Inositol Metabolism in Plants. III. Conversion of Myo-inositol-2-H to Cell Wall Polysaccharides in Sycamore (Acer pseudoplatanus L.) Cell Culture

Prolonged growth of cell cultures of sycamore (Acer pseudoplatanus L.) on agar medium containing myo-inositol-2-(3)H resulted in incorporation of label predominately into uronosyl and pentosyl units of cell wall polysaccharides. Procedures normally used to distinguish between pectic substance and hemicellulose yielded carbohydrate-rich fractions with solubility characteristics ranging from pectic substance to hemicellulose yet the uronic acid and pentose composition of these fractions was decidedly pectic. Galacturonic acid was the only uronic acid present in each fraction. Subfractionation of alkali-soluble (hemicellulosic) polysaccharide by neutralization followed by ethanol precipitation gave 3 fractions, a water-insoluble, an ethanol-insoluble, and an ethanol-soluble fraction, each progressively poorer in galacturonic acid units and progressively richer in arabinose units; all relatively poor in xylose units.Apparently, processes involved in biosynthesis of primary cell wall continued to produce pectic substance during cell enlargement while processes leading to biosynthesis of typically secondary cell wall polysaccharide such as 4-0-methyl glucuronoxylan were not activated.     

Sugar composition of pollen grain and pollen tube cell walls

The sugar composition of pollen grain and pollen tube cell walls was studied for Camellia japonica, C. sasanqua, C. sinensis, Tulipa gesneriana and Lilium longiflorum. In all species, the main components of pollen grain walls were arabinose, galactose, glucose and uronic acid. On the other hand, the pollen tube walls consisted mostly of glucose. The pollen tube wall of C. japonica was fractionated into hemicellulose, α-cellulose and pectic substance fractions in yields of 61, 19 and 3 %, respectively. The hemicellulose fraction was composed essentially of glucose. The sugar composition of the pollen tube wall was not influenced by the nature of exogenously supplied sugars. Rapid growth of the pollen tube seemed to correlate with the synthesis of hemicellulosic glucan.     

Turnover of cell wall polysaccharides of a Vinca rosea suspension culture

Turnover of cell wall components was examined in two growth phases of a batch suspension culture of Vinca rosea L. Three-day-cultured cells (cell division phase) and 5-day-cultured cells (cell expansion phase) were incubated with d -[U-¹⁴C]glucose. After various periods of incubation, extra-cellular polysaccharides (ECP) and cell walls were isolated, and then the cell walls were fractionated to pectic substance, hemicellulose, and cellulose fractions. The results of the measurement of radioactivities and amounts of total carbohydrate in the ECP and cell wall fractions indicated that synthesis of pectic substance was more active in the cell division phase than in the cell expansion phase. From the results of the pulse-chase experiments, in which cells prelabelled by incubation with d -[U-¹⁴C]glucose for 3 h were incubated in a medium containing unlabelled glucose for various periods, the gross degradation, net synthesis, and gross synthesis of cell wall components were estimated. Active degradation and synthesis were observed in the hemicellulose fraction, indicating that active turnover occurred in the hemicellulose fraction, while little degradation was found in the pectic substance and cellulose fractions.     

Turnover of cell wall polysaccharides of a Vinca rosea suspension culture

Three-day-cultured cells of Vinca rosea L. (in the cell division phase) and 5-day-cultured cells (in the cell expansion phase) prelabelled with d -[U-¹⁴C] glucose were incubated in a medium containing unlabelled glucose. After various periods of chase, extra-cellular polysaccharides (ECP) and cell walls were isolated, and cell walls were fractionated into pectic substances, hemicellulose, and cellulose fractions. After acid hydrolysis, the radioactive constituents in the pectic substances and hemicellulose fractions were analyzed. Active turnover was observed in arabinose and galactose in the hemicellulose fraction of cell walls, while the constituents of the pectic substances, and xylose and glucose in the hemicellulose fraction did not undergo active turnover. The proportion of radioactivities of arabinose and galactose in total radioactivity of ECP increased markedly after chasing. These results indicate that arabinogalactan was synthesized, deposited in the cell wall, degraded rapidly, and made soluble in the medium as a part of ECP.     

