A maceration technique for soft plant tissue without hazardous chemicals

• Published in: Applications in plant sciences Vol. 11; no. 5; pp. e11543 - n/a
• Main Authors: Klahs, Phillip C., McMurchie, Elizabeth K., Nikkel, Jordan J., Clark, Lynn G.
• Format: Journal Article
• Language: English
• Published: United States John Wiley & Sons, Inc 01.09.2023; John Wiley and Sons Inc; Wiley
• Subjects: cell isolation; cost effectiveness; Dissection; Enzymes; epidermal peel; Epidermis; herbaria; Leaves; maceration; Microscopy; pectinase; plant tissues; polygalacturonase; Protocol Note; maceration; pectinase; cell isolation; dissection; epidermal peel
• ISSN: 2168-0450; 2168-0450
• DOI: 10.1002/aps3.11543

Current methods for maceration of plant tissue use hazardous chemicals. The new method described here improves the safety of dissection and maceration of soft plant tissues for microscopic imaging by using the harmless enzyme pectinase. Leaf material from a variety of land plants was obtained from living plants and dried herbarium specimens. Concentrations of aqueous pectinase and soaking schedules were optimized, and tissues were manually dissected while submerged in fresh solution following a soaking period. Most leaves required 2-4 h of soaking; however, delicate leaves could be macerated after 30 min while tougher leaves required 12 h to 3 days of soaking. Staining techniques can also be used with this method, and permanent or semi-permanent slides can be prepared. The epidermis, vascular tissue, and individual cells were imaged at magnifications of 10× to 400×. Only basic safety precautions were needed. This pectinase method is a cost-effective and safe way to obtain images of epidermal peels, separated tissues, or isolated cells from a wide range of plant taxa.