Effect of root exudates on the exopolysaccharide composition and the lipopolysaccharide profile of Azospirillum brasilense Cd under saline stress

The effect of wheat root exudates on the exopolysaccharide (EPS) composition and the lipopolysaccharide (LPS) profile of Azospirillum brasilense Cd under saline stress was studied. EPS of A. brasilense Cd was composed of glucose (47%), mannose (3%), xylose (4%), fucose (28%), rhamnose (6%), arabinose (1%) and galactose (11%). Under saline stress, A. brasilense produced a totally different EPS, composed mainly of galactose. Root exudates induced changes in A. brasilense EPS composition only under normal conditions, consisting of higher amounts of arabinose and xylose compared with EPS of bacteria grown without root exudates. No changes were induced by root exudates when A. brasilense was grown under saline stress. Additionally, root exudates induced changes in the LPS profile, both under normal and stress conditions.     

Detection of root mucilage using an anti-fucose antibody

Plant root mucilage is known to enhance soil quality by contributing towards the soil carbon pool, soil aggregation, detoxification of heavy metal ions and interactions with rhizospheric microflora. Mucilage consists of many monosaccharide units, including fucose which can be used as an indicator for plant root based polysaccharides. This is the first report of an immunological technique developed to use anti-fucose antibodies as markers for probing and localizing fucosyl residues in mucilage polysaccharide and, in turn, for localization of plant root mucilage. Fucose was complexed with bovine serum albumin to raise antibodies against fucose. A fucose-directed antibody was shown to cross-react with root cap mucilages from grasses. This antibody was used to localize root mucilage polysaccharide in maize and wheat root caps using immunogold electron microscopy. Abundant labelling could be localized on the cell wall, and in the intercellular matrix and vesicles of the peripheral root cap cells. Labelling was less intense in cells towards the centre of the root cap tissue. Control experiments confirmed that immunogold localization of fucose was specific and reliable.     

Comparison between maize root cells and their respective regenerating protoplasts: wall polysaccharides

The cell-wall polysaccharides from different parts of maize roots have been analysed. The arabinose, galactose and mannose contents are influenced by cell differentiation, whereas xylose, rhamnose and uronic-acid contents are not. In cap cells, the pectin content is low but rhamnose and fucose are present in larger quantities. The cell-wall polysaccharides from cells of the elongation zone and their respective regenerating protoplasts were also analysed. The walls of the protoplasts contained higher xylose and mannose levels and a much lower level of cellulose than the cells from which they were derived.     

Composition of Actinidia mucilage

Purified mucilages extracted from several plant parts of Actinidia chinensis and from the leaves of nine Actinidia species, were shown to be acidic polysaccharides, containing galactose, arabinose, mannose and glucuronic acid. Fucose and xylose were also present in the mucilages from A. chinensis and in the leaf mucilage of four other species. Partial hydrolysis studies suggested that all the mucilages may belong to the glucuronomannan family of polysaccharides, with a repeating disaccharide core of glucuronosylmannose. Division of the Actinidia genus into subgenera may be possible on the basis of properties and monosaccharide compositions of the mucilages.     

