Aequorin-based luminescence imaging reveals differential calcium signalling responses to salt and reactive oxygen species in rice roots

J Exp Bot. 2015 May;66(9):2535-45. doi: 10.1093/jxb/erv043. Epub 2015 Mar 9.

Abstract

It is well established that both salt and reactive oxygen species (ROS) stresses are able to increase the concentration of cytosolic free Ca(2+) ([Ca(2+)]i), which is caused by the flux of calcium (Ca(2+)). However, the differences between these two processes are largely unknown. Here, we introduced recombinant aequorin into rice (Oryza sativa) and examined the change in [Ca(2+)]i in response to salt and ROS stresses. The transgenic rice harbouring aequorin showed strong luminescence in roots when treated with exogenous Ca(2+). Considering the histological differences in roots between rice and Arabidopsis, we reappraised the discharging solution, and suggested that the percentage of ethanol should be 25%. Different concentrations of NaCl induced immediate [Ca(2+)]i spikes with the same durations and phases. In contrast, H₂O₂ induced delayed [Ca(2+)]i spikes with different peaks according to the concentrations of H₂O₂. According to the Ca(2+) inhibitor research, we also showed that the sources of Ca(2+) induced by NaCl and H₂O₂ are different. Furthermore, we evaluated the contribution of [Ca(2+)]i responses in the NaCl- and H₂O₂-induced gene expressions respectively, and present a Ca(2+)- and H₂O₂-mediated molecular signalling model for the initial response to NaCl in rice.

Keywords: Aequorin; Oryza sativa; calcium; reactive oxygen species; rice; root; salt..

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aequorin / analysis
  • Aequorin / metabolism
  • Apoproteins / analysis
  • Apoproteins / metabolism
  • Calcium Signaling*
  • Gene Expression Regulation, Plant*
  • Oryza / genetics
  • Oryza / metabolism*
  • Plant Proteins / analysis
  • Plant Proteins / metabolism
  • Plant Roots / genetics
  • Plant Roots / metabolism
  • Plants, Genetically Modified / metabolism
  • Reactive Oxygen Species / metabolism*
  • Recombinant Proteins / analysis
  • Recombinant Proteins / metabolism
  • Sodium Chloride / metabolism*

Substances

  • Apoproteins
  • Plant Proteins
  • Reactive Oxygen Species
  • Recombinant Proteins
  • apoaequorin
  • Sodium Chloride
  • Aequorin