Cell Wall Metabolism in Ripening Fruit: I. CELL WALL CHANGES IN RIPENING ;BARTLETT' PEARS

`Bartlett' pear (Pyrus communis) fruits were picked at the mature, green stage and ripened at 20 C. Fruits at different stages of ripeness (based on flesh firmness) were homogenized, and the sugar and uronic acid contents of cell wall and soluble polysaccharides were determined. Substantial amounts of galacturonic acid and arabinose were lost from the wall fraction as the fruit ripened. Most of this cell wall material was recovered, in an 80% (volume/volume) ethanol-insoluble form, from the soluble fraction of tissue homogenates. Structural analysis of ethanol-precipitable material indicates that it is an acidic (pectic) polymer-bearing side groups containing variously-linked arabinosyl residues.     

Studies on the Pectic Substances of Plant Cell Walls: III. DEGRADATION OF CARROT ROOT CELL WALLS BY ENDOPECTATE LYASE PURIFIED FROM ERWINIA AROIDEAE

Pectate lyase was isolated from the cell extract of Erwinia aroideae. The enzyme was further purified to a high degree by a procedure involving ammonium sulfate fractionation and chromatography on CM-Sephadex C-50 and on Sephadex G-200. The enzyme attacked its substrate in an endo fashion and was more active on the sodium salt of acid-insoluble polygalacturonate or pectic acid than it was on the methoxylated pectin. The enzyme had an optimum pH at 9.3, was stimulated by calcium ions, and was completely inhibited by ethylenediaminetetraacetic acid. In addition, the reaction products showed an absorption maximum between 230 and 235 nm and reacted with thiobarbituric acid. These results indicate that the purified enzyme is an endopectate lyase. The endopectate lyase also had the ability to solubilize effectively the pectic fraction from the cell walls of carrot (Daucus carota) root tissue. The enzyme released 30.5% of the wall as soluble products and also liberated all of the galacturonic acid present in the walls. The total neutral sugars released by the enzyme were 10.6% of the walls, which corresponded to 71.5% of noncellulosic neutral sugars. The soluble products were separated into five fractions by DEAE-Sephadex A-50 column chromatography. Based on the analysis of sugar composition of each fraction, the pectic fraction of carrot cell wall is presented.     

Structure of the Primary Cell Walls of Suspension-Cultured Rosa glauca Cells: I. Polysaccharides Associated with Cellulose

Cell walls of suspension-cultured cells of Rosa glauca were fractionated by two different extraction procedures. The first involved a stepwise fractionation scheme based on alkaline extraction. The second took advantage of the powerful cellulose solvent system N-methylmorpholine N-oxide/dimethyl sulfoxide which is capable of solubilizing whole cell walls. From the analytical composition of each solubilized fraction and of the corresponding residues, the fate of each type of cell wall polysaccharide constituent was followed at each step of the extraction scheme and the mode of action of the extractant was interpreted. Although the two fractionation procedures were very different, they yielded very similar cellulosic complex residues and extracts, thus delimiting two blocks of polysaccharides in the cell wall. The cellulose residues still comprised uronic acid-containing polysaccharides and hemicelluloses in association with cellulose. Graded acid hydrolysis provided evidence for the central role of a homogalacturonan core interconnecting xyloglucans and arabinogalactans. A tentative model showing the possible interaction existing between the constituent polysaccharides still associated to cellulose after alkaline extraction is presented. Hydrogen bonding between xyloglucan and cellulose is confirmed, and glycosidic linkages between xyloglucans and pectic polymers are suggested.     

