Probing the molecular architecture of Arabidopsis thaliana secondary cell walls using two- and three-dimensional (13)C solid state nuclear magnetic resonance spectroscopy.
The plant secondary cell wall is a thickened polysaccharide and phenolic structure, providing mechanical strength to cells, particularly in woody tissues. It is the main feedstock for the developing bioenergy and green chemistry industries. Despite the role that molecular architecture (the arrangement of biopolymers relative to each other, and their conformations) plays in dictating biomass properties, such as recalcitrance to breakdown, it is poorly understood. Here, unprocessed dry (13)C-labeled stems from the model plant Arabidopsis thaliana were analyzed by a variety of (13)C solid state magic angle spinning nuclear magnetic resonance methods, such as one-dimensional cross-polarization and direct polarization, two-dimensional refocused INADEQUATE, RFDR, PDSD, and three-dimensional DARR, demonstrating their viability for the study of native polymer arrangements in intact secondary cell walls. All carbon sites of the two main glucose environments in cellulose (previously assigned to microfibril surface and interior residues) are clearly resolved, as are carbon sites of the other major components of the secondary cell wall: xylan and lignin. The xylan carbon 4 chemical shift is markedly different from that reported previously for solution or primary cell wall xylan, indicating significant changes in the helical conformation in these dried stems. Furthermore, the shift span indicates that xylan adopts a wide range of conformations in this material, with very little in the 31 conformation typical of xylan in solution. Additionally, spatial connections of noncarbohydrate species were observed with both cellulose peaks conventionally assigned as "surface" and as "interior" cellulose environments, raising questions about the origin of these two cellulose signals.
†Department of Physics, University of Warwick, Coventry CV4 7AL, U.K.
Ray
R
Dupree
‡Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QW, U.K.
Thomas J
TJ
Simmons
‡Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QW, U.K.
Jennifer C
JC
Mortimer
†Department of Physics, University of Warwick, Coventry CV4 7AL, U.K.|‡Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QW, U.K.
Dharmesh
D
Patel
†Department of Physics, University of Warwick, Coventry CV4 7AL, U.K.
Dinu
D
Iuga
†Department of Physics, University of Warwick, Coventry CV4 7AL, U.K.
Steven P
SP
Brown
‡Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QW, U.K.
Paul
P
Dupree
D017360:Arabidopsis / Q000737:chemistry*; D029681:Arabidopsis Proteins / Q000737:chemistry; D002473:Cell Wall / Q000737:chemistry; D002482:Cellulose / Q000737:chemistry; D008031:Lignin / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D010368:Pectins / Q000737:chemistry; D018547:Plant Stems / Q000737:chemistry; D014990:Xylans / Q000737:chemistry
D029681:Arabidopsis Proteins; D014990:Xylans; D010368:Pectins; D002482:Cellulose; D008031:Lignin