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Genomic Science Program

2006 Awardee

Streamlined Method for Biomass Whole-Cell-WallStructural Profiling

INVESTIGATOR(S): Ralph, J. (Collaborators:Lu, F; Sundberg, B.; Mansfield, S.; others)

INSTITUTION: U.S. Dairy Forage ResearchCenter, USDA-Agricultural Research Service

NON-TECHNICAL SUMMARY: In wide-rangingresearch aimed at altering plant cell wall characteristics,by conventional breeding or modern genetic methods, one ofthe biggest problems is in delineating the effects on thecell wall. Plant cell walls are a complex conglomerate of avariety of polysaccharides and lignin. Each component aloneis complex, and their interactions are only poorlycharacterized and understood. The most common approach hasbeen to isolate and purify components and to characterizethem in detail using a variety of methods. Such studieswill always be necessary. As studies in whichlignin-biosynthetic-pathway enzymes were targeted have madeabundantly clear, simple compositional analysis is notsufficient. Some plants with only minor compositionalchanges have drastically altered chemical structure thatbelies the important alterations that can be made inprocesses ranging from natural digestibility in ruminantanimals to industrial chemical pulping.

How can the structural components of the cell wall bereadily characterized? Although other methods have theirplace, and can be more rapid (e.g. NIR), the difficulty ininterpretation of some spectral methods, or the destructionof structure by chemical methods, assures that key featuresof cell walls benefiting, for example, biomass productionand conversion are lost. A promising recent approach is thedissolution of the whole cell wall and NMR analysis. Weintend to extend the methodologies to provide rapidstructural profiling of plant materials, aiming for a"screening rate" of 20-30 samples per day. Such methodologywill be useful to plant researchers worldwide.

OBJECTIVES: To provide the plant cell walland biomass research communities with improved methods forpolysaccharide and lignin structural profiling, based oncomplete cell wall solubilization and NMR. The aim is todevelop and streamline procedures to allow 20-30 samplesper day to be profiled.

APPROACH: The following improvements tothe Dissolution/NMR method will be sought: a. Provide thenecessary database, via model compounds and isolatedcomponents, to characterize component polysaccharides andlignins in whole-cell-wall mixtures. b. Optimize millingconditions for the various biomass sample types and seekalternative solutions that require less rigorous milling.c. Attempt to develop improved rapid dissolution methodsthat can be performed directly in the NMR tube; determinesolvent systems that do not interfere with the correlationcontours from polysaccharide and lignin components. d.Develop NMR methods that allow the crucial HSQC NMR spectrato be acquired in under 1 hour (on the whole cell wallsample). e. Develop methods for databasing and quantifyingthe 2D NMR cell wall spectra. f. With collaborators,attempt to develop chemometrics methods that can be appliedto 2D NMR data.

KEYWORDS: lignin; plant chemistry; plantstructure, polysaccharide, digestibility, chemical pulping,plant cell wall, NMR.


Name: Ralph, J.
Phone: 608-890-0071
Fax: 608-890-0076



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