Simultaneous visualization of callose deposition and plasma membrane for live-cell imaging in plants

Masaki Kohari, Naoto Shibuya, Hanae Kaku

Research output: Contribution to journalArticlepeer-review

Abstract

Key message: The appropriate combination of fluorescent probes enabled the simultaneous visualization of callose deposition and plasma membrane in living Arabidopsis and can be useful for the cell biological study of papilla formation in plants. Abstract: Localized callose deposition at the site of fungal infection is a central part of papilla formation, which creates a barrier between the host plasma membrane and the cell wall and plays an important role in preventing the penetration of fungal hyphae into the host cells. Using chitin-induced callose deposition as a model system, we examined suitable conditions for the simultaneous visualization of callose deposition and plasma membrane dynamics in living Arabidopsis cotyledons. We found that aniline blue fluorochrome (ABF) for callose staining selectively interferes with FM dyes for membrane visualization depending on the structure of the latter compounds and the proper combination of these fluorescent dyes and staining conditions is a key for successful live-cell imaging. The established conditions enabled the live-cell imaging of chitin-induced callose deposition and host membrane systems. The established system/conditions would also be useful for the cell biological studies on the localized callose deposition in other stress/development-associated processes. The finding that the slight difference in the structure of FM dyes affects the interaction with another fluorescent dye, ABF, would also give useful suggestions for the studies where multiple fluorescent dyes are utilized for live-cell imaging.

Original languageEnglish
Pages (from-to)1517-1523
Number of pages7
JournalPlant Cell Reports
Volume39
Issue number11
DOIs
Publication statusPublished - 1 Nov 2020

Keywords

  • ABF
  • Arabidopsis
  • Callose deposition
  • Chitin oligosaccharide
  • FM dyes
  • Live-cell imaging

Fingerprint Dive into the research topics of 'Simultaneous visualization of callose deposition and plasma membrane for live-cell imaging in plants'. Together they form a unique fingerprint.

Cite this