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Title: Subdiffraction-resolution live-cell imaging for visualizing thylakoid membranes

Journal Article · · The Plant Journal
DOI:https://doi.org/10.1111/tpj.14021· OSTI ID:1560876
ORCiD logo [1];  [2];  [1]
  1. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Biophysics and Integrated Bioimaging Div.; Univ. of California, Berkeley, CA (United States). Dept. of Plant and Microbial Biology, Howard Hughes Medical Institute
  2. Univ. of California, Berkeley, CA (United States). Dept. of Plant and Microbial Biology, Howard Hughes Medical Institute

© 2018 The Authors The Plant Journal © 2018 John Wiley & Sons Ltd The chloroplast is the chlorophyll-containing organelle that produces energy through photosynthesis. Within the chloroplast is an intricate network of thylakoid membranes containing photosynthetic membrane proteins that mediate electron transport and generate chemical energy. Historically, electron microscopy (EM) has been a powerful tool for visualizing the macromolecular structure and organization of thylakoid membranes. However, an understanding of thylakoid membrane dynamics remains elusive because EM requires fixation and sectioning. To improve our knowledge of thylakoid membrane dynamics we need to consider at least two issues: (i) the live-cell imaging conditions needed to visualize active processes in vivo; and (ii) the spatial resolution required to differentiate the characteristics of thylakoid membranes. Here, we utilize three-dimensional structured illumination microscopy (3D-SIM) to explore the optimal imaging conditions for investigating the dynamics of thylakoid membranes in living plant and algal cells. We show that 3D-SIM is capable of examining broad characteristics of thylakoid structures in chloroplasts of the vascular plant Arabidopsis thaliana and distinguishing the structural differences between wild-type and mutant strains. Using 3D-SIM, we also visualize thylakoid organization in whole cells of the green alga Chlamydomonas reinhardtii. These data reveal that high light intensity changes thylakoid membrane structure in C. reinhardtii. Moreover, we observed the green alga Chromochloris zofingiensis and the moss Physcomitrella patens to show the applicability of 3D-SIM. This study demonstrates that 3D-SIM is a promising approach for studying the dynamics of thylakoid membranes in photoautotrophic organisms during photoacclimation processes.

Research Organization:
Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES); USDOE Office of Science (SC), Biological and Environmental Research (BER)
Grant/Contract Number:
SC0018301; AC02-05CH11231
OSTI ID:
1560876
Alternate ID(s):
OSTI ID: 1462279; OSTI ID: 1559159
Journal Information:
The Plant Journal, Vol. 96, Issue 1; Related Information: Figure S1. Optical sections of reconstructed three‐dimensional structured illumination microscopy images of the side view of the chloroplast isolated from wild‐type Arabidopsis thaliana leaves.Figure S2. Optical sections of reconstructed three‐dimensional structured illumination microscopy images of the side view of the chloroplast isolated from Arabidopsis thaliana stn7 stn8 mutant leaves.Figure S3. Optical sections of reconstructed three‐dimensional structured illumination microscopy images of the side view of the chloroplast isolated from Arabidopsis thaliana tap38 mutant leaves.Figure S4. Optical sections of reconstructed three‐dimensional structured illumination microscopy images of a wild‐type Chlamydomonas reinhardtii cell.Figure S5. Additional reconstructed three‐dimensional structured illumination microscopy images showing shrinkage of thylakoid membranes at the lobe region of Chlamydomonas reinhardtii chloroplasts under high‐light conditions for 1 day.; ISSN 0960-7412
Publisher:
Society for Experimental BiologyCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 20 works
Citation information provided by
Web of Science

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Cited By (5)

Regulation of Oxygenic Photosynthesis during Trophic Transitions in the Green Alga Chromochloris zofingiensis journal February 2019
Macroorganisation and flexibility of thylakoid membranes journal October 2019
Chloroplast ultrastructure in plants journal March 2019
Spatial organization of thylakoid network in higher plants journal May 2019
Hexokinase is necessary for glucose-mediated photosynthesis repression and lipid accumulation in a green alga journal September 2019

Figures / Tables (10)


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