Cellular and Subcellular Immunohistochemical Localization and Quantification of Cadmium Ions in Wheat (Triticum aestivum)

Details

• Alternative Title:
  PLoS One
• Personal Author:
  Gao, Wei ; Nan, Tiegui ; Tan, Guiyu ; Zhao, Hongwei ; Tan, Weiming ; Meng, Fanyun ; Li, Zhaohu ; Li, Qing X. ; Wang, Baomin
• Description:
  The distribution of metallic ions in plant tissues is associated with their toxicity and is important for understanding mechanisms of toxicity tolerance. A quantitative histochemical method can help advance knowledge of cellular and subcellular localization and distribution of heavy metals in plant tissues. An immunohistochemical (IHC) imaging method for cadmium ions (Cd2+) was developed for the first time for the wheat Triticum aestivum grown in Cd2+-fortified soils. Also, 1-(4-Isothiocyanobenzyl)-ethylenediamine-N,N,N,N-tetraacetic acid (ITCB-EDTA) was used to chelate the mobile Cd2+. The ITCB-EDTA/Cd2+ complex was fixed with proteins in situ via the isothiocyano group. A new Cd2+-EDTA specific monoclonal antibody, 4F3B6D9A1, was used to locate the Cd2+-EDTA protein complex. After staining, the fluorescence intensities of sections of Cd2+-positive roots were compared with those of Cd2+-negative roots under a laser confocal scanning microscope, and the location of colloidal gold particles was determined with a transmission electron microscope. The results enable quantification of the Cd2+ content in plant tissues and illustrate Cd2+ translocation and cellular and subcellular responses of T. aestivum to Cd2+ stress. Compared to the conventional metal-S coprecipitation histochemical method, this new IHC method is quantitative, more specific and has less background interference. The subcellular location of Cd2+ was also confirmed with energy-dispersive X-ray microanalysis. The IHC method is suitable for locating and quantifying Cd2+ in plant tissues and can be extended to other heavy metallic ions.
• Subjects:
  Cadmium Immunohistochemistry Microscopy, Confocal Microscopy, Electron, Transmission Triticum
• Source:
  PLoS One. 2015; 10(5).
• Pubmed ID:
  25941807
• Pubmed Central ID:
  PMC4420502
• Document Type:
  Journal Article
• Funding:
  2U54OH007550-11/OH/NIOSH CDC HHS/United States ; 5 G12 RR003061-26/RR/NCRR NIH HHS/United States ; 8 G12 MD007601-26/MD/NIMHD NIH HHS/United States ; Cancer Research UK/United Kingdom
• Volume:
  10
• Issue:
  5
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:1b3ad47e29eeb9d9de5b914327381ebd8c98d4cfade91612b2840049865fc47e
• Download URL:
  https://stacks.cdc.gov/view/cdc/32374/cdc_32374_DS1.pdf
• File Type:
  [PDF - 6.41 MB ]