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

Gao, Wei; Nan, Tiegui; Tan, Guiyu; Zhao, Hongwei; Tan, Weiming; Meng, Fanyun; Li, Zhaohu; Li, Qing X.; Wang, Baomin

doi:10.1371/journal.pone.0123779

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

May 05 2015
By Gao, Wei ; Nan, Tiegui ; Tan, Guiyu ; ...
http://dx.doi.org/10.1371/journal.pone.0123779
Source: PLoS One. 2015; 10(5).

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Alternative Title:

PLoS One
Personal Author:

Gao, Wei ; Nan, Tiegui ; Tan, Guiyu ; Zhao, Hongwei ; Tan, Weiming ; ... More +

Gao, Wei ; Nan, Tiegui ; Tan, Guiyu ; Zhao, Hongwei ; Tan, Weiming ; Meng, Fanyun ; Li, Zhaohu ; Li, Qing X. ; Wang, Baomin Less -
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:

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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

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 Less -
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CDC Public Access
Main Document Checksum:

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urn:sha256:1b3ad47e29eeb9d9de5b914327381ebd8c98d4cfade91612b2840049865fc47e
Download URL:

https://stacks.cdc.gov/view/cdc/32374/cdc_32374_DS1.pdf
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