Abstract Detail

Education and Outreach

Klahs, Phillip [1], McMurchie, Elizabeth [2], Nikkel, Jordan [3], Clark, Lynn [4].

A maceration technique for soft plant tissue without hazardous chemicals.

Soft tissue maceration techniques have been used to analyze a wide variety of anatomical features in plants and contribute to expanding our general knowledge of plant anatomy. Some structures observed with these methods can also be useful in delimiting taxonomic boundaries and provide insight into the ecology and evolutionary history of these organisms. Various procedures have been developed to isolate leaf epidermal tissue, vascular bundles, and even individual cells. However, many of these methods rely on the use of hazardous chemicals or are specific to one taxon. Scanning electron microscopy (SEM) is often used to accomplish some of these same objectives but is very costly and time-consuming. Here, we propose a protocol for a non-hazardous and inexpensive soft tissue maceration technique that allows for the analysis of a variety of anatomical features in many different plant taxa. This technique makes use of the enzyme pectinase, which breaks down pectin, the primary component of the middle lamella between the cell walls of neighboring plant cells. Pectinase is non-hazardous, inexpensive, and shelf-stable at room temperature when purchased as a powder. Plant material should be soaked in a pectinase solution made with this powder, ideally for between 1-3 hours. After this, the plant tissue can be manipulated under a stereo microscope to obtain the desired features. Staining techniques can also be utilized to aid in visualization. Plant material may then be mounted on slides for analysis using light microscopy. We tested this method on 12 different species from various lineages with representatives from across the entire plant kingdom. We have obtained and photographed isolated anatomical features from each species. These images will provide detailed views of features like the leaf epidermis, vascular tissue, and individual cells. Given that this method is inexpensive, relatively safe, and not particularly time consuming, we recommend it as a practical application for a variety of settings. Researchers can utilize this technique as a form of preliminary analysis before moving on to more expensive methods, like SEM. We also encourage educators to introduce this technique in classrooms for lessons covering biology and plant anatomy.

1 - Iowa State University, 251 Bessey Hall, 2200 Osborn Drive, Ames, IA, 50011, United States
2 - Iowa State University, Ecology, Evolution, and Organismal Biology, 2200 Osborn Drive, Ames, IA, 50011, USA
3 - Iowa State University, 2200 Osborn Drive, Ames, IA, 50011, United States
4 - Iowa State University, Department Of Ecology, Evolution, And Organismal Biology, 251 Bessey Hall, 2200 Osborn Dr., Ames, IA, 50011, United States

Keywords:
Anatomy
methods
maceration
Education
microscopy
technique.

Presentation Type: Oral Paper
Number: EO2005
Abstract ID:573
Candidate for Awards:None