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Research article - Peer-reviewed, 2017

Impact of salt stress, cell death, and autophagy on peroxisomes: quantitative and morphological analyses using small fluorescent probe N-BODIPY

Fahy, D.; Sanad, Marwa NME; Duscha, Kerstin; Lyons, Madison; Liu, Fuquan; Bozhkov, Peter; Kunz, Hans-Henning; Hu, Jianping; Neuhaus, Ekkehard; Steel, Patrick G; Smertenko, Andrei P.

Abstract

Plant peroxisomes maintain a plethora of key life processes including fatty acid beta-oxidation, photorespiration, synthesis of hormones, and homeostasis of reactive oxygen species (ROS). Abundance of peroxisomes in cells is dynamic; however mechanisms controlling peroxisome proliferation remain poorly understood because measuring peroxisome abundance is technically challenging. Counting peroxisomes in individual cells of complex organs by electron or fluorescence microscopy is expensive and time consuming. Here we present a simple technique for quantifying peroxisome abundance using the small probe Nitro-BODIPY, which in vivo fluoresces selectively inside peroxisomes. The physiological relevance of our technique was demonstrated using salinity as a known inducer of peroxisome proliferation. While significant peroxisome proliferation was observed in wildtype Arabidopsis leaves following 5-hour exposure to NaCl, no proliferation was detected in the salt-susceptible mutants fry1-6, sos1-14, and sos1-15. We also found that N-BODIPY detects aggregation of peroxisomes during final stages of programmed cell death and can be used as a marker of this stage. Furthermore, accumulation of peroxisomes in an autophagy-deficient Arabidopsis mutant atg5 correlated with N-BODIPY labeling. In conclusion, the technique reported here enables quantification of peroxisomes in plant material at various physiological settings. Its potential applications encompass identification of genes controlling peroxisome homeostasis and capturing stress-tolerant genotypes.

Keywords

peroxisomes; cell processes; cell physiology; Arabidopsis; quantitative analyses

Published in

Scientific Reports
2017, volume: 7, article number: 39068
Publisher: NATURE PUBLISHING GROUP

Authors' information

Fahy, D.
Washington State University
Sanad, Marwa NME
Washington State University
Duscha, Kerstin
University of Kaiserslautern
Lyons, Madison
Washington State University
Liu, Fuquan
Queen's University Belfast
Swedish University of Agricultural Sciences, The Department of Chemistry and Biotechnology
Kunz, Hans-Henning
Washington State University
Hu, Jianping
Michigan State University
Neuhaus, Ekkehard
University of Kaiserslautern
Steel, Patrick G
University of Durham
Smertenko, Andrei P.
Washington State University

UKÄ Subject classification

Cell Biology

Publication Identifiers

DOI: https://doi.org/10.1038/srep39069

URI (permanent link to this page)

https://res.slu.se/id/publ/80376