The plant cell wall is a complex 3D network composed of polysaccharides, lignin and proteins. The knowledge of the structure and content of each cell wall polymer is a prerequisite to understand their functions during plant development and adaptation but also to optimise their industrial applications. The analysis of cell wall compounds is complicated by their multiple molecular interactions. In this review, we present numerous methods to purify, characterise and quantify proteins, polysaccharides and lignin from the wall. Two kinds of approaches are detailed: the first presents in vitro methods which involve the breakdown of the molecular linkages between polymers thanking to chemical, physical and/or enzymatic treatments. The second approach describes in situ methods that allow the cell wall polymer characterisation thanking to many analytical techniques coupled with microscopy. If microscopy is the common point of all of them, their development is associated with improvement of analytical techniques, increasing their power of resolution.
The wood polysaccharide composition, a new analytical method, based on ionic liquid dissolution of low amount of biomass coupled with an ELISA essay of polysaccharides. In the present work, we synthesized and tested several imidazolium and 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) based ILs for their ability to solubilize Douglas-fir wood while preserving the wall polymer integrity. The couple times-temperatures have been essayed for wood dissolution. Then their efficiency for wood biomass dissolution was compared to the impact of IL on storing and/or destroy polysaccharides. Thanks to the ELISA technique with a set of mAbs against epitopes of the main hemicellulose, pectin, and protein families of cell wall components. Wood destructuration at 80˚C with the 1-ethyl-3-methylimidazolium bromide represents a good compromise of wood dissolution efficiency and low polysaccharide destruction.
We present here a new cell wall polysaccharide analysis method based on the fast and efficient ionic liquid solubilization of wood samples followed by direct immunological detections of their polysaccharide contents. To reduce both the time and the temperature of the analysis, wood powder samples are first totally disrupted in 1-ethyl-3-methylimidazolium under controlled microwave irradiation. The solubilized extracts are then directly spotted onto nitrocellulose membranes in order to specifically detect their polysaccharide contents with a set of monoclonal antibodies. Compared to the long sequential method classically used for biomass analysis, the technique described in this article allows the simultaneous treatments of higher numbers of smaller samples. It represents an interesting toolkit to rapidly screen various biomass resources for their polysaccharide contents.
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