Cell Wall Dissolution in Ripening Kiwifruit (Actinidia deliciosa) : Solubilization of the Pectic Polymers

Pectic polysaccharides solubilized in vivo during ripening, were isolated using phenol, acetic acid, and water (PAW) from the outer pericarp of kiwifruit (Actinidia deliciosa [A. Chev.] C.F. Liang and A.R. Ferguson var deliciosa `Hayward') before and after postharvest ethylene treatment. Insoluble polysaccharides of the cell wall materials (CWMs) were solubilized in vitro by chemical extraction with 0.05 molar cyclohexane-trans-1,2-diamine tetraacetate (CDTA), 0.05 molar Na₂CO₃, 6 molar guanidinium thiocyanate, and 4 molar KOH. The Na₂CO₃-soluble fraction decreased by 26%, and the CDTA-soluble fraction increased by 54% 1 day after ethylene treatment. Concomitantly, an increase in the pectic polymer content of the PAW-soluble fraction occurred without loss of galactose from the cell wall. The molecular weight of the PAW-soluble pectic fraction 1 day after ethylene treatment was similar to that of the Na₂CO₃-soluble fraction before ethylene treatment. Four days after ethylene treatment, 60% of cell wall polyuronide was solubilized, and 50% of the galactose was lost from the CWM, but the degree of galactosylation and molecular weight of pectic polymers remaining in the CWMs did not decrease. The exception was the CDTA-soluble fraction which showed an apparent decrease in molecular weight during ripening. Concurrently, the PAW-soluble pectic fraction showed a 20-fold reduction in molecular weight. The results suggest that considerable solubilization of the pectic polymers occurred during ripening without changes to their primary structure or degree of polymerization. Following solubilization, the polymers then became susceptible to depolymerization and degalactosidation. Pectolytic enzymes such as endopolygalacturonase and β-galactosidase were therefore implicated in the degradation of solubilized cell wall pectic polymers but not the initial solubilization of the bulk of the pectic polymers in vivo.     

Changes in Esterification of the Uronic Acid Groups of Cell Wall Polysaccharides during Elongation of Maize Coleoptiles

Cell walls of grasses have two major polysaccharides that contain uronic acids, the hemicellulosic glucuronoarabinoxylans and the galactosyluronic acid-rich pectins. A technique whereby esterified uronic acid carboxyl groups are reduced selectively to yield their respective 6,6-dideuterio neutral sugars was used to determine the extent of esterification and changes in esterification of these two uronic acids during elongation of maize (Zea mays L.) coleoptiles. The glucosyluronic acids of glucuronoarabinoxylans did not appear to be esterified at any time during coleoptile elongation. The galactosyluronic acids of embryonal coleoptiles were about 65% esterified, but this proportion increased to nearly 80% during the rapid elongation phase before returning to about 60% at the end of elongation. Methyl esters accounted for about two-thirds of the total esterified galacturonic acid in cell walls of unexpanded coleoptiles. The proportion of methyl esters decreased throughout elongation and did not account for the increase in the proportion of esterified galactosyluronic acid units during growth. The results indicate that the galactosyluronic acid units of grass pectic polysaccharides may be converted to other kinds of esters or form ester-like chemical interactions during expansion of the cell wall. Accumulation of novel esters or ester-like interactions is coincident with covalent attachment of polymers containing galactosyluronic acid units to the cell wall.     

Molecular size and separability features of pea cell wall polysaccharides : implications for models of primary wall structure

