High‐throughput mapping of cell‐wall polymers within and between plants using novel microarrays

Abstract

SummaryWe describe here a methodology that enables the occurrence of cell-wall glycans to be systematically mapped throughout plants in a semi-quantitative high-throughput fashion. The technique (comprehensive microarray polymer profiling, or CoMPP) integrates the sequential extraction of glycans from multiple organs or tissues with the generation of microarrays, which are probed with monoclonal antibodies (mAbs) or carbohydratebinding modules (CBMs) with specificities for cell-wall components. The profiles generated provide a global snapshot of cell-wall composition, and also allow comparative analysis of mutant and wild-type plants, as demonstrated here for the Arabidopsis thaliana mutants fra8, mur1 and mur3. CoMPP was also applied to Physcomitrella patens cell walls and was validated by carbohydrate linkage analysis. These data provide new insights into the structure and functions of plant cell walls, and demonstrate the potential of CoMPP as a component of systems-based approaches to cell-wall biology.

Understanding plant cell-wall remodelling during the symbiotic interaction between Tuber melanosporum and Corylus avellana using a carbohydrate microarray

“…1). The CoMPP profile of the non-mycorrhizal roots points to a typical cell wall pattern for angiosperms (Moller et al 2007;Fangel et al 2012), while a decrease in all of the cell wall polymers has been revealed in the ectomycorrhizal sample, except for (1 → 3)-β-d-glucan, which can be assigned to the fungus, which also contains this polymer (Balestrini et al 2012;Fig. 2).…”

“…CoMPP was carried out as described in Moller et al (2007). Cell wall polymers were extracted sequentially from 10 mg of AIR with 300 μl of 50 mM diamino-cyclo-hexane-tetra-acetic acid (CDTA), pH 7.5 and 4 M NaOH with 0.1 % v/v NaBH4 and spotted on a nitrocellulose membrane with a pore size of 0.45 μm (Whatman, Maidstone, UK) using an Arrayjet Sprint (Arrayjey, Roslin, UK).…”

“…To investigate the changes related to root colonisation by the symbiotic fungus at the biochemical level, CoMPP (Moller et al 2007) was used to measure the intensity with which antibodies recognize specific epitopes in the cell walls of ectomycorrhizal and non-mycorrhizal root tips (Fig. 1).…”

“…Several conventional biochemical techniques are available, but carbohydrate microarrays have emerged as a useful tool for high throughput semi-quantitative analysis. One technique, known as Comprehensive Microarray Polymer Profiling (CoMPP), has been used extensively to map defined glycan structures (epitopes) across large sample sets (Moller et al 2007;Sørensen and Willats 2011). CoMPP involves the sequential extraction of cell wall polymers, which are printed as microarrays and probed with sets of cell wall-specific monoclonal antibodies (mAbs) and carbohydrate-binding modules (CBMs) (Moller et al 2007(Moller et al , 2008.…”

“…One technique, known as Comprehensive Microarray Polymer Profiling (CoMPP), has been used extensively to map defined glycan structures (epitopes) across large sample sets (Moller et al 2007;Sørensen and Willats 2011). CoMPP involves the sequential extraction of cell wall polymers, which are printed as microarrays and probed with sets of cell wall-specific monoclonal antibodies (mAbs) and carbohydrate-binding modules (CBMs) (Moller et al 2007(Moller et al , 2008. CoMPP combines the high throughput capacity of microarray technology with the specificity of mAbs and CBMs, and allows glycan profiles to be rapidly generated in diverse systems.…”