Mechanical properties and polysaccharide nature of the cell walls of maize root

Changes in mechanical properties and chemical nature of the cell walls of the different zones along elongating maize ( Zea mays L. cv. LG 11) roots were analyzed and the following results were obtained. (1) The apical region 2 to 5 mm from the tip of 15 mm long roots showed rapid elongation whereas the region 8–10 mm from the tip showed very little growth. (2) The minimum stress-relaxation time (To) and the mean stress-relaxation rate (R) of the cell wall were small whereas the maximum stress-relaxation time (Tm) was large in the region where cell elongation was optimum. The To and R increased and the Tm decreased gradually towards the base of the root. (3) The amounts of non-cellulosic polysaccharides of the cell wall were highest in the region 1.5–2.5 mm from the tip, decreasing until 5 mm from the tip, and then increasing towards the base. However, the proportion of this fraction in the total cell wall polysaccharides was highest in the extreme tip (cap and meristem, 0–1 mm) and decreased towards the base. (4) Major neutral sugars constituting the non-cellulosic polysaccharides of the cell wall were xylose, arabinose, galactose and glucose, with minor amounts of rhamnosc. mannose and fucose. The 1–15 mm region was on the whole rich in glucose and xylose and contained arabinose to a lesser extent. However, the chemical nature in the apical region, (0–2 mm, was rather special, being rich in galactose and fucose. (5) The cell wall of maize roots contained, as a whole, only little pectic substances but was high in hemicellulose 1 (rich in xylose, arabinose and glucose) and hemicellulose 2 (rich in glucose and xylose). (6) It appeared that in the elongating region (apical 2 to 5 mm) the cell elongation rate (CET) showed a rather good correlation with the parameters of mechanical properties (To, Tm and R) and with neutral sugar compositions in the non-cellulosic polysaccharides.     

Golgi Apparatus Mediated Polysaccharide Secretion by Outer Root Cap Cells of Zea mays. III. Control by Exogenous Sugars

Sugars supplied to germinating seedlings of maize (Zea mays L.) regulate the secretion of polysaccharides by the outer cells of the root cap. The polysaccharide secreted by these cells adheres to the root tip as a droplet and the size of the droplet was used to quantitate polysaccharide secretion. The polysaccharide contains glucose, galacrose, and galacturonic acid residues with smaller quantities of mannose, arabinose, xylose, fucose and rhamnose. These sugars supplied to maize seedlings had marked effects on the rate of polysaccharide secretion by root tips. The effects on secretion were independent of the growth rates of the roots. Glucose, fucose and xylose increased droplet size 1.5–2 fold (as did sucrose, maltose, lacrose, fructose and ribose) whereas galactose, arabinose and galacturonic acid were inhibitory. Mannose increased dropler size 5–7 fold. The marked effect of mannose on polysaccharide secretion was due to an increased rate of secretion combined with a longer phase of extrusion of polysaccharide into the forming droplet. The effect of mannose was partially reversed by inorganic phosphate and other sugars (except for fucose which had no effect or promoted secretion in the presence of mannose). In contrast to sucrose, mannose stimulated secretion in a maize variety having a high sugar endosperm (high endogenous sugar). The results suggest that regulation of secretion by mannose is due to an alteration of normal sugar metabolism; whereas stimulation of secretion by sucrose and other sugars may be due to an increased availability of sugars for metabolism.     

Stimulation of polysaccharide formation by 2,4-dichlorophenoxyacetic acid in callus tissues of Arabidopsis thaliana

A relatively high concentration of 2,4-dichlorophenoxyacetic acid (45 μ M ) in solid culture medium stimulated the formation and secretion of mucilage polysaccharides by callus tissues of Arabidopsis thaliana L. Heynh. (line Estland). The mucilage was composed of at least two polysaccharides as revealed by gel chromatography on Sepharose 4B: the major component (87%) eluted in the void volume (molecular weight 2 × 10⁶ or greater) and the minor component (13%) eluted in the molecular weight range from 2 × 10⁴ to 4 × 10⁵. Both polysaccharide components contained small amounts of uronic acids. The major polysaccharide consisted mostly of galactose (49%), arabinose (28%) and fucose (10%), whereas the minor one consisted of galactose (44%), xylose (18%), arabinose (14%) and rhamnose (14%). One of the components of the secreted mucilage seems to be an arabinogalactan.     

