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Protocols relating to the Structure and Composition of Plant Cell Walls

People have already exploited the physical properties of plant cell walls in, for example, the production of paper, textiles and building materials. In the future it may be possible to engineer plants to produce other environmentally friendly / biocompatible materials with different performance characteristics.

In addition, plant cell walls are potentially a very large source of carbohydrates for biofuels, and methods to better understand the ultrastructural and chemical barriers to their digestibility, as well as to monitor responses to different experimental treatments, would be very valuable in this research.

Over the past month, we have published five protocols that relate to plant cell walls: their structure and composition, and the biochemistry underlying their development. I have created a webpage where these, and any future protocols that we might publish on this subject, can be collated.

Protocols relating to Plant Cell Walls

In a sense this series on plant cell walls is a return home. The department in which I did the Cell Biology part of my undergraduate degree was very much focussed on plant biology, and TEM of recalcitrant seeds was part of the course. There was, therefore, a time when the walls of my room were littered with little printouts showing images of cells walls and sections of endoplasmic reticulum that had patterns that somehow pleased me. I had a hand-wavey “understanding” of how cell walls formed, and some sort of feeling for the fact that this was not a trivial process.

Admittedly all of this was rather a long time ago, and I suppose is only interesting if you happen to think of me solely as a chemistry-type!

It does, however, give me the small amount of credibility to enable me to say that almost everything that I learnt at University in this subject area is now out of date. In fact, it is almost completely pointless for me to mentally refer back to my undergraduate biology courses as a source of information or inspiration. This is not really because the information that that I was taught was wrong, but that the mental conclusions I made were based on an incomplete picture. This can be exemplified by the fact that there were two things about the title of the following article by Nottburger Gierlinger and Manfred Schwanninger –

Chemical Imaging of Poplar Wood Cell Walls by Confocal Raman Microscopy

– that somehow surprised me:

– There is something about the chemistry of plant cell walls that people are finding very interesting
– Raman spectroscopy has made the transition to being a microscopy technique

If people are looking at plant cell walls using confocal Raman microscopy (which is surely on the super-cool end of the techniques spectrum), they must be looking at it using other more familiar techniques as well.

Further investigation convinced me it would be a good idea to commission a collection of protocols covering a variety of approaches that can be taken to gain a better understanding of plant cells walls. I got in touch in Professor Geoffrey Fincher, and I am very grateful for his help in shepherding the preparation of four additional protocols. I would also very much like to thank the authors for all their hard work and patience; I am very pleased with these protocols and hope that they will be interesting and useful to other researchers.

These are the five protocols that resulted from this process:

Whole Plant Cell Wall Characterization using Solution-state 2D-NMR
Shawn D. Mansfield, Hoon Kim, Fachuang Lu, and John Ralph
https://www.nature.com/nprot/journal/v7/n9/full/nprot.2012.064.html

Determining the polysaccharide composition of plant cell walls
Filomena A Pettolino, Cherie Walsh, Geoffrey B Fincher & Antony Bacic
https://www.nature.com/nprot/journal/v7/n9/full/nprot.2012.081.html

Radiometric and spectrophotometric in vitro assays of glycosyltransferases involved in plant cell wall carbohydrate biosynthesis
Christian Brown, Felicia Leijon & Vincent Bulone
https://www.nature.com/nprot/journal/v7/n9/full/nprot.2012.089.html

Preparation of plant cells for transmission electron microscopy to optimize immunogold labeling of carbohydrate and protein epitopes
Sarah M Wilson & Antony Bacic
https://www.nature.com/nprot/journal/v7/n9/full/nprot.2012.096.html

Imaging of plant cell walls by confocal Raman microscopy
Notburga Gierlinger, Tobias Keplinger & Michael Harrington
https://www.nature.com/nprot/journal/v7/n9/full/nprot.2012.092.html

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