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An updated protocol for high throughput plant tissue sectioning

Atkinson, Jonathan A.; Wells, Darren M.

Authors

Jonathan A. Atkinson

Darren M. Wells



Abstract

Quantification of the tissue and cellular structure of plant material is essential for the study of a variety of plant sciences applications. Currently, many methods for sectioning plant material are either low throughput or involve free-hand sectioning which requires a significant amount of practice. Here, we present an updated method to provide rapid and high-quality cross sections, primarily of root tissue but which can also be readily applied to other tissues such as leaves or stems. To increase the throughput of traditional agarose embedding and sectioning, custom designed 3D printed molds were utilized to embed 5–15 roots in a block for sectioning in a single cut. A single fluorescent stain in combination with laser scanning confocal microscopy was used to obtain high quality images of thick sections. The provided CAD files allow production of the embedding molds described here from a number of online 3D printing services. Although originally developed for roots, this method provides rapid, high quality cross sections of many plant tissue types, making it suitable for use in forward genetic screens for differences in specific cell structures or developmental changes. To demonstrate the utility of the technique, the two parent lines of the wheat (Triticum aestivum) Chinese Spring × Paragon doubled haploid mapping population were phenotyped for root anatomical differences. Significant differences in adventitious cross section area, stele area, xylem, phloem, metaxylem, and cortical cell file count were found.

Journal Article Type Article
Publication Date Oct 4, 2017
Journal Frontiers in Plant Science
Electronic ISSN 1664-462X
Publisher Frontiers Media
Peer Reviewed Peer Reviewed
Volume 8
APA6 Citation Atkinson, J. A., & Wells, D. M. (2017). An updated protocol for high throughput plant tissue sectioning. Frontiers in Plant Science, 8, https://doi.org/10.3389/fpls.2017.01721
DOI https://doi.org/10.3389/fpls.2017.01721
Keywords Tissue sectioning, root anatomy, cross section, confocal microscopy, 3D printing
Publisher URL https://www.frontiersin.org/articles/10.3389/fpls.2017.01721/full
Copyright Statement Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0

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Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0





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