Patterns of polysaccharide biosynthesis in differentiating cells of maize root-tips

1. The patterns of incorporation of radioactivity from d-[6-(3)H]-, d-[1-(14)C]-, d-[U-(14)C]- and d-[6-(14)C]-glucose and [U-(14)C]myoinositol into the neutral sugars and uronic acids of the polysaccharides synthesized in different regions of the root-tip of maize were determined. 2. The root-cap tissue synthesized a slime in which a polysaccharide that contained a high proportion of fucose (32%) and galactose (21%) was found. This polysaccharide is synthesized only by the root-cap cells, and very little polysaccharide containing fucose is synthesized in adjacent tissues. Part of the meristematic tissue of the root is surrounded by the cap cells. A section of the root that contains both these tissues can be analysed, and the polysaccharide synthesized by the meristematic region can be obtained since the contribution of the root-cap cells can be found by the amount of fucose formed. 3. It was shown that there is very little difference in the polysaccharide synthesis of the meristematic region from that of the cells immediately behind it. In the more mature cells, however, the amount of xylose synthesized relative to that of arabinose is increased, and the proportion of xylose and arabinose formed in the matrix polysaccharides is increased whereas that of galactose is decreased. 4. The effect of 2,4-dichlorophenoxyacetic acid (2,4-D) on polysaccharide synthesis was to bring about a decrease in the relative amount of galactose synthesized in the matrix polysaccharides of cells immediately adjacent to the meristematic region and also in the more mature tissue. The growth factor also increased the amount of xylose synthesized relative to that of arabinose in the more mature tissue. These metabolic effects were related to a very obvious change in the morphological appearance of the root-tips. 5. Radioactivity from [U-(14)C]myoinositol was incorporated mainly into xylose, arabinose and galacturonic acid rather than into the hexoses, although small amounts of these sugars were formed.     

Plant and bacterial mucilages of the maize rhizosphere: Comparison of their soil binding properties and histochemistry in a model system

Mucilages from the root tips of axenically-grown maize and from a bacterium (Cytophaga sp.) isolated from the rhizosheaths of field-grown roots, were immobilized by drying onto nylon blotting membrane. The mucilage plaques remained in place through repeated rewettings and histochemical treatments. Staining of the plaques showed that both mucilages included acidic groups, and 1,2 diols (the latter notably fewer in bacterial mucilage). Bacterial mucilage plaques stained strongly for protein, plant mucilage was unstained. Plaques of both mucilages bound soil particles strongly if soil was applied to wet mucilage and then dried. Bound soil was not lost with rewetting. Dry weight and densitometer measurements showed that bacterial mucilage bound about 10% more soil than the same surface area of root-cap mucilage. Pretreatment of plaques with periodate oxidation eliminated most soil binding by root-cap mucilage but this was completely reversible by reduction with borohydride. Soil binding to bacterial mucilage was unaffected by periodate but much diminished by borohydride pretreatment (partially restored by subsequent oxidation). Neither pretreatment with cationic dyes nor preincubation in pectinase, pectin methylesterase or protease affected subsequent soil binding by the mucilage plaques. Pretreatment of root-cap mucilage plaques with lectins specific for component sugars also did not alter soil binding. It is concluded that mucilages of both plant and bacterial origin can contribute to the adhesion and cohesion of maize rhizosheaths, but each by a different mechanism. Binding by root-cap mucilage depends on 1,2 diol groups of component sugars, that of bacterial mucilage does not, and is likely to be protein mediated. ei]Section editor: R O D Dixon     

Studies on the secretion of maize root cap slime: I. Some properties of the secreted polymer

The secreted slime from root cap cells of corn (Zea mays, cv. SX-17) was studied. Production of slime by excised root tips is stimulated by the addition of 40 mM sucrose or fucose and half-strength Hoagland's solution to the incubation medium. Secreted slime was recovered from aqueous solution by precipitation with ethanol. The polymer has a molecular weight greater than 2 × 10⁻⁶ daltons and a density of 1.63 g cm⁻³. Protein is not present in material purified by density gradient centrifugation with cesium chloride. Fucose (39%) and galactose (30%) are the principle neutral sugars found in the purified polymer. Galacturonic and glucuronic acids, arabinose, xylose, mannose, and glucose are also present.     