Relative molecular size distributions of pectic and hemicellulosic polysaccharides of pea (Pisum sativum cv Alaska) third internode primary walls were determined by gel filtration chromatography. Pectic polyuronides have a peak molecular mass of about 1100 kilodaltons, relative to dextran standards. This peak may be partly an aggregate of smaller molecular units, because demonstrable aggregation occurred when samples were concentrated by evaporation. About 86% of the neutral sugars (mostly arabinose and galactose) in the pectin cofractionate with polyuronide in gel filtration chromatography and diethylaminoethyl-cellulose chromatography and appear to be attached covalently to polyuronide chains, probably as constituents of rhamnogalacturonans. However, at least 60% of the wall's arabinan/galactan is not linked covalently to the bulk of its rhamnogalacturonan, either glycosidically or by ester links, but occurs in the hemicellulose fraction, accompanied by negligible uronic acid, and has a peak molecular mass of about 1000 kilodaltons. Xyloglucan, the other principal hemicellulosic polymer, has a peak molecular mass of about 30 kilodaltons (with a secondary, usually minor, peak of approximately 300 kilodaltons) and is mostly not linked glycosidically either to pectic polyuronides or to arabinogalactan. The relatively narrow molecular mass distributions of these polymers suggest mechanisms of co- or postsynthetic control of hemicellulose chain length by the cell. Although the macromolecular features of the mentioned polymers individually agree generally with those shown in the widely disseminated sycamore cell primary wall model, the matrix polymers seem to be associated mostly noncovalently rather than in the covalently interlinked meshwork postulated by that model. Xyloglucan and arabinan/galactan may form tightly and more loosely bound layers, respectively, around the cellulose microfibrils, the outer layer interacting with pectic rhamnogalacturonans that occupy interstices between the hemicellulose-coated microfibrils.     

Effects of ABA on Synthesis of Cell-Wall Polysaccharides in Segments of Etiolated Squash Hypocotyl I. Changes in Incorporation of Glucose and myo-Inositol into Cell-Wall Components

Externally supplied [³H]myo-inositol and [¹⁴C]glucose were incorporatedin cell-wall fractions of segments of etiolated squash hypocotyl.The extent of incorporation of [¹⁴C]glucose into cell-wall fractionswas very much greater than that of [³H]myo-inositol. Radioactivityfrom [¹⁴C]-glucose was effectively incorporated into hemicelluloseB and cellulose fractions and was incorporated uniformly intohexose, pentose and uronic acid residues, but radioactivityfrom [³H]myo-inositol was incorporated predominantly into uronicacid and pentose residues in the pectin and hemicellulose Bfractions. Exogenously applied ABA significantly suppressed the elongationof segments of squash hypocotyl and the incorporation of radioactivityfrom [^l4C]glucose and [³H]myo-inositol into the segments. Furthermore,ABA significantly inhibited the distribution of incorporatedradioactivity from [¹⁴C]glucose into the cellulose fraction,but did not affect distribution into the pectic fraction. Bycontrast, ABA only slightly inhibited the distribution of theincorporated radioactivity from [³H]myo-inositol into the pecticfraction. These results suggest that most of the cell-wall polysaccharidesin segments of squash hypocotyl are synthesized via the UDP-sugarpathway, and that ABA significantly inhibits the synthesis ofcellulose but not the synthesis of pectic polysaccharides whenABA suppresses the elongation of the segments. (Received March 25, 1988; Accepted November 15, 1988)     

Polysaccharide and oligosaccharide changes in germinating lupin cotyledons

In germinating lupin cotyledons, there was a rapid depletion of raffinose series oligosaccharides, a temporary increase in sucrose and constant low levels of reducing monosaccharides. The major polysaccharide fraction was extracted with hot NH₄ oxalate—EDTA solution and had the constitution of intercellular/cell wall polysaccharide. GLC examination of component sugars showed that as cotyledons expanded this fraction was depleted and that there was selective hydrolysis of arabinose and galactose, so that the uronic acid proportion increased. Gel and DEAE-cellulose chromatography showed that this fraction became more heterogeneous. The neutral and acidic fractions were separated and the component sugars, viscosities, gel chromatographic behaviour and sedimentation constants of these determined. The results indicated that in the later phase of plant cell wall expansion in germinating lupin cotyledons the arabinogalactan side chains of the pectic polysaccharide fraction are selectively hydrolysed leaving a primary wall with a high uronic acid content.     

