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

Deirdre Fahy, Marwa N. M. E Sanad, Kerstin Duscha, Madison Lyons, Fuquan Liu, Peter Bozhkov, Hans-Henning Kunz, Jianping Hu, H. Ekkehard Neuhaus, Patrick G. Steel, Andrei Smertenko

Research output: Contribution to journalArticle

19 Citations (Scopus)
176 Downloads (Pure)

Abstract

Plant peroxisomes maintain a plethora of key life processes including fatty acid β-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 saltsusceptible 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.
Original languageEnglish
Article number39069
Number of pages17
JournalScientific Reports
Volume7
DOIs
Publication statusPublished - 01 Feb 2017

Keywords

  • peroxisome
  • stress
  • plant
  • N-BODIPY

Fingerprint Dive into the research topics of 'Impact of salt stress, cell death, and autophagy on peroxisomes: quantitative and morphological analyses using small fluorescent probe N-BODIPY'. Together they form a unique fingerprint.

  • Cite this

    Fahy, D., Sanad, M. N. M. E., Duscha, K., Lyons, M., Liu, F., Bozhkov, P., Kunz, H-H., Hu, J., Neuhaus, H. E., Steel, P. G., & Smertenko, A. (2017). Impact of salt stress, cell death, and autophagy on peroxisomes: quantitative and morphological analyses using small fluorescent probe N-BODIPY. Scientific Reports, 7, [39069 ]. https://doi.org/10.1038/srep39069