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….
開示
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 |
