不同硼效率甘蓝型油菜品种细胞壁中硼的分配

应用不同硼效率甘蓝型油菜品种 ,研究硼在细胞壁中的分配。硼主要结合在细胞壁中 ,缺硼显著提高硼在细胞壁中的分配比例。根系细胞壁硼含量显著低于叶片 ,但根系细胞壁硼占根系总硼量之比例显著高于叶片。同一品种根系及其细胞壁、老叶细胞壁硼含量受生育期影响较小 ,新叶及其细胞壁、老叶硼含量受生育期影响较大。在正常供硼条件下 ,硼高效品种根系细胞壁和叶片细胞壁硼含量均低于低效品种 ;正常和缺硼条件下 ,硼高效品种细胞壁硼占器官总硼量之比例均低于低效品种。说明硼低效品种需较多的硼构建细胞壁。     

Localization of Boron in Cell Walls of Squash and Tobacco and Its Association with Pectin (Evidence for a Structural Role of Boron in the Cell Wall)

B deficiency results in a rapid inhibition of plant growth, and yet the form and function of B in plants remains unclear. In this paper we provide evidence that B is chemically localized and structurally important in the cell wall of plants. The localization and chemical fractionation of B was followed in squash plants (Curcurbita pepo L.) and cultured tobacco cells (Nicotiana tabacum) grown in B-replete or B-deficient medium. As squash plants and cultured tobacco cells became deficient, an increasingly large proportion of cellular B was found to be localized in the cell wall. Cytoplasmic B concentrations were reduced to essentially zero as plants became deficient, whereas cell wall B concentration remained at or above 10 [mu]g B/g cell wall dry weight in all experiments. Chemical and enzymic fractionation studies suggest that the majority of cell B is associated with pectins within the cell wall. Physical analysis of B-deficient tissue indicates that cell wall plastic extensibility is greatly reduced under B deficiency, and anatomical observations indicate that B deficiency impairs normal cell elongation in growing plant tissue. In plants in which B deficiency had inhibited all plant growth, tissues remained green and did not show any additional visible symptoms for at least 1 week with no additional B. This occurred even though cytoplasmic B had been reduced to extremely low levels (<0.2 [mu]g/g). This suggests that B in these species is largely associated with the cell wall and that any cytoplasmic role for B is satisfied by very low concentrations of B. The localization of B in the cell wall, its association with cell wall pectins, and the contingent effects of B on cell wall extensibility suggest that B plays a critical, although poorly defined, role in the cell wall structure of higher plants.     

Inhibitory Effects of a Pectin-Enriched Tomato Cell Wall Fraction on Agrobacterium tumefaciens Binding and Tumor Formation

A pectin-enriched soluble cell wall fraction (CWF) prepared from suspension cultured tomato cells inhibits binding of Agrobacterium tumefaciens to these cells. It was hypothesized that the CWF contains the plant surface binding site for A. tumefaciens (NT Neff, AN Binns 1985 Plant Physiol 77: 35-42). Experiments described here demonstrate that tomato CWF inhibited tumor formation on potato slices and Agrobacterium binding to intact tomato cells in a dose-dependent fashion. Boiling the fraction reduced both its binding and tumor inhibitory activities. Tumor inhibitory activity was titrated out by increased concentrations of bacterial inocula with no inhibition apparent at 1 × 10⁸ bacteria per milliliter. These results indicate that a tomato CWF is enriched for a putative A. tumefaciens binding site which may also be involved in tumor formation in potato.     

Changes in the Distribution of Pectin Methylesterase across Leaf Abscission Zones of Phaseolus vulgaris

Determinations of pectin methylesterase in abscission zonesand surrounding parts of Phaseolus leaves have shown a relativelyhigh activity in young tissues, which decreases as the leafgrows older. The greatest fall occurs in the pulvinus. In non-senescent leaves, the enzyme activity is higher in pulvinithan elsewhere and there is a gradient of activity across theline of abscission from the pulvinus to the petiole or stem.During senescence this gradient falls, and is generally reversedat abscission. Data suggest that the advent of abscission maybe linked to the steepness of this gradient. If abscission of leaves or isolated abscission zones is acceleratedby appropriate treatments there is a more rapid decrease inpectin methylesterase activity than in the controls, and a morerapid fall in the gradient across the line of abscission. Ifabscission is retarded the enzyme activity is sustained andthe gradient is maintained or increased. The results are discussed in relation to the changes that mayoccur in the pectic constituents of cell walls during senescenceand abscission, and in relation to some of the known effectsof auxins on pectin metabolism.     

