Biosynthesis of Xyloglucan in Suspension-cultured Soybean Cells. Evidence that the Enzyme System of Xyloglucan Synthesis does not Contain {beta}-1,4-Glucan 4-{beta}-D-Glucosyltransferase Activity (EC 2.4.1.12)

Xyloglucan 4-ß-D-glucosyltransferase, an enzyme responsiblefor the formation of the xyloglucan backbone, in a particulatepreparation of soybean cells has been compared with ß-1,4-glucan4-ß-D-glucosyltransferase of the same origin. Thefollowing observations indicate that the enzyme system of xyloglucansynthesis does not contain ß-1,4-glucan 4-ß-D-glucosyltransferaseactivity, although both enzymes transfer the glucosyl residuefrom UDP-glucose to form the ß-1,4-glucosidic linkage:1. The incorporation of [¹⁴C]glucose into xyloglucan dependedon the presence of UDP-xylose in the incubation mixture. 2.No measurable amount of radioactivity was incorporated fromUDP-[¹⁴C]xylose into the cello-oligosaccharides, although theincorporation of [¹⁴C]xylose into xyloglucan depended on thepresence of UDP-glucose in the incubation mixture (Hayashi andMatsuda 1981b). 3. The activity of xyloglucan 4-ß-D-glucosyltransferasewas stimulated more strongly by Mn²⁺ than by Mg²⁺, whereas Mg²⁺was the most active stimulator for the activity of ß-1,4-glucan4-ß-D-glucosyltransferase. 4. An addition of GDP-glucose(100 µM) to the incubation mixture inhibited the activityof xyloglucan 4-ß-D-glucosyltransferase by 17%, whereasthe activity of ß-1,4-glucan 4-ß-D-glucosyltransferasewas inhibited 56% under the same conditions. 5. Irpex exo-cellulasedid not hydrolyze the xyloglucan synthesized in vitro. 6. Theß-1,4-glucan synthesized in vitro was not a branchedxyloglucan because it gave no 2,3-di-O-methyl glucose derivativeon methylation analysis. 7. Pulse-chase experiments indicatedthat the ß-1,4-glucan was not transformed into thexyloglucan. The subcellular distribution of the xyloglucan synthase, however,was similar to that of the ß-1,4-glucan synthase (Golgi-located1,4-ß-D-glucan 4-ß-D-glucosyltransferase).Thus, it appears that the latter enzyme is located at a siteclose to xyloglucan synthase and is set aside for the assemblyof these polysaccharides into the plant cell surface. (Received May 21, 1981; Accepted October 13, 1981)     

Enzymic Dissociation of Zea Shoot Cell-Wall Polysaccharides V. Dissociation of Xyloglucan by Urea

A procedure has been devised to extract and identify structuralcomponents of the xyloglucan of Zea mays L. (hybrid B73 ? Mo17) shoot cell-walls. A water-insoluble fraction of Zea shootcell-walls, after pretreatment with purified Bacillus subtilis(1 3),(1 4)-ß-D-glucan 4-glucanohydrolase, purifiedB. subtilis endo-(l 4)-ß-xylanase and an enzyme preparationfrom B. subtilis enriched in glucuronoxylanase (Kato and Nevins1984a, Nishitani and Nevins 1991), was subsequently treatedwith 7 M urea. The carbohydrates (0.8% of the water-insolublefraction of Zea shoot cell-walls) liberated by the urea treatment,were comprised of xyloglucan polymers with molecular weightswhich varied from 1.0 ? 10⁴ to 4.0 7times; 10⁴ Da. Other wallfragments associated with the isolated polymer suggest covalentbonding of xyloglucan to other polysaccharides. Structural analysesof the xyloglucan polymers reveal a cellulose-like backbonewith about 35% of the C-6 positions substituted with xyloseand other sugars. About 80% of xyloglucan present in the enzyme-pretreatedwater-insoluble fraction of Zea shoot cell-walls was liberatedby the urea treatment. The procedure avoids the use of alkaliin the solubilization of xyloglucan. ¹Supported in part by National Science Foundation research grantsPCM 7818588 and DMB 8505901. (Received September 10, 1990; Accepted May 15, 1991)     