Sloughing of cap cells and carbon exudation from maize seedling roots in compacted sand

Sloughing of root cap cells and exudation of mucilage plays an important role in the penetration of compacted soils by roots. For the first time we have quantified the rate of sloughing of root cap cells in an abrasive growth medium that was compacted to create mechanical impedance to root growth. The number of maize ( Zea mays ) root cap cells sloughed into sand increased as a result of compaction, from 1930 to 3220 d⁻¹ per primary root. This represented a 12-fold increase in the number of cells sloughed per mm root extension (from 60 to >700). We estimated that the whole of the cap surface area was covered with detached cells in compacted sand, compared with c . 7% of the surface area in loose sand. This lubricating layer of sloughed cells and mucilage probably decreases frictional resistance to soil penetration. The total carbon deposited by the root was estimated at c . 110 μg g⁻¹ sand d⁻¹. Sloughed cells accounted for <10% of the total carbon, the vast majority of carbon being contained in mucilage exudates.     

Contribution of root cap mucilage and presence of an intact root cap in maize (Zea mays) to the reduction of soil mechanical impedance

BACKGROUND AND AIMS: The impedance to root growth imposed by soil can be decreased by both mucilage secretion and the sloughing of border cells from the root cap. The aim of this study is to quantify the contribution of these two factors for maize root growth in compact soil. METHODS: These effects were evaluated by assessing growth after removing both mucilage (treatment I -- intact) and the root cap (treatment D -- decapped) from the root tip, and then by adding back 2 micro L of mucilage to both intact (treatment IM -- intact plus mucilage) and decapped (treatment DM -- decapped plus mucilage) roots. Roots were grown in either loose (0.9 Mg m(-3)) or compact (1.5 Mg m(-3)) loamy sand soils. Also examined were the effects of decapping on root penetration resistance at three soil bulk densities (1.3, 1.4 and 1.5 Mg m(-3)). KEY RESULTS: In treatment I, mucilage was visible 12 h after transplanting to the compact soil. The decapping and mucilage treatments affected neither the root elongation nor the root widening rates when the plants were grown in loose soil for 12 h. Root growth pressures of seminal axes in D, DM, I and IM treatments were 0.328, 0.288, 0.272 and 0.222 MPa, respectively, when the roots were grown in compact soil (1.5 Mg m(-3) density; 1.59 MPa penetrometer resistance). CONCLUSIONS: The contributions of mucilage and presence of the intact root cap without mucilage to the lubricating effect of root cap (percentage decrease in root penetration resistance caused by decapping) were 43 % and 58 %, respectively. The lubricating effect of the root cap was about 30 % and unaffected by the degree of soil compaction (for penetrometer resistances of 0.52, 1.20 and 1.59 MPa).     

Cell wall formation in zoospores of Allomyces arbuscula

Carbohydrate composition was determined in isolated cell walls of meiospores of Allomyces arbuscula after incubation for 15 min (encysted meiospores: cysts), 150 min (germlings: cysts + rhizoids) and 24 h (cysts + rhizoids + hyphae). The principal constituent in all cell wall samples is chitin, accounting for about 75% of the recovered carbohydrates. In addition, cell walls of all stages examined contain polysaccharides which release galactose, glucose, mannose, arabinose, xylose, fucose, and rhamnose on acid hydrolysis. While different developmental stages show minor quantitative changes in chitin, the ratio of galactose to glucose decreases sharply during differentiation of ungerminated cysts into germlings with rhizoids and hyphae. The increase in glucose is accompanied by a decrease in the amount of xylose and/or fucose and of galactose.List of Abbreviation TFA trifluoroacetic acid     

The impact of supplemental carbon sources on <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> growth,chlorophyll content and anthocyanin accumulation