Cell Wall Metabolism in Ripening Fruit (VII. Biologically Active Pectin Oligomers in Ripening Tomato (Lycopersicon esculentum Mill.) Fruits)

A water-soluble, ethanol-insoluble extract of autolytically inactive tomato (Lycopersicon esculentum Mill.) pericarp tissue contains a series of galacturonic acid-containing (pectic) oligosaccharides that will elicit a transient increase in ethylene biosynthesis when applied to pericarp discs cut from mature green fruit. The concentration of these oligosaccharides in extracts (2.2 [mu]g/g fresh weight) is in excess of that required to promote ethylene synthesis. Oligomers in extracts of ripening fruits were partially purified by preparative high-performance liquid chromatography, and their compositions are described. Pectins were extracted from cell walls prepared from mature green fruit using chelator and Na2CO3 solutions. These pectins are not active in eliciting ethylene synthesis. However, treatment of the Na2CO3-soluble, but not the chelator-soluble, pectin with pure tomato polygalacturonase 1 generates oligomers that are similar to those extracted from ripening fruit (according to high-performance liquid chromatography analysis) and are active as elicitors. The possibility that pectin-derived oligomers are endogenous regulators of ripening is discussed.     

Ionic Diffusion Through the Nitella Cell Wall in the Presence of Calcium

Diffusion potentials (concentration and bi- or multi-ionic potentials)in KC1, NaCl, or LiCl solutions have been measured across anisolated cell wall of Nitella, with or without the same concentrationof CaCl² on either side of the cell wall. The absolute valueof the potentials decreases as the external Ca²⁺concentrationincreases and it may happen that an inversion of the sign ofthe concentration potentials results when the external Ca²⁺solution reaches 1 mM. Dosages of K⁺ and Ca²⁺ in the cell wallhave shown that Ca²⁺ easily displaces the monovalent ion fromthe exchange sites and tends to neutralize the cationic exchanger.However, in most cases, the measured potentials are still morenegative than the theoretical potentials which would be setup by a neutral-site membrane in the same conditions. Theseresults suggest that Ca²⁺ largely reduces the discriminationproperties of the cell wall between cations and anions.     

硼缺乏导致花粉管细胞壁多糖分布的改变

集应用免疫细胞化学及显微红外光谱分析技术,深入研究了硼元素对花粉管生长的调节作用。用识别甲酯化果胶的单克隆抗体JIM7和识别酸性果胶的JIM5对离体培养的百合（LiliumlongiflorumThunb．）及烟草（NicotianatabacumL.cv．“PetitHavana”）花粉管进行免疫荧光标记,发现无硼培养导致细胞壁果胶成分呈异常分布,酸性果胶在花粉管顶端大量富集;苯胺蓝诱导荧光法揭示,无硼培养引起胼胝质在顶端细胞壁积累。通过显微红外光谱（FTIR）分析,进一步验证了无硼培养导致细胞壁酸性果胶质含量的增加,并发现酚酯含量比正常情况减少,而游离酚类化合物含量明显增加。上述结果表明,硼可能作为一种相关因子影响关键酶活性,改变细胞壁多糖网状结构以至细胞壁的延展性,从而调节花粉管生长;酚类积累对质膜完整性的影响也会对调节花粉管生长有间接作用。     

An Antisense Pectin Methylesterase Gene Alters Pectin Chemistry and Soluble Solids in Tomato Fruit

