Maule Staining for Lignin: A Technique to Characterize Lignin Distribution in Arabidopsis Thaliana Stem Sections

Published: April 30, 2023

Abstract

Source: Pradhan Mitra, P. et al., Histochemical Staining of Arabidopsis thaliana Secondary Cell Wall Elements. J. Vis. Exp. (2014).

In this video, we describe the Maule staining technique for staining syringyl lignin present in the specialized plant cells. This staining technique helps to characterize the lignin distribution in the xylem and interfascicular fibers.

Protocol

1. Mäule Staining Dissolve 0.2 g of potassium permanganate in 40 ml distilled water to prepare a 0.5% potassium permanganate solution. Store in a dark bottle at room temperature for a maximum of 7 days. Prepare a 3.7% HCl solution by adding 1 ml concentrated HCl (37 N) to 9 ml distilled water (1:10). Prepare a fresh 3.7% HCl solution on the day of the experiment. Make sure that the 37 N HCl stock solution being used is not too old. Concentrated ammonium hydroxide solution (14.8 M) s…

Representative Results

Figure 1. Mäule staining of A. thaliana stem cross-sections. Mäule staining (red color) of a wild type A. thaliana stem section showing the normal lignin deposition in the walls of interfascicular fibers and xylem cells (A-E). The positions of the epidermis and cortex (Ep), interfascicular fibers (Fi), pith (Pi), and xylem (Xy) are indicated. Magnifications: Panel A: 5X; Panel B: 10X; Panel C: 20X; Panels D and E: 40X….

Divulgations

The authors have nothing to disclose.

Materials

Agarose   EMD MERC2125  CAS Number: 9012-36-6
Hydrochloric acid    EMD HX0603-75 CAS Number: 7647-01-0
Vibratome   Leica  Leica Vibrating blade microtome VT1000S http://www.leicabiosystems.com/ products/sectioning/vibratingblade-microtomes/details/product/ leica-vt1000-s/
Razor   American Safety razor company  Item # 60-0139-0000 Stainless Steel Double Edge Blade (Personna Super)
Screw Cap Microcentrifuge Tubes (2 ml)   VWR 16466-044
Microcentrifuge Tubes (0.6 ml)   Axygen Scientific  MCT-060-C
Tissue adhesive   Ted Pella Inc 10033 Store at 4 °C or 20 °C for 3 months or longer storage
Camera with CCD chip with no mechanical shutter  Hamamatsu  C4742-95
Camera software   Molecular Devices MetaMorph version 7.7.0.0
Imagining anaylsis   Adobe Photoshop CS4
Micro Cover Glasses, Square, No. 1  VWR  48366-067  22 x 22 mm (7/8 x 7/8")-Cover glasses are corrosion-resistant
Frosted Micro Slides, 1 mm    VWR 48312-003 75 x 25 mm – 1 mm
Ammonium hydroxide EMD   AX1303-6  CAS Number: 1336-21-6
Potassium permanganate  Sigma 223468  CAS Number 7722-64-7

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Maule Staining for Lignin: A Technique to Characterize Lignin Distribution in Arabidopsis Thaliana Stem Sections. J. Vis. Exp. (Pending Publication), e21058, doi: (2023).

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