The Structure of Plant Cell Walls: IV. A Structural Comparison of the Wall Hemicellulose of Cell Suspension Cultures of Sycamore (Acer PseudoPlatAnus) and of Red Kidney Bean (Phaseolus Vulgaris)

The molecular structure and chemical properties of the hemicellulose present in the isolated cell walls of suspension cultures of sycamore (Acer pseudoplatanus) cells has recently been described by Bauer et al. (Plant Physiol. 51: 174-187). The hemicellulose of the sycamore primary cell wall is a xyloglucan. This polymer functions as an important cross-link in the structure of the cell wall; the xyloglucan is hydrogen-bonded to cellulose and covalently attached to the pectic polymers.     

Changes of Cell Wall Composition and Polymer Size in Primary Roots of Cotton Seedlings Under High Salinity

The aim of this study was to investigate changes in cell wallchemical composition and polymer size in the root tip of intactcotton seedlings (Gossypium hirsutum L. cv. Acala SJ-2) grownin saline environments, in order to relate the interaction betweenhigh salinity and root growth to possible changes in cell wallmetabolism. Cotton seedlings were grown in modified Hoagland nutrient solutionwith various combinations of NaCl and CaCl₂. Cell walls werefractionated into four fractions (pectin, hemicellulose 1 and2, cellulose), and analysed for their total sugar content, neutralsugar composition and size of polysaccharides. At 1 mol m^–3Ca, 150 mol m^–3 NaCl resulted in a significant increasein the cell wall uronic acid content, but a reduction in cellulosecontent on a per unit dry weight basis. Supplemental Ca overcamethe inhibitory effect of high Na on cellulose content. The neutralsugar composition of the cell wall fractions showed no majorchanges caused by varied Na/Ca ratios. Determinations of polysaccharidepolymer size showed that high Na at 1 mol m^–3 Ca led toan increase in the amount of polysaccharides of intermediatemolecular size and a decrease in that of small size in the hemicellulose1 fraction, indicating a possible inhibition of polysaccharidedegradation by high Na. This change was not observed in the10 mol m^–3 Ca treatments. The results reveal a relationshipbetween the effects of high salinity on root growth and cellwall metabolism, particularly in regard to cellulose biosynthesis Key words: Gossypium hirsutum, salinity, root, cell wall     

Fractionation and Partial Characterization of Pectic Polysaccharides in Cell Walls from Liverwort (Marchantia polymorph) Cell Cultures

Konno, H., Yamasalu, Y. and Katoh, K. 1987. Fractionation andpartial characterization of pectic polysaccharides in cell wallsfrom liverwort (Marchantia polymorpha) cell cultures.—Jexp. Bot. 38: 711–722. Pectic polysaccharides were extracted from the starch-free cellwall preparation of cell suspension cultures of Marchantia polymorpha.The polysaccharides were fractionated by DEAE-Sephadex A-50ion-exchange chromatography yielding the five fractions, andthe degree of polymerization and glycosyl composition determinedfor each fraction. The neutral rich and acidic pectic polymerswere depolymerized by purified endoglucanase (l,4-ß-D-glucan4-glucanohydrolase, E.C. 3.2.1.4 [EC] .) and endopolygalacturonase(poly-l,4--Dgalacturonide glycanohydrolase, E.C. 3.2.1.15 [EC] ),respectively. The degraded pectic fractions were fractionatedby gel filtration chromatography on Bio-Gel A-5m and Bio-GelP-2, and glycosyl composition determined for each fraction.The results indicate that pectic polysaccharides contain glucose-richpolymer, rhamnogalacturonan and homogalacturonan in a ratioof 1:4:0–6. In addition, pectic polysaccharides were releasedas five pectic fragments from the cell walls by purified endopectatelyase (poly-l,4--D-galacturonide lyase, E.C. 4.2.2.2 [EC] ). Basedon the analysis of glycosyl composition of each fragment, thepectic polysaccharides of Marchantia cell walls are characterized Key words: Cell suspension culture, cell wall, liverwort, Marchantia polymorpha, pectic polysaccharides     