Pectin methylesterase (PME, EC 3.1.11) demethoxylates pectins and is believed to be involved in degradation of pectic cell wall components by polygalacturonase in ripening tomato fruit. We have introduced antisense and sense chimeric PME genes into tomato to elucidate the role of PME in fruit development and ripening. Fruits from transgenic plants expressing high levels of antisense PME RNA showed <10% of wild-type PME enzyme activity and undetectable levels of PME protein and mRNA. Lower PME enzyme activity in fruits from transgenic plants was associated with an increased molecular weight and methylesterification of pectins and decreased levels of total and chelator soluble polyuronides in cell walls. The fruits of transgenic plants also contained higher levels of soluble solids than wild-type fruits. This trait was maintained in subsequent generations and segregated in normal Mendelian fashion with the antisense PME gene. These results indicate that reduction in PME enzyme activity in ripening tomato fruits had a marked influence on fruit pectin metabolism and increased the soluble solids content of fruits, but did not interfere with the ripening process.     

The Tomato Fruit Cell Wall : II. Polyuronide Metabolism in a Nonsoftening Tomato Mutant

A nonsoftening tomato (Lycopersicon esculentum L.) variety, dg, was examined to assess the physiological basis for its inability to soften during ripening. Total uronic acid levels, 18 milligrams uronic acid/100 milligrams wall, and the extent of pectin esterification, 60 mole%, remained constant throughout fruit development in this mutant. The proportion of uronic acid susceptible to polygalacturonase in vitro also remained constant. Pretreatment of heat-inactivated dg fruit cell walls with tomato pectinmethylesterase enhances polygalacturonase susceptibility at all ripening stages. Pectinesterase activity of cell wall protein extracts from red ripe dg fruit was half that in extracts from analogous tissue of VF145B. Polygalacturonase activities of cell wall extracts, however, were similar in both varieties. Diffusion of uronic acid from tissue discs of both varieties increased beginning at the turning stage to a maximum of 2.0 milligrams uronic acid released/gram fresh weight at the ripe stage. The increased quantity of hydrolytic products released during ripening suggests the presence of in situ polygalacturonase activity. Low speed centrifugation was employed to induce efflux of uronide components from the cell wall tree space. In normal fruit, at the turning stage, 2.1 micrograms uronic acid/gram fresh weight was present in the eluant after 1 hour, and this value increased to a maximum of 8.2 micrograms uronic acid/gram fresh weight at the red ripe stage. However, centrifuge-aided extraction of hydrolytic products failed to provide evidence for in situ polygalacturonase activity in dg fruit. We conclude that pectinesterase and polygalacturonase enzymes are not active in situ during the ripening of dg fruit. This could account for the maintenance of firmness in ripe fruit tissue.     

Posttranslational Elevation of Cell Wall Invertase Activity by Silencing Its Inhibitor in Tomato Delays Leaf Senescence and Increases Seed Weight and Fruit Hexose Level

Invertase plays multiple pivotal roles in plant development. Thus, its activity must be tightly regulated in vivo. Emerging evidence suggests that a group of small proteins that inhibit invertase activity in vitro appears to exist in a wide variety of plants. However, little is known regarding their roles in planta. Here, we examined the function of INVINH1, a putative invertase inhibitor, in tomato (Solanum lycopersicum). Expression of a INVINH1:green fluorescent protein fusion revealed its apoplasmic localization. Ectopic overexpression of INVINH1 in Arabidopsis thaliana specifically reduced cell wall invertase activity. By contrast, silencing its expression in tomato significantly increased the activity of cell wall invertase without altering activities of cytoplasmic and vacuolar invertases. Elevation of cell wall invertase activity in RNA interference transgenic tomato led to (1) a prolonged leaf life span involving in a blockage of abscisic acid–induced senescence and (2) an increase in seed weight and fruit hexose level, which is likely achieved through enhanced sucrose hydrolysis in the apoplasm of the fruit vasculature. This assertion is based on (1) coexpression of INVINH1 and a fruit-specific cell wall invertase Lin5 in phloem parenchyma cells of young fruit, including the placenta regions connecting developing seeds; (2) a physical interaction between INVINH1 and Lin5 in vivo; and (3) a symplasmic discontinuity at the interface between placenta and seeds. Together, the results demonstrate that INVINH1 encodes a protein that specifically inhibits the activity of cell wall invertase and regulates leaf senescence and seed and fruit development in tomato by limiting the invertase activity in planta.     

