb-like structures within the vacuolar lumen. Approximately 5 h after bacterial inoculation, the bulb-like structures disappeared and ATP levels drastically decreased. After another 2 h, the large central vacuole was disrupted. In contrast, no apparent changes in intracellular ATP levels were observed in the leaves inoculated with the virulent strain Pst DC3000. The Pst DC3000/avrRpm1-induced hypersensitive cell death was strongly suppressed by inhibiting ATP synthesis after oligomycin A application within 4 h after bacterial inoculation. When the inhibitor was applied 7 h after bacterial inoculation, cell death was unaffected. These observations show that changes in intracellular ATP levels correlate with intracellular morphological changes during hypersensitive cell death, and that ATP is required just before vacuolar rupture in response to bacterial infection.

Original languageEnglish (US)
Pages (from-to)1768-1775
Number of pages8
JournalPlant and Cell Physiology
Volume53
Issue number10
DOIs
StatePublished - Oct 2012

Bibliographical note

Funding Information:
This work was supported by the Sumitomo Foundation [to N.H.]; the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan [Grants-in-Aid for Scientific Research (grant No. 23570043 to N.H.), and the Cooperative Research Program of ‘Network Joint Research Center for Materials and Devices’ (to T.N.)].

Keywords

  • ATP
  • Disease resistance
  • FRET
  • Fluorescence bioimaging
  • Hypersensitive cell death
  • Plant immunity

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