Detection of cytokinins and auxin in plant tissues using histochemistry and immunocytochemistry

We report a new method for histochemical localization of cytokinins (CKs) in plant tissues based on bromophenol blue/silver nitrate staining. The method was validated by immunohistochemistry using anti-trans-zeatin riboside antibody. Indole-3-acetic acid (auxin, IAA) was localized by anti-IAA antibo...

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Bibliographic Details
Published in:	Biotechnic & histochemistry Vol. 93; no. 2; p. 149
Main Authors:	Bedetti, C S, Jorge, N C, Trigueiro, Fcg, Bragança, G P, Modolo, L V, Isaias, Rms
Format:	Journal Article
Language:	English
Published:	England 01-01-2018
Subjects:	Bromphenol Blue - chemistry Cytokinins - analysis Cytokinins - chemistry Immunohistochemistry - methods Indoleacetic Acids - analysis Indoleacetic Acids - chemistry Plant Roots - chemistry Plants - chemistry Silver Nitrate - chemistry Staining and Labeling - methods cytokinins auxin histochemistry plants immunocytochemistry indole-3-acetic acid thin layer chromatography zeatin
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Summary:	We report a new method for histochemical localization of cytokinins (CKs) in plant tissues based on bromophenol blue/silver nitrate staining. The method was validated by immunohistochemistry using anti-trans-zeatin riboside antibody. Indole-3-acetic acid (auxin, IAA) was localized by anti-IAA antibody in plant tissues as a proof for IAA histolocalization. We used root sections, because they are major sites of CKs synthesis, and insect galls of Piptadenia gonoacantha that accumulate IAA. Immunostaining confirmed the presence of zeatin and sites of accumulation of IAA indicated by histochemistry. The colors developed by histochemical reactions in free-hand sections of plant tissues were similar to those obtained by thin layer chromatography (TLC), which reinforced the reactive sites of zeatin. The histochemical method for detecting CKs is useful for galls and roots, whereas IAA detection is more efficient for gall tissues. Therefore, galls constitute a useful model for validating histochemical techniques due to their rapid cell cycles and relatively high accumulation of plant hormones.
ISSN:	1473-7760
DOI:	10.1080/10520295.2017.1417640