Ultrastructure of the Cell Wall of Vessel Contact Cells in the Xylem of Tomato Stems

Mature contact cells in the metaxylem of tomato stems are characterizedby a distinctive wall deposit occurring mainly over the areaof the pit membrane that separates the cell from an adjoiningvessel member but also extending as a layer on the secondarywall round the interior of the cell. Cytochemical tests showthe layer contains a low concentration of pectin, no lignin,and a high concentration of polysaccharides. It can be distinguishedfrom the primary and secondary walls by quantitative differencesin the cytochemical staining reactions and by its loose-texturedappearance under the electron microscope. The layer is similarin composition and structure to the ‘protective layer’described in woody plants. The layer has also been found inpea and cotton and may be a characteristic feature of the contactcells in herbaceous plants. Cell wall ultrastructure, contact cells, Lycopersicon esculentum, Mill., protective layer, tomato, xylem     

Calcium Antagonist TMB-8 Inhibits Cell Wall Formation and Growth in Pea

The effects on auxin-stimulated growth and cell wall formationof 8-(N, N-diethylamino)-octyl-3, 4, 5-trimethoxybenzoate.HCI(TMB-8), an intracellular Ca²⁺ antagonist, were investigatedin abraded stem segments from aetiolated seedlings of Pisumsativum L. cv. Alaska. Incubation of segments at pH 6.0 with200 mmol m^–3 TMB-8 resulted in a 50% inhibition of auxin-stimulatedgrowth. Added Ca²⁺ did not restore normal auxin-stimulated growth,presumably because of its well-known stiffening effect on thecell wall. In segments incubated at a pH (7–2) which preventedelongation, auxin promoted the incorporation of [³H]glucoseinto the cell wall relative to total uptake of label. TMB-8abolished about 60% of the total incorporation of label intocell walls in the presence of auxin, but was not effective inthe absence of auxin. Exogenous CaCl₂ reversed the inhibitoryeffect of TMB-8 on relative cell wall incorporation in a parabolicmanner, with a 50% reversal at about 100 mmol m^–3 andcomplete reversal at 1.0 mol m^–3 Ca²⁺. Other ions tested(Mg²⁺, Mn²⁺, Cu²⁺, Zn²⁺) were without substantial effect atconcentrations of 0.5 mol m^–3. Both apparent uptake ofCa²⁺ and consequent reversal of TMB-8 inhibition of cell wallincorporation were blocked by the Ca²⁺ channel blockers verapamiland La³⁺. The data provide further evidence that auxin-stimulatedgrowth is dependent upon continued cell wall incorporation,and suggest that a Ca²⁺ messenger system may be involved inthe promotory actions of auxin on cell wall synthesis and long-termgrowth. Key words: Auxin, calcium, cell wall synthesis     

Pectin Demethylesterification Generates Platforms that Anchor Peroxidases to Remodel Plant Cell Wall Domains

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Changes in Tomato Leaves Induced by NaCl Stress: Leaf Organization and Cell Ultrastructure

The alterations of organization of leaf tissues and cell ultrastructure as a consequence of salt stress (75 and 150 mM NaCl) were studied in two tomato (Lycopersicum esculentum Mill.) cultivars showing different salinity tolerance. The salinity brought changes in cell shape, volume of intercellular spaces and chloroplast number, shape and size. These characteristics were specific in each cultivar. The ultrastructural changes were also different in the two tomato cultivars studied and the most important ones were in the number and size of starch granules in chloroplasts, the number of electron-dense corpuscules in the cytoplasm, the structure of mitochondria, and number of plastoglobuli. This revised version was published online in July 2006 with corrections to the Cover Date.     

Calcium in the Root-hair Wall

The walls of normal root hairs of white mustard, corn and tomatoseedlings developed in moist air following seed treatment with⁴⁵CaCl₂, were tested by radioautography for incorporated calcium.Radioactivity was absent or faint over the growing tips butbecame concentrated immediately behind this point. It extendedback along the walls of the hairs, increasing in concentrationtowards the base. Since the hairs were subjected to plasmolysisand prolonged extraction with hot water prior to making theradioautographs, the distribution of radioactivity indicatedthe presence of bound ⁴⁵Ca. These findings are compatible withthe view that gradual calcification is essential for the normalgrowth and form of the hair. Radioautographs of mustard root hairs produced in an aqueoussolution of ⁴⁵CaCl₂ showed conspicuous radioactivity throughoutthe whole length of the hairs but particularly over the tips.Excessive wall calcification at the tips of these hairs couldbe the cause of their stunted growth.     