Xyloglucan and r{beta}-D-Glucan in Cell Walls of Rice Seedlings

Cell walls of 4-day old rice seedlings were extracted successivelywith ammonium oxalate-oxalic acid, 4% KOH and 24% KOH. A rß-D-glucanpreparation and a xyloglucan preparation were isolated fromthe 4% KOH extract and 24% KOH extract, respectively. Methylationanalysis and enzymic degradation studies of the polysaccharidesshowed that the former was built up predominantly of repeating-oligosaccharideunits of 3-O-rß-cellobiosyl-D-glucose and 3-O-rß-cellotriosyl-D-glucosein a molar ratio of 2.6 : 1.0, and the latter was of repeating-oligosaccharideunits of -D-xylosyl-(16)-rß-D-glucosyl-(14)-[-D-xylosyl-(16)]-rß-D-glucosyl-(14)-D-glucose,-D-xylosyl-(16)-rß-D-glucosyl-(14)-D-glucose and cellobiose. ¹ Present address: Department of Botany, Iowa State University,Ames, Iowa 50011, U.S.A. (Received August 29, 1981; Accepted January 12, 1982)     

Purification and Properties of an Extracellular Endo-1,4-r{beta}-Glucanase from Suspension-Cultured Poplar Cells

An endo-1,4-rß-glucanase (EC 3.2.1.4 [EC] ) was purifiedto apparent homogeneity from the culture medium of poplar (Populusalba L.) cells by sequential anion-exchange, hydrophobic, andgel-filtration chromatography. The preparation of extracellularrß-glucanase was homogeneous on SDS-polyacrylamidegel electrophoresis (PAGE) and native PAGE. The molecular weight,as determined by SDS-PAGE was 50,000, whereas that determinedby gel filtration was 40,000. The isoelectric point (pI) was5.5. The purified enzyme catalyzed the endohydrolysis of carboxy-methylcellulosewith a pH optimum of 6.0 and a k_m of 1.0 mg ml^–1. Theenzyme specifically cleaved the 1,4-rß-glucosyl linkagesof carboxymethylcellulose, swollen cellulose, lichenan and xyloglucan,although the last was hydrolyzed more slowly than the othertested substrates. The activity of the endo-1,4-rß-glucanaseincreased up to the early stage of the mid-logarithmic phaseof growth and then decreased rapidly, suggesting that the rß-glucanaseis induced before cell development. (Received April 28, 1993; Accepted July 19, 1993)     

Purification and Properties of a Wall-Bound Endo-1,4-{beta}-Glucanase from Suspension-Cultured Poplar Cells

A wall-bound endo-1,4-ß-glucanase (EC 3.2.1.4 [EC] ) wasobtained from a preparation of the cell walls of suspension-culturedpoplar cells and purified to electrophoretic homogeneity bycation-exchange, hydrophobic, and gel-filtration chromatography.The molecular mass was estimated to be 47 kDa by SDS-PAGE and48 kDa by gel filtration on Superdex 200 pg. The isoelectricpoint (pI) was 5.6. The purified enzyme catalyzed the endo-hydrolysisof carboxymethylcellulose with an optimal pH of 6.5, a K_m of1.2 mg ml^-1, and a V_max of 280 units. The purified enzyme specificallyhydrolyzed the 1,4-ß-glucosyl linkages of carboxymethylcellulose,phospho-swollen cellulose, lichenan, xylan and xyloglucan. Theactivity of the enzyme was strongly stimulated by cysteine-HCl.The N-terminal sequence of the enzyme was similar to that ofan extracellular endo-1,4-ß-glucanase found in suspensioncultures of poplar cells and some homology was recognized toavocado fruit-ripening and bean abscission endo-1,4-ß-glucanases. ¹This work was supported in part by a grant from the Toray ScienceFoundation, Japan, and by a Grant-in-Aid from the Ministry ofEducation, Science and Culture of Japan.     