This research explores the impacts of a broad range of supplemental carbon sources on growth and development of Arabidopsis thaliana. Parameters measured include dark-germinated hypocotyl length, light-germinated root growth, rosette growth, chlorophyll concentration and anthocyanin content. Treatment sugars include sucrose, maltose, d-glucose, d-fructose, l-arabinose, l-fucose, d-galactose, d-mannose, l-rhamnose and d-xylose each supplied at 4, 20 or 100 mM. This comparison of the effect of different carbon sources on multiple parameters and under identical conditions showed that every carbon source had unique qualitative and quantitative effects on Arabidopsis growth and development. Root growth was particularly sensitive to supplemental carbon source. Growth on 100 mM sucrose, maltose, glucose or xylose stimulated root growth by ~100%. Growth on arabinose, fucose, galactose, mannose or rhamnose inhibited root growth by 50% or more. Several sugars that strongly inhibited root growth had either no effect (galactose and fucose) or a positive effect (arabinose) on hypocotyl elongation and rosette growth. Rhamnose was the only carbon source that inhibited hypocotyl elongation across all concentrations. Sucrose, maltose, glucose, fructose, arabinose or xylose stimulated rosette growth by ~50%. Chlorophyll content was strongly reduced by mannose while sucrose, glucose, galactose and rhamnose caused smaller reductions. Anthocyanin accumulation was strongly induced by both galactose and mannose. Only mannose impacted all parameters across all concentrations. Based on these data it can be concluded that the effect of each carbon source on Arabidopsis growth and development is specific in terms of both magnitude and the parameters impacted.     

An Examination of Centrifugation as a Method of Extracting an Extracellular Solution from Peas, and Its Use for the Study of Indoleacetic Acid-induced Growth

A technique of centrifuging pea epicotyl sections which extracts water-soluble cell wall polysaccharides with less than 1.5% cytoplasmic contamination as revealed by malate dehydrogenase activity determinations was developed. Tests for protein, hexose, pentose, and malate dehydrogenase indicate that significant damage to the cells occurs above 3,000g. Below this force, there is little damage, as evidenced by the similar growth rates of centrifuged and noncentrifuged sections. Centrifugation at 1,000g extracts polysaccharides containing rhamnose, fucose, arabinose, xylose, mannose, galactose, and glucose. An increase in xylose and glucose, presumably xyloglucan, is induced by treating sections with indoleacetic acid. Much of the alcohol-insoluble, water-soluble polysaccharide within the wall is extractable by centrifugation, since nearly as much arabinose and xylose are extractable by centrifugation as by homogenization. The utility of this method for the study of cell wall metabolism is discussed.     

Synthesis and application of Fmoc-hydrazine for the quantitative determination of saccharides by reversed-phase high-performance liquid chromatography in the low and subpicomole range

A new derivatization reagent, Fmoc-hydrazine, has been synthesized from the reaction of Fmoc-chloroformate with hydrazine as a precolumn fluorometric labeling reagent for reducing sugars such as glucose, galactose, mannose, fructose, fucose, ribose, xylose, arabinose, lactose, and maltose. The optimization of derivatization conditions was examined in detail. Using a reversed-phase high-performance C-8 column and a mobile phase consisting of acetonitrile-aqueous acetic acid, seven sugar derivatives were separated under either isocratic or gradient conditions within 20 min. The Fmoc-hydrazine and sugar Fmoc-hydrazone derivatives exhibit excellent stability. The extent of the hydrazone formation was 77 and 82% for mannose and fucose as assessed by Dionex high-performance anion-exchange chromatography with pulsed amperometric detection. Linear calibration graphs were established in the range from 0.5 to 2 pmol and 12 to 110 pmol for individual sugar derivatives. The determination limits were 0.05-0.09 pmol for mannose, galactose, and ribose; 0.1 pmol for maltose, xylose, and glucose; 0.2 pmol for fucose and lactose; 0.3 pmol for arabinose; and 0.4 pmol for fructose. The component monosaccharides of ultramicroquantities of two glycoproteins (e.g., from 7 ng fetuin and ovalbumin) were determined in the subpicomole range.     

