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. 2023 Sep 18;11(5):e11543.
doi: 10.1002/aps3.11543. eCollection 2023 Sep-Oct.

A maceration technique for soft plant tissue without hazardous chemicals

Phillip C Klahs  1   2 , Elizabeth K McMurchie  1 , Jordan J Nikkel  1 , Lynn G Clark  1
Affiliations

A maceration technique for soft plant tissue without hazardous chemicals

Phillip C Klahs et al. Appl Plant Sci. .

Abstract

Premise: 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.

Methods and results: 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.

Conclusions: 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.

Keywords: cell isolation; dissection; epidermal peel; maceration; pectinase.

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Figures

Figure 1
Figure 1
Microscopic images of leaf anatomy after maceration with pectinase. (A) Vascular bundles of Euphorbia ‘Starblast Snowdrift’. Scale bar = 100 μm. (B) Petioles of Helianthus annuus ‘Teddy Bear’ with (left) and without (right) trichomes. Scale bar = 1000 μm. (C) Epidermal peel in progress showing vasculature in Rhododendron PJM Group. Scale bar = 200 μm. (D) Epidermis of Gossypium barbadense. Scale bar = 50 μm. (E) Aerenchyma of Typha angustifolia exposed after removal of epidermis. Scale bar = 1000 μm. (F) Mesophyll cells of Sempervivum cf. tectorum. Scale bar = 100 μm. (G) Epidermal cells of Thelypteris dentata. Scale bar = 50 μm. (H) Stomata and silica cells from fresh material of Phragmites australis. Scale bar = 50 μm. (I) Epidermal layer from fresh material of Pinus sylvestris being removed from leaf with a tweezer. Scale bar = 400 μm. (J) Epidermal cells of Athyrium filix‐femina from herbarium specimen. Scale bar = 50 μm. (K) Stomata and silica cells of Phragmites australis from herbarium specimen. Scale bar = 50 μm. (L) Mesophyll exposed after removal of epidermis of Pinus sylvestris from herbarium specimen. Scale bar = 400 μm.
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References

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