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

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...

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Autores principales: Gao, Wei, Nan, Tiegui, Tan, Guiyu, Zhao, Hongwei, Tan, Weiming, Meng, Fanyun, Li, Zhaohu, Li, Qing X., Wang, Baomin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4420502/
https://www.ncbi.nlm.nih.gov/pubmed/25941807
http://dx.doi.org/10.1371/journal.pone.0123779
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author Gao, Wei
Nan, Tiegui
Tan, Guiyu
Zhao, Hongwei
Tan, Weiming
Meng, Fanyun
Li, Zhaohu
Li, Qing X.
Wang, Baomin
author_facet Gao, Wei
Nan, Tiegui
Tan, Guiyu
Zhao, Hongwei
Tan, Weiming
Meng, Fanyun
Li, Zhaohu
Li, Qing X.
Wang, Baomin
author_sort Gao, Wei
collection PubMed
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 (Cd(2+)) was developed for the first time for the wheat Triticum aestivum grown in Cd(2+)-fortified soils. Also, 1-(4-Isothiocyanobenzyl)-ethylenediamine-N,N,N,N-tetraacetic acid (ITCB-EDTA) was used to chelate the mobile Cd(2+). The ITCB-EDTA/Cd(2+) complex was fixed with proteins in situ via the isothiocyano group. A new Cd(2+)-EDTA specific monoclonal antibody, 4F3B6D9A1, was used to locate the Cd(2+)-EDTA protein complex. After staining, the fluorescence intensities of sections of Cd(2+)-positive roots were compared with those of Cd(2+)-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 Cd(2+) content in plant tissues and illustrate Cd(2+) translocation and cellular and subcellular responses of T. aestivum to Cd(2+) 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 Cd(2+) was also confirmed with energy-dispersive X-ray microanalysis. The IHC method is suitable for locating and quantifying Cd(2+) in plant tissues and can be extended to other heavy metallic ions.
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spelling pubmed-44205022015-05-12 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 PLoS One Research Article 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 (Cd(2+)) was developed for the first time for the wheat Triticum aestivum grown in Cd(2+)-fortified soils. Also, 1-(4-Isothiocyanobenzyl)-ethylenediamine-N,N,N,N-tetraacetic acid (ITCB-EDTA) was used to chelate the mobile Cd(2+). The ITCB-EDTA/Cd(2+) complex was fixed with proteins in situ via the isothiocyano group. A new Cd(2+)-EDTA specific monoclonal antibody, 4F3B6D9A1, was used to locate the Cd(2+)-EDTA protein complex. After staining, the fluorescence intensities of sections of Cd(2+)-positive roots were compared with those of Cd(2+)-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 Cd(2+) content in plant tissues and illustrate Cd(2+) translocation and cellular and subcellular responses of T. aestivum to Cd(2+) 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 Cd(2+) was also confirmed with energy-dispersive X-ray microanalysis. The IHC method is suitable for locating and quantifying Cd(2+) in plant tissues and can be extended to other heavy metallic ions. Public Library of Science 2015-05-05 /pmc/articles/PMC4420502/ /pubmed/25941807 http://dx.doi.org/10.1371/journal.pone.0123779 Text en © 2015 Gao et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Gao, Wei
Nan, Tiegui
Tan, Guiyu
Zhao, Hongwei
Tan, Weiming
Meng, Fanyun
Li, Zhaohu
Li, Qing X.
Wang, Baomin
Cellular and Subcellular Immunohistochemical Localization and Quantification of Cadmium Ions in Wheat (Triticum aestivum)
title Cellular and Subcellular Immunohistochemical Localization and Quantification of Cadmium Ions in Wheat (Triticum aestivum)
title_full Cellular and Subcellular Immunohistochemical Localization and Quantification of Cadmium Ions in Wheat (Triticum aestivum)
title_fullStr Cellular and Subcellular Immunohistochemical Localization and Quantification of Cadmium Ions in Wheat (Triticum aestivum)
title_full_unstemmed Cellular and Subcellular Immunohistochemical Localization and Quantification of Cadmium Ions in Wheat (Triticum aestivum)
title_short Cellular and Subcellular Immunohistochemical Localization and Quantification of Cadmium Ions in Wheat (Triticum aestivum)
title_sort cellular and subcellular immunohistochemical localization and quantification of cadmium ions in wheat (triticum aestivum)
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4420502/
https://www.ncbi.nlm.nih.gov/pubmed/25941807
http://dx.doi.org/10.1371/journal.pone.0123779
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