Characterization of the adsorption of Xyloglucan to Cellulose

The binding of xyloglucan- and cello-oligosaccharides to cellulosescan be expressed by Langmuir adsorption isotherms, in whichthe levels of adsorption maxima are all similar but very low.In the present study, although the adsorption constant increasedwith increases in the degree of polymerization (DP) of the 1,4-rß-glucosylresidues of xyloglucan- and cello-oligosaccharides, the adsorptionconstant of cellopentitol to cellulose was similar to that ofhendecosanosaccharide (glucose/xylose, 12 : 9), demonstratingless extensive binding in the case of xyloglucan oligosaccharidesin spite of longer chains of 1,4-rß-glucosyl residues.The binding to cellulose of xyloglucans from pea and Tamarindusindica can also be expressed as Langmuir adsorption isotherms.The adsorption constant for pea xyloglucan with a DP for 1,4-rß-glucosylresidues of 150 was obviously higher than that for Tamarindusxyloglucan with a DP of 3,000. The adsorption maximum and adsorptionconstant of Tamarindus xyloglucan decreased gradually as theDP of 1,4-rß-glucosyl residues decreased from 3,000to 64. This result demonstrates that fucosylated pea xyloglucanhas a higher adsorption constant for cellulose than non-fucosylatedTamarindus xyloglucan when the DP of 1,4-rß-glucosylresidues is identical. These findings indicate that xyloglucanbinds to cellulose as a mono-layer and fucosyl residues contributeto the increase in adsorption affinity. (Received June 4, 1994; Accepted September 10, 1994)     

Xyloglucan in the Cell Walls of Suspension-Cultured Rice Cells

The xyloglucan present in the 24% KOH extract of the cell wallsof suspension-cultured rice cells was characterized by fragmentationanalysis with Trichoderma viride cellulase and Aspergillus oryzaeß-D-glucosidase. The xyloglucan is composed mainlyof the following oligosaccharide units: Results showed that the xyloglucan of suspension-cultured ricecells is more extensively branched than is that of rice seedlings.Another structural characteristic of the former xyloglucan isthe presence of D-galactosyl-D-xylosyl side chains that arenot found in the latter. (Received June 15, 1984; Accepted January 11, 1985)     

Pea Xyloglucan and Cellulose: VI. Xyloglucan-Cellulose Interactions in Vitro and in Vivo

Since xyloglucan is believed to bind to cellulose microfibrils in the primary cell walls of higher plants and, when isolated from the walls, can also bind to cellulose in vitro, the binding mechanism of xyloglucan to cellulose was further investigated using radioiodinated pea xyloglucan. A time course for the binding showed that the radioiodinated xyloglucan continued to be bound for at least 4 hours at 40°C. Binding was inhibited above pH 6. Binding capacity was shown to vary for celluloses of different origin and was directly related to the relative surface area of the microfibrils. The binding of xyloglucan to cellulose was very specific and was not affected by the presence of a 10-fold excess of (1→2)-β-glucan, (1→3)-β-glucan, (1→6)-β-glucan, (1→3, 1→4)-β-glucan, arabinogalactan, or pectin. When xyloglucan (0.1%) was added to a cellulose-forming culture of Acetobacter xylinum, cellulose ribbon structure was partially disrupted indicating an association of xyloglucan with cellulose at the time of synthesis. Such a result suggests that the small size of primary wall microfibrils in higher plants may well be due to the binding of xyloglucan to cellulose during synthesis which prevents fasciation of small fibrils into larger bundles. Fluorescent xyloglucan was used to stain pea cell wall ghosts prepared to contain only the native xyloglucan:cellulose network or only cellulose. Ghosts containing only cellulose showed strong fluorescence when prepared before or after elongation; as predicted, the presence of native xyloglucan in the ghosts repressed binding of added fluorescent xyloglucan. Such ghosts, prepared after elongation when the ratio of native xyloglucan:cellulose is substantially reduced, still showed only faint fluorescence, indicating that microfibrils continue to be coated with xyloglucan throughout the growth period.     

Effects of the Degree of Polymerization on the Binding of Xyloglucans to Cellulose