Effects of gibberellic Acid, calcium, kinetic, and ethylene on growth and cell wall composition of pea epicotyls

The influence of gibberellic acid (GA), calcium, kinetin, and ethylene on growth and cell-wall composition of decapitated pea epicotyls (Pisum sativum L. var. Alaska) was investigated. Calcium, kinetin, and ethylene each caused an inhibition of GA-induced elongation of pea stems. Gibberellic acid did not reverse the induction of swelling by Ca²⁺, kinetin, or ethylene. Both Ca²⁺ and ethylene significantly inhibited the stimulatory effects of GA on the formation of residual wall material. Although GA promoted the development of walls relatively low in pectic substances and pectic uronic acid, Ca²⁺, kinetin, and ethylene favored the formation of walls rich in these constituents. Calcium, kinetin, and GA, alone or in combination, had no effect on the production of ethylene by pea epicotyls.     

Enzymic Analysis of Feruloylated Arabinoxylans (Feraxan) Derived from Zea mays Cell Walls : II. Fractionation and Partial Characterization of Feraxan Fragments Dissociated by a Bacillus subtilis Enzyme (Feraxanase)

Structural features of feruloylated arabinoxylan (feraxan) present in Zea mays L. (hybrid B 73 × Mo 17) coleoptile cell walls have been studied using a purified feraxan-dissociating enzyme (feraxanase) and an α-arabinofuranosidase. This experimental approach has demonstrated the following. (a) Feraxanase dissociated ca. 20% (dry weight basis) of the maize wall preparation. The predominant oligosaccharides enzymically liberated were allocated into seven major subfractions designated A-1 (0.8%), B-1 (1.6%), B-2 (2.4%), B-3 (4.6%), C-1 (1.0%), C-2 (4.2%), and C-3 (0.3%). Values in parentheses reflect the percentage of the wall associated with each subfraction. Subfractions represent samples enriched in different degrees of polymerization, sugar composition, linkage arrangements, and phenolic acid content. (b) B-1, B-2, and B-3 fractions are not feruloylated and have smaller molecular mass (less than 10⁴ kilodaltons) and consist chiefly of t-arabinosyl-5-arabinosyl, 4-xylosyl, 2,4/3,4-xylosyl, and glucuronosyl residues, suggesting that these fragments constitute nonferuloylated regions of arabinoxylan. (c) C-2 and C-3 fractions contain ferulic acid (6.2% and 12.1%, respectively) and are similar to the B series in their sugar linkage arrangements but were derived from feruloylated regions. (d) Alkali treatment of the C-2 fraction decreases the molecular size of the fragment and liberates phenolic acids. The results suggest the presence of alkaline-labile links, probably diferulate bridges. (e) A-1 and C-1 fractions are larger (more than 5 × 10⁵ kilodalton) and contain t-galactosyl-, 4-galactosyl, 2,4-rhamnosyl-residues, galacturonic acid, and the sugar linkage arrangements common to other fractions. The A-1 fraction is not feruloylated, whereas C-1 fraction contains 0.5% ferulic acid. The presence of galactose, rhamnose, and galacturonic acid suggests that pectic polymers, probably homopolygalacturonans and rhamnogalacturonans, are linked to nonferuloylated and feruloylated segments of arabinoxylans.     

Effects of calcium and kinetin on growth and cell wall composition of pea epicotyls

Kinetin and CaCl₂, in the presence of indoleacetic acid, promoted lateral expansion of epicotyls of decapitated and derooted Alaska pea seedlings (Pisum sativum L.) and inhibited their elongation. This growth response was correlated with the development of cell walls unusually rich in pectic uronic acids. Epicotyls in calcium-auxin solutions continued to enlarge and to add new wall material long after tissues in auxin only had stopped. Longitudinal enlargement, associated with the development of walls poor in pectic uronic acids, was favored by KCl, MgCl₂, and ethylenediaminetetraacetate. The last of these agents promoted the loss of ⁴⁵Ca from the epicotyls. Seedings grown in vermiculite moistened with CaCl₂, KCl, or MgCl₂ solutions did not differ in appearance or in the composition of their walls. They responded similarly to experimental treatment except that the decapitated epicotyls of the MgCl₂-grown plants suffered an absolute loss of pectic uronate when incubated in that salt.     

