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
[Research article]
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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. | ||||||
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Title: | Impact of salt stress, cell death, and autophagy on peroxisomes: quantitative and morphological analyses using small fluorescent probe N-BODIPY | ||||||
Series Name/Journal: | Scientific Reports | ||||||
Year of publishing : | 2017 | ||||||
Volume: | 7 | ||||||
Page range: | 1-17 | ||||||
Number of Pages: | 17 | ||||||
Publisher: | Springer Nature | ||||||
ISSN: | 2045-2322 | ||||||
Language: | English | ||||||
Publication Type: | Research article | ||||||
Refereed: | Yes | ||||||
Article category: | Scientific peer reviewed | ||||||
Version: | Published version | ||||||
Copyright: | Creative Commons: Attribution 4.0 | ||||||
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: |
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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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