Xyloglucan oligosaccharides were isolated with various degreesof polymerization (DP) and reduced with tritiated sodium borohydride.The ³H-oligosaccharides were tested for their ability to bindto amorphous and microcrystalline celluloses and to cellulosefilter paper. The time course of binding indicated that theradiolabeled oligosaccharides continued to be bound for at least1 h after heating at 120°C. The binding probably requiredthe organization of the oligosaccharides and celluloses by gradualannealing after heating. Although neither pentasaccharide (glucose:xylose, 3 : 2), heptasaccharide (glucose: xylose, 4 : 3) andnonasaccharide (glucose : xylose : galactose : fucose, 4 : 3: 1 : 1) failed to bind to the celluloses, binding occurredwith oligosaccharides with DP equivalent to more than four consecutive1,4-ß-glucosyl residues. The extent of binding tothe celluloses increased gradually from octasaccharide (glucose:xylose, 5 : 3) to hendecosanosaccharide (glucose/xylose, 12: 9), with the increase in the DP of 1,4-ß-glucosylresidues. The binding of reduced cello-dextrins to celluloserequired at least 4 consecutive 1,4-ß-glucosyl residues.The extent of binding of cellopentitol or cellohexitol to cellulosewas similar to that of hendecosanosaccharide, showing lowerbinding for xyloglucan oligosaccharides in spite of longer chainsof 1,4-ß-glucosyl residues. These findings suggestthat the mode of binding to cellulose of xyloglucan oligosaccharidesis different from that of cello-oligosaccharides. (Received February 18, 1994; Accepted June 1, 1994)     

Endo-1,4-{beta}-Glucanase in the Cell Wall of Stems of Auxin-Treated Pea Seedlings

Endo-1,4-ß-glucanase induced by treatment of pea seedlingswith 2,4-D was extracted from a preparation of the walls ofepicotyl cells. The ß-glucanase was purified by chromatographyon DEAE-cellulose, affinity chromatography on Con A-Sepharoseand SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The activityof ß-glucanase was retained after removal of SDS andextraction from polyacrylamide gels. The band of a protein (46kDa), that corresponded to the activity of endo-1,4-ß-glucanase,was injected directly into mice for preparation of antiserumand the protein was also subjected to amino acid sequencingafter blotting onto a membrane. Western blot analysis showedthat the antiserum obtained bound to a 46-kDa polypeptide andrecognized endo-1,4-ß-glucanase. The N-terminal sequenceof the 46-kDa polypeptide revealed some homology to abscissionendo-1,4-ß-glucanases of bean and avocado fruit. (Received September 29, 1993; Accepted January 20, 1994)     

Cellulase, Fruit Softening and Abscission in Red RaspberryRubus idaeusL. cv Glen Clova

The ripening of raspberry fruit (Rubus ideausL. cv Glen Clova)is associated with a climacteric rise in ethylene production.As the fruit pigments change from green to red there is a progressivesoftening, loss of skin strength and a breakdown of cell wallsin the mesocarp. An increase in cellulase (endo-1,4-ß-D-glucanase)in both drupelets and receptacles accompanies these changes.The localization of cellulase in the regions of the fruit associatedwith abscission zones suggest the enzyme may be involved infruit separation as well as softening. Rubus idaeusL; raspberry; fruit ripening; ethylene; abscission; cell wall breakdown; cellulase; endo-1,4-ß-D-glucanase     

Asparagine biosynthesis in soybean nodules

Two auxin-induced endo-1,4-β-glucanases (EC 3.2.1.4) were purified from pea (Pisum sativum L. var. Alaska) epicotyls and used to degrade purified pea xyloglucan. Hydrolysis yielded nonasaccharide (glucose/xylose/galactose/fucose, 4:3:1:1) and heptasaccharide (glucose/xylose, 4:3) as the products. The progress of hydrolysis, as monitored viscometrically (with amyloid xyloglucan) and by determination of residual xyloglucan-iodine complex (pea) confirmed that both pea glucanases acted as endohydrolases versus xyloglucan. K_m values for amyloid and pea xyloglucans were approximately the same as those for cellulose derivatives, but V_max values were lower for the xyloglucans. Auxin treatment of epicotyls in vivo resulted in increases in net deposits of xyloglucan and cellulose in spite of a great increase (induction) of endogenous 1,4-β-glucanase activity. However, the average degree of polymerization of the resulting xyloglucan was much lower than in controls, and the amount of soluble xyloglucan increased. When macromolecular complexes of xyloglucan and cellulose (cell wall ghosts) were treated in vitro with pea 1,4-β-glucanase, the xyloglucan component was preferentially hydrolyzed and solubilized. It is concluded that xyloglucan is the main cell wall substrate for pea endo-1,4-β-glucanase in growing tissue.     

Purification and Characterization of Wall-bound Exo-l,3-{beta}-D-Glucanase from Barley (Hordeum vulgare L.) Seedlings

A ß-D-glucanase activity hydrolyzing 1,3:1,4-ß-D-glucanwas released from the cell walls of barley by 3M LiCl treatment.It was purified by sequential cation-exchange, gel-filtrationand hydrophobic chromatography. The molecular mass of the glucanasewas 66 kDa as determined by SDS-polyacrylamide gel electrophoresis.Sequence determination of the first thirty amino acids of theN-terminus revealed a high homology of this enzyme to the Pseudomonasl,4-ß-D-glucosidase (56.5%). The purified ß-D-glucanasehas a pH optimum at 5.0, and hydrolyzes oligosaccharides containingß-D-1,3 or ß-D-1,4 linkage. The glucanaseshowed maximum hydrolytic activity toward laminaritetraose,the rate being about two times that of cellotetraose and aboutfour times that of gentiobiose. Polysaccharides such as lichenan,l,3:l,4-ß-D-glucan (from barley), laminarin and pustulanare also hydrolyzed, but not carboxylmethyl-curdlan, carboxymethyl-cellulose,xyloglucan and maltose. The purified ß-D-glucanaseyielded monomeric glucose from laminarihexaose, and exhibitedcharacteristics of an exo-l,3-ß-D-glucanase (EC 3.2.1.58 [EC] ).The activity and biochemical characteristics of this enzymesuggest that it is an exo-l,3-ß-D-glucanase involvedin the rapid turnover of l,3:l,4-ß-D-glucan in barleycell walls during seedling growth. (Received September 24, 1996; Accepted December 9, 1996)     

Glucans in the Cell Walls Regenerated from Vinca rosea Protoplasts

Protoplasts prepared from suspension-cultured Vinca rosea cellswere cultured for 5 days. The cell walls regenerated from theprotoplasts were mainly composed of glucans having 1,3- and1,4-linkages. To investigate the molecular species, these glucanswere separated into four fractions: EDTA (50 mM, pH 4.5)-soluble(fraction E), KOH (24%)- soluble but not precipitatable by neutralizationwith acetic acid (fraction K-S), KOH (24%)-soluble and precipitatableby neutralization with acetic acid (fraction K-P), and KOH (24%)-insoluble(fraction C). By means of sugar composition analysis, methylationanalysis, periodate oxidation and enzymatic digestion, the molecularspecies of the glucans contained in the regenerated cell wallswere deduced to be ß-1,4-glucan (cellulose) and ß-1,3-glucan.Fraction C was mainly composed of ß-1,4-glucan; ß-1,3-glucanwas mainly recovered in fraction K-P. The ß-l,3-glucanwas soluble in dilute alkali solution, but was only slightlysoluble in water. The ß-1,3-glucan had an essentiallyunbranched structure, and its weight average molecular weightestimated by gel permeation chromatography was 4.5–5.0x 10⁴. ¹ Present address: Division of Environmental Biology, NationalInstitute for Environmental Studies, Yatabe, Tsukuba, Ibaraki305, Japan (Received May 21, 1981; Accepted October 13, 1981)     

Characterization of a Non-abscission Mutant in Lupinus angustifolius L.: Physiological Aspects

A single-gene recessive mutant (Abs^-) of Lupinus angustifoliusL. ‘Danja’ that does not abscise any organs wascompared with its parent during continuous exposure of explantsfrom 14 d old seedlings to 10 µl l^-1ethylene. Both endo-(1,4)-ß- D -glucanase (cellulase) and polygalacturonase(PGA) activities increased significantly and progressively inpetiole-stem abscission zones of the parent before the onsetof abscission, and were reflected in a rapid decline in breakstrengthfrom 300 to 70 g within 32 h. In the mutant there was negligibleincrease in hydrolytic enzyme activity, breakstrength declinedslowly (to 180–200 g by 72 h) and there was no abscission.Isoelectric focusing showed two cellulase isoforms (pI 5.0 andpI 8.5) expressed in abscission zones of the parent; these wereexpressed at much lower levels in the mutant. These data areinterpreted to indicate that expression of at least two formsof cellulase activity is enhanced by ethylene in normal petioleabscission zones of lupin. PGA activity also increased in theabscission zone tissue of the parent but to a lesser extentin that of the mutant. We attribute the Abs^-phenotype to mutationof a gene regulating ethylene-responsive expression of abscission-specifichydrolytic enzymes. Copyright 2001 Annals of Botany Company Lupinus angustifolius, abscission, breakstrength, cellulase, ethylene, legume, lupin, mutant, polygalacturonase     

Biosynthesis of Xyloglucan in Suspension-Cultured Soybean Cells. Synthesis of Xyloglucan from UDP-Glucose and UDP-Xylose in the Cell-Free System

When UDP-[¹⁴C]glucose or UDP-[¹⁴C]xylose was incubated witha particulate fraction from soybean cells, radioactive polymerswere synthesized. On digestion with Aspergillus oryzae enzymes,these polymers gave ¹⁴C-monosaccharides and a ¹⁴C-disaccharidewith chromatographic and electrophoretic mobilities indistinguishablefrom those of authentic isoprimeverose (6-O--D-xylopyranosyl-D-glucopyranose).The disaccharide consisted of xylose and glucose, and the latterwas located at the reducing end. Evidence that the disaccharideis isoprimeverose was provided by methylation analysis. Hydrolysisof the methylated disaccharide yielded 2,3,4-tri-O-methyl-D-xyloseand 2,3,4-tri-O-methyl-D-glucose. Thus, incorporation of radioactivityinto isoprimeverose, the smallest structural unit of xyloglucan,suggests that xyloglucan is synthesized in vitro from UDP-glucoseand UDP-xylose. (Received November 20, 1980; Accepted February 14, 1981)     

Degradation of Xyloglucan by Wall-Bound Enzymes from Soybean Tissue II: Degradation of the Fragment Heptasaccharide from Xyloglucan and the Characteristic Action Pattern of the {alpha}-D-Xylosidase in the Enzyme System

Hydrolysis of the fragment heptasaccharide (glucose : xylose= 4 : 3) from xyloglucan with an enzyme preparation from soybeancell wall produced a penta- and a trisaccharide. The resultsof fragmentation analysis of these oligosaccharides with Aspergillusoryzae ß-D-glucosidase indicate the following structuresfor the penta- and trisaccharide. The detection of these intermediate products suggested thatdegradation of the heptasaccharide took place by sequentialsplitting of the -D-xylosidic and ß-D-glucosidic linkages.A characteristic action pattern of the a-D-xylosidase in theenzyme preparation was found. ¹Present address: Department of Biology, McGill University,Montreal, Canada. ²Present address: Department of Botany, Iowa State University,Ames, Iowa 50010, U.S.A. (Received August 20, 1982; Accepted December 7, 1982)     

Trimming and solubilization of xyloglucan after deposition in the walls of cultured rose cells

A pulse-chase technique involving the in vivo feeding of L-[1-³H]arabinoseto suspension-cultured rose (Rosa) cells at 4 d and 9 d aftersubculture (fast- and slow-growing, respectively) was used tocreate a population of [³H]xyloglucan molecules and to followtheir subsequent fate. The weight-average relative molecu larmass (M_w) of [³H]xyloglucan freshly deposited in the cell wallwas 160 000 and 240 000 in the fast- and slow-growing cells,respectively. The wall-bound [³H]xyloglucan of both culturesunderwent a decrease in M_w of 40 000 during the first 2 d afterthe pulse-labelling. At the same time, 20–30% of the initially-deposited[³H]xyloglucan disappeared from the cell wall, and a similaramount appeared in solution in the culture medium. Its failureto remain bound to the cell wall and its low M_w (39 000) indicatedthat this soluble extracellular ( was derived from partial degradationof segments of wall-bound xyloglucan that were not directlyhydrogen-bonded to microfibrils (‘loose ends’ and‘tethers’). The possible enzymic basis and biologicalroles of the degradation are discussed. Key words: Cell expansion, cell wall, hemicellulose, sloughing, xyloglucan     

Macromolecular Complexes of Xyloglucan and Cellulose Obtained by Annealing

Macromolecular complexes composed of xyloglucan and cellulosewere produced by heating amorphous celluloses with xyloglucanin water at temperatures above 160°C. The extraction ofxyloglucan from the annealed specimens required concentratedalkali which might cause microfibrils to swell. Annealed specimensobtained by heating at 200°C had a somewhat fiber-like appearanceeven though mixtures of amorphous celluloses and xyloglucanwere completely amorphous before annealing. Annealing occursspecifically between amorphous celluloses at high temperatures,where xyloglucan may be entrapped into the bundles of cellulosefibers during fiber formation rather than bound to the surfaceof the fibers. (Received September 3, 1993; Accepted December 1, 1993)