Effects of fungus inoculation and salt stress on physiology and biochemistry of in vitro grapevines: Emphasis on sugar composition changes by FT-IR analyses

The impacts of salt stress and inoculation in in vitro grapevine (Vitis vinifera L.) growth, nutrient accumulation, osmoregulation, photosynthesis and membrane integrity were evaluated. One month exposure to 100 mM NaCl as well as to inoculation with Phaeomoniella chlamydospora reduced relative growth rate (RGR) and induced senescence in grapevine plants, shown by: (1) decrease of Ψ_π without osmoregulation, (2) decrease of chlorophyll content and fluorescence, (3) loss of membrane integrity and (4) nutritional disorders. To assess putative changes in structural and/or non-structural carbohydrates induced by these two stress conditions, alcohol insoluble residues from the roots, stems and leaves were also characterised by FT-IR and GC with respect to the sugar composition. The referred organs were distinguished based on: (1) higher proportion of uronic acid residues in leaves which diagnose the presence of pectic polysaccharides (wavenumbers 1100, 1150 and 1018 cm^?1 in FT-IR spectra), (2) higher proportion of xylose and glucose on stems and FT-IR spectra diagnostic of xylose-rich polysaccharides (1041 cm^?1) and cellulose (1060 cm^?1), (3) higher proportion of glucose residues, xylose and arabinose on roots and a FT-IR spectra characteristic of xylose-rich polysaccharides (1041 cm^?1). The main alterations induced by salt stress and inoculation were more visible in leaves, where the content of uronic acid decreased showing that changes in cell wall composition occurred, mostly at the pectic fraction. Besides, an accumulation of insoluble glucose was found, and FT-IR spectra showed that this glucose-based material was starch (maximum absorption at 998 cm^?1), accumulated as a non-specific response to salt stress and P. chlamydospora inoculation.     

Pectic polysaccharide breakdown of cell walls in cucumber roots grown with calcium starvation

Pectic polysaccharides from the roots of cucumber (Cucumis sativus L.) grown in liquid culture medium with or without calcium (1 mm CaCl₂) were studied after extraction successively by hot water and Na hexametaphosphate solution. The Ca²⁺ starvation-treatment caused a striking reduction in content of extracted pectic polysaccharide; from an equivalent weight of cell walls, only 33.1% of the control level was extracted from root cell walls of plants cultured under Ca²⁺ deficiency. The extracted pectic polysaccharides were fractionated into neutral and acidic polymers by a DEAE-Sephadex column. The acidic polymers, which represented more than 76% of the yield, appeared to be a major fraction of extracted pectic polysaccharides. The changes of molecular size and glycosyl residue composition of this fraction were compared for the control and Ca²⁺-deprived samples. The results indicate that Ca²⁺ deficiency caused structural changes which could involve both branching pattern and extent of contiguous galacturonosyl units in the water-solubilized pectic polysaccharides. Ca²⁺ starvation also led to a notable decrease in molecular size of the hexametaphosphate-solubilized polysaccharides and, to a lesser extent, of the water-solubilized fraction as well. In addition, polygalacturonase activity in tissue homogenates increased remarkably with the Ca²⁺ deficiency, whereas β-galactosidase activity did not undergo a change. Thus, it appears that one major effect of Ca²⁺ deprivation was to stimulate polygalacturonase activity, an effect which could be involved in the control of the breakdown of pectic polysaccharides in the cell walls.