Physical properties of axenic maize root mucilage

Root mucilage was collected from 3–4 day-old axenically-grown maize seedlings (Zea mays L. cv. Freya). The water potential of the hydrated mucilage was measured by thermocouple psychrometry and the rheology at low deformation rates was studied using an oscillating cone and plate rheometer which provides information on both the elastic and viscous components of its behaviour. Water potential decreased as mucilage solute concentration increased, reaching a value of −60kPa at 1.2 mg mL⁻¹. At the lowest oscillation rate, the mucilage had a dynamic viscosity of 145 mPa s and behaved as a weak viscoelastic gel. After filtration to remove suspended root cap cells and other solid plant material, mucilage viscosity was reduced to 5–10 mPa s at low oscillation rates and the behaviour was that of a viscous liquid. The decrease in viscosity which occurs on filtration indicates that the root cap cells form an integral part of the gel system, either by interacting directly with each other or via the polysaccharide. Our observations provide further support for the idea that mucilage plays a major role in maintaining root-soil contact in the rhizosphere. This revised version was published online in June 2006 with corrections to the Cover Date.     

中性糖在土壤中的来源与分布特征

糖类(即碳水化合物)是土壤有机质的重要组成部分, 经生物化学降解形成不同结构的单糖。土壤中的中性单糖也叫中性糖, 主要包括木糖、核糖、阿拉伯糖、葡萄糖、半乳糖、甘露糖、岩藻糖和鼠李糖。其中, 植物来源的糖主要为五碳糖, 如木糖和阿拉伯糖; 微生物来源的糖主要包括半乳糖、甘露糖、岩藻糖、鼠李糖等六碳糖。研究中常利用六碳糖和五碳糖的比例指示微生物和植物对土壤有机碳的相对贡献。中性糖是微生物重要的碳源和能量来源, 在团聚体的形成过程中扮演着重要角色。该文整合了近30年土壤中性糖的研究进展, 对比了提取中性糖的常用方法, 分析了不同土地利用类型和不同土壤组分中中性糖的含量、来源和周转特征, 综述了影响中性糖含量和分布的主要环境因素。结果表明, 中性糖在耕地土壤中的绝对含量和相对含量均显著低于针叶林、阔叶林、草地和灌丛4种土地利用类型。(半乳糖+甘露糖)/(阿拉伯糖+木糖)(GM/AX)在不同土地利用间差异不显著, 而(鼠李糖+岩藻糖)/(阿拉伯糖+木糖)(RF/AX)则表明草地土壤中的微生物来源的中性糖含量高于针叶林和耕地。不同密度的土壤组分中, 轻质组分中中性糖的含量比重质组分高, 重质组分中微生物来源的中性糖较多; 就不同粒径(或团聚体)而言, 黏粒(或微团聚体)中微生物来源的中性糖含量更丰富。有关影响土壤中性糖含量和分布的因素的研究, 目前主要集中在人为活动(如耕种和放牧等), 而有关温度、降水等自然环境因素影响的研究较少。     

Aquilinan,an acidic polysaccharide from Pteridium aquilinum

A water-soluble, acidic polysaccharide, aquilinan, was isolated from storage rhizomes and young fronds of bracken and shown to be homogeneous. The polysaccharide contained galactose, xylose, fucose and arabinose residues together with chains of repeating α(1-2′)glucuronosylmannose units. A method of correcting the mannose content for degradation during the hydrolysis of the aldobiuronic acid is described.     

Fucose in the surface deposits of axenic and field grown roots ofZea mays L.

Summary The location of materials containing terminal fucose residues on the surface of axenic and field grown roots of corn has been determined.Binding patterns of FITC-labelled,Lotus purpureus Moench lectin indicate the presence of the fucose residues in the cell walls and mucilage of the peripheral region of the root cap. During development, fucose residues also appear in the outer periclinal walls and overlying mucilage of columnar epidermal cells. Surface material rich in these residues persists between the mature root hairs but is not found on their surface. Fucose-rich mucilage is present on the exposed surface of aerial roots and at the point where they enter the soil. No lectin binding residues are indicated elsewhere in the roots.