Rapid Effects of Boron Deficiency on Cell Wall Elasticity Modulus in Cucurbita pepo Roots

Rinsing roots of Cucurbita pepo with an essentially boron-free nutrient solution results in a significant reduction of the cell wall elasticity modulus as determined by the cell presure probe (measuring always in the same cells). This points to a role of boron in crosslinking cell wall macromolecules. Longer exposure to boron deficient conditions, however, increased again and above the initial values. This trend became visible between 20 and 30 minutes after starting the deficiency treatment. There is no indication for a loss of turgor during the time course of the experiments, suggesting that boron primarily affects cell wall physical properties and that membrane leakiness, as observed by other groups, is a secondary event. Several testable hypotheses for the action of boron are presented.     

Changes in Pectin Structure during Epidermal Cell Elongation in Pea (Pisum sativum) and Its Implications for Cell Wall Architecture

Changes in cell wall architecture during elongation of epidermalcells in pea epicotyls were visualized by rapid-freezing anddeep-etching (RFDE) techniques. The abundant network structurecomposed of the association of granular substances disappearedfrom the cell wall during elongation. The granular substanceswere demonstrated to be pectic polysaccharides by their disappearanceupon EDTA treatment and by chemical analysis of the EDTA-ex-tractablesubstances. Labeling with the monoclonal antibody JIM5, whichrecognizes unesterified pectins, was much more extensive inthe cell walls of the non-elongating region than in those ofthe elongating region. The pore size of the cell wall was largerin the non-elongating region than in the elongating region.These observations suggest that the formation of the pecticgel itself is not involved in the control of the wall porosity.We proposed that the association of the granular substancesis involved in the swelling of the cell walls in the elongatingregion. (Received May 18, 1998; Accepted September 25, 1998)     

Water Stress and Foliar Boron Application Altered Cell Wall Boron and Seed Nutrition in Near-Isogenic Cotton Lines Expressing Fuzzy and Fuzzless Seed Phenotypes

Our previous research, conducted under well-watered conditions without fertilizer application, showed that fuzziness cottonseed trait resulted in cottonseed nutrition differences between fuzzy (F) and fuzzless (N) cottonseed. Under water stress conditions, B mobility is further limited, inhibiting B movement within the plant, affecting seed nutrition (quality). Therefore, we hypothesized that both foliar B and water stress can affect B mobility, altering cottonseed protein, oil, and mineral nutrition. The objective of the current research was to evaluate the effects of the fuzziness seed trait on boron (B) and seed nutrition under water stress and foliar B application using near-isogenic cotton lines (NILs) grown in a repeated greenhouse experiment. Plants were grown under-well watered conditions (The soil water potential was kept between -15 to -20 kPa, considered field capacity) and water stress conditions (soil water potential between -100 and -150 kPa, stressed conditions). Foliar B was applied at a rate of 1.8 kg B ha^-1 as H₃BO₃. Under well-watered conditions without B the concentrations of seed oil in N lines were higher than in F lines, and seed K and N levels were lower in N lines than in F lines. Concentrations of K, N, and B in leaves were higher in N lines than in F lines, opposing the trend in seeds. Water-stress resulted in higher seed protein concentrations, and the contribution of cell wall (structural) B to the total B exceeded 90%, supporting the structural role of B in plants. Foliar B application under well-watered conditions resulted in higher seed protein, oil, C, N, and B in only some lines. This research showed that cottonseed nutrition differences can occur due to seed fuzziness trait, and water stress and foliar B application can alter cottonseed nutrition.     

Distribution of Yeast Cell Wall Lytic Activity among Microorganisms

An α-glucosidase has been isolated from the mycelia of Penicillium purpurogenum in electrophoretically homogeneous form, and its properties have been investigated. The enzyