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

Fahy, D. and Sanad, Marwa NME and Duscha, Kerstin and Lyons, Madison and Liu, Fuquan and Bozhkov, Peter and Kunz, Hans-Henning and Hu, Jianping and Neuhaus, Ekkehard and Steel, Patrick G and Smertenko, Andrei P. (2017). Impact of salt stress, cell death, and autophagy on peroxisomes: quantitative and morphological analyses using small fluorescent probe N-BODIPY. Scientific Reports. 7, 1-17
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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.

Authors/Creators:Fahy, D. and Sanad, Marwa NME and Duscha, Kerstin and Lyons, Madison and Liu, Fuquan and Bozhkov, Peter and Kunz, Hans-Henning and Hu, Jianping and Neuhaus, Ekkehard and Steel, Patrick G and Smertenko, Andrei P.
Title:Impact of salt stress, cell death, and autophagy on peroxisomes: quantitative and morphological analyses using small fluorescent probe N-BODIPY
Series/Journal:Scientific Reports (2045-2322)
Year of publishing :2017
Volume:7
Page range:1-17
Number of Pages:17
Publisher:Springer Nature
ISSN:2045-2322
Language:English
Publication Type:Journal article
Refereed:Yes
Article category:Scientific peer reviewed
Version:Published version
Full Text Status:Public
Subjects:(A) Swedish standard research categories 2011 > 1 Natural sciences > 106 Biological Sciences (Medical to be 3 and Agricultural to be 4) > Cell Biology
Agrovoc terms:cell physiology, cell physiological processes, Arabidopsis
Keywords:peroxisomes, cell processes, cell physiology, Arabidopsis, quantitative analyses
URN:NBN:urn:nbn:se:slu:epsilon-e-4479
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-e-4479
Additional ID:
Type of IDID
DOI10.1038/srep39069
Web of Science (WoS)000393158600001
ID Code:14710
Faculty:NJ - Fakulteten för naturresurser och jordbruksvetenskap
Department:(NL, NJ) > Department of Chemistry and Biotechnology (140101-161231)
Deposited By: SLUpub Connector
Deposited On:14 Nov 2017 07:38
Metadata Last Modified:14 Nov 2017 07:38

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