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

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

1. 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.
   1. 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) should be stored at 4 °C to prevent the molarity of the saturated solution from decreasing.
2. Transfer stem sections to a 2.0 ml micro....

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), interfa.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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