(*contributed equally to this work) Published: Vol 5, Iss 10, May 20, 2015 DOI: 10.21769/BioProtoc.1470 Views: 10686
Reviewed by: Maria SinetovaAnonymous reviewer(s)
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Abstract
Accumulation of metals in plant tissues, and occasionally, different cells of the same tissue, may be highly non-uniform (Seregin and Kozhevnikova, 2008). Easy-to-use histochemical methods may greatly help to investigate the distribution and accumulation of metals within and among plant tissues, and also provide information on their subcellular localization (Seregin and Kozhevnikova, 2011). The histochemical techniques of zinc (Zn) visualization are based on the formation of the blue-colored complex of Zn with the metallochrome indicator Zincon (C20H15N4NaO6S), or the green-fluorescent complex with Zinpyr-1 (C46H36Cl2N6O5) (Seregin et al., 2011; Seregin and Kozhevnikova, 2011). A method for histochemical Zn detection in plant tissues using Zinpyr-1 was first proposed by Sinclair et al. (2007), and later modified by Seregin et al. (2011), and Seregin and Kozhevnikova (2011). Histochemical data supplement the results of quantitative analysis, thus allowing a detailed study of the distribution, accumulation, and translocation pathways of Zn within the plant, which are important topics in modern plant physiology. These histochemical techniques have been successfully applied in different plant species, for example Zea mays (Seregin et al., 2011), Noccaea caerulescens and Thlaspi arvense (Kozhevnikova et al., 2014a), Capsella bursa-pastoris and Lepidium ruderale (Kozhevnikova et al., 2014b), in which Zn was detected in different root and shoot tissues. Here, we present the full staining protocols for these methods, developed or modified in our lab (Seregin and Kozhevnikova, 2011; Kozhevnikova et al., 2014a; Kozhevnikova et al., 2014b).
Keywords: Zinc localizationMaterials and Reagents
Equipment
Procedure
Representative data
Figure 1. Zn localization in plant tissues using Zinpyr-1 (A-C) and Zincon (D-G). A. Root section, B. Root, C. Leaf petiole section of Capsella bursa-pastoris exposed to 20 µM Zn for 8 weeks; D. Root section, E-F. Leaf sections, G. Leaf epidermal peel of Noccaea caerulescens exposed to 800 µM Zn for 8 weeks. Green fluorescence indicates the location of the Zn-Zinpyr-1 complex, blue colour indicates the location of the Zn-Zincon complex. Designations: C-cortex; E-endodermis; Ep-epidermis; GC-stomata guard cells; M-mesophyll; P-pericycle; Ph-phloem; R-rhizodermis; RH-root hairs; SC-subsidiary cell; VB-vascular bundle; WSC-water-storage epidermal cell; X-xylem. Bar: 10 µm (A, D-G); 50 µm (B, C)
Notes
Recipes
Acknowledgments
This work was supported by the Russian Foundation for Basic Research (RFBR, # 11-04-00513, 15-04-02236). We thank Prof. V. B. Ivanov and Dr. A. Voronkov for fruitful discussions. The protocol for Zn staining with Zinpyr-1 was modified from the work of Dr. S. A. Sinclair and co-authors (2007).
References
Article Information
Copyright
© 2015 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Seregin, I., Kozhevnikova, A. and Schat, H. (2015). Histochemical Detection of Zn in Plant Tissues. Bio-protocol 5(10): e1470. DOI: 10.21769/BioProtoc.1470.
Category
Plant Science > Plant cell biology > Tissue analysis
Plant Science > Plant physiology > Ion analysis
Cell Biology > Cell staining > Other compound
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