Plant cell-walls methods

R99 T. J. Bootten, P. J. Harris, L. D. Melton and R. H. Newman, Using Sohd-State C Nuclear Magnetic Resonance Spectroscopy to Study the Molecular Organisation of Primary Plant Cell Walls , in Methods in Molecular Biology (New York, NY, United States), ed. Z. A. Popper, Springer, 2011, Vol. 715, Plant Cell Wall - Methods and Protocols, p. 179. 

Detergent Methods. The neutral detergent fiber (NDF) and acid detergent fiber (ADF) methods (2), later modified for human foods (13), measure total insoluble plant cell wall material (NDF) and the cellulose—lignin complex (ADF). The easily solubilized pectins and some associated polysaccharides, galactomaimans of legume seeds, various plant gums, and seaweed polysaccharides are extracted away from the NDF. They caimot be recovered easily from the extract, and therefore the soluble fiber fraction is lost. 

For monitoring the extent of polysaccharide hydrolysis, l.c. methods that sepeu ate and analyze the non-fermentable oligosaccharides (d.p. 3-30) derived from cellulose, hemicellulose, and pectins are useful, and have already been described (see Section III,l,c). For determination of the monosaccharide composition of completely hydrolyzed, plant polysaccharides, l.c. is especially useful and has been applied to the compositional analysis of hydrolyzed plant fiber,wood pulps,plant cell-walls,and cotton fibers.In these representative examples, the major sugars of interest, namely, glucose, xylose, galactose, arabinose, and mannose, have traditionally been difficult to resolve by l.c. The separa-... 

There are many modifications of this method, they may be to make the process more representative of the ruminant digestive system, or the desired residue may be just the plant cell walls. It is not always possible to say that one procedure is better than another, therefore the chosen procedure may be that which has been used by workers involved in animal nutrition over a number of years in a certain geographical area. The decision may be to use the usual procedure favoured by the referees for research papers in a particular journal. 

Method 7.5. Determination of neutrai detergent fibre (NDF) or plant cell-wall constituents... 

This is based on the method by Van Soest and Wine (1967) which has been modified according to subsequent recommendations. It is the only fibre determination suitable for non-ruminants. The residue consists of the plant cell-wall constituents cellulose, hemicellulose, lignin, cutin, NDF-insoluble tannin and ash. See the article by Cherney (2000) for current modifications these include the use of amylase to aid in the removal of starch from forages containing grain (Van Soest et al., 1991), which has been adopted by MAFF... 

Polne-Fuller, M. (1988) Microorganisms and methods fca- d ading plant cell walls and conplex hydrocarbons. Eur. Pat. Appl. EP 289,908 Chon. Abstr., 1989,110, 151224s. 

Findings on the antiviral activity of 2 -C-methyl- and 3 -C-methyl-cy-tidines, and on the presence of poly-I in plant cell-walls have stimulated the synthesis of branched-sugar nucleosides90 by the usual methods, and some of their physiological activities have been summarized.405... 

A further problem is that many of the methods for plant cell-wall preparation, based upon insolubility in aqueous buffers, salt solutions, and organic solvents, undoubtedly remove, or detach, some of the molecules, present in the intact wall, which may have important, structural functions. 

Selvendran, R.R. and Ryden, P. 1990. Isolation and analysis of plant cell walls. In Methods in Plant Biochemistry, Vol. 2, Carbohydrates (P.M. Dey ed.) pp. 549-579. Academic Press, London. 

Albersheim, P Nevins, D.J., English, P.D., and Karr, A. 1967. A method for the analysis of sugars in plant cell-wall polysaccharides by gas-liquid chromatography. Carbohvdr. Res. 5 340-345. 

Amino sugar derivatives, Morgan-Elson detection method, 656-659 a-Amylase degradation, 716 in plant cell wall isolation, 706 in total starch determination, 685... 

The standard procedure by Saeman et al. (I) involves manual stirring of the polysaccharide with 72% H2S04, standing at 30°C, and secondary hydrolysis at 100° or 120°C in a steam autoclave. While certain resistant polysaccharides are still incompletely depolymerized, decomposition of the more sensitive monosaccharides formed cannot be avoided. An alternative method by using trifluoroacetic acid was applied successfully for plant cell wall polysaccharides by Albersheim et al. (2) and for dissolving pulps and hemicelluloses by Fengel et al. (3). Highly crystalline cellulose was not well dissolved and not completely hydrolyzed by CFsCOOH. 

Classic methods in use to fractionate plant cell wall polysaccharides are mainly based upon the differential solubilities of various cell wall polysaccharide constituents (32). To solubilize pectic polysaccharides, the cell wall material is extracted with water, ammonium oxalate or disodium EDTA. The solubilization of pectic polysaccharides could then be followed by solubilization of hemicelluloses, i.e., polysaccharides rich in D-glucuronic acid 4-0-methyl-D-glucuronic acid, D-xylose and D-glucose residues. The solubilization of hemicellulosic polysaccharides is accomplished by treatment of water, ammonium oxalate or EDTA insoluble material with a strong alkali. 

The wet chemical and rapid analytical methods described above are being used on a daily basis in the U.S. Department of Energy s Office of the Biomass Program. The methods are used primarily to keep track of chemical fractions of plant cell walls... 

The composition of lignin (an important component of plant cell walls) is very complex.p-Coumaric acid, cinnamic acid, ferulic acid, and others are among the products of lignin biodegradation (Fig. 1). Thin layer chromatography is a very rapid method for their separation and is usually completed in a very short time. Six different solvents were used as developer (Table 5), with silica gel being used as a stationary phase. 

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