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Chinese Bulletin of Botany >

2025 , Vol. 60 >Issue 1: 90 - 100

DOI: https://doi.org/10.11983/CBB24040

TECHNIQUES AND METHODS

Quantitative Analysis of Plasma Membrane Order in Live Plant Cells

  • Xiuxiu Chen ,
  • Ling Tang ,
  • Wenjia Hu ,
  • Zhaolin Yang ,
  • Xin Deng ,
  • Xiaohua Wang
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  • 1State Key Laboratory of Plant Diversity and Special Crops, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
    2China National Botanical Garden, Beijing 100093, China
    3University of Chinese Academy of Sciences, Beijing 100049, China

Received date: 2024-03-11

  Accepted date: 2024-05-27

  Online published: 2024-05-30

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Abstract

Membrane microdomains, which are highly dynamic structures rich in sterols and sphingolipids on the plasma membrane, play crucial roles in various biological processes such as signal transduction, vesicle transport, endocytosis, and exocytosis. Consequently, the investigation of membrane microdomain dynamics is an important area of research in plant cell biology. Fluorescence probes combined with fluorescence microscopy are widely used to monitor the status of living plant cells. The PA probe (push-pull pyrene) is a novel, highly efficient and stable fluorescence probe based on pyrene: however, its application in imaging studies of living plant cells is limited. In this study, we used PA probes and laser scanning confocal microscopy, combined with image processing and the polar normalized value mapping method, to quantitatively analyze the order of the plasma membrane in Arabidopsis root cells. The results revealed that the emission spectrum of the liquid-ordered phase in the plasma membrane of Arabidopsis root cells labeled with the PA probe ranged from 500-550 nm, whereas the emission spectrum of the liquid-disordered phase ranged from 580-700 nm. Treatment of wild-type plants with the sterol extraction agent MβCD resulted in a decrease in plasma membrane order. In the smt2/smt3 double mutant lacking the key methyltransferase in sterol synthesis, the plasma membrane order was consistent with that of the wild-type plants after treatment with MβCD. In the smt2/smt3 mutant, the plasma membrane order of the root hair cells was lower than the plasma membrane order of the wild-type root hair cells, indicating that sterols, as key components of membrane microdomains, play an important role in regulating the order of the plasma membrane. This study provides a straightforward and rapid detection method for monitoring the dynamic characteristics of living plant cell membranes and changes in membrane microdomains.

Key words： PA probe; phytosterol; membrane microdomains; live-cell fluorescence imaging; membrane order

Cite this article

Xiuxiu Chen , Ling Tang , Wenjia Hu , Zhaolin Yang , Xin Deng , Xiaohua Wang . Quantitative Analysis of Plasma Membrane Order in Live Plant Cells[J]. Chinese Bulletin of Botany, 2025 , 60(1) : 90 -100 . DOI: 10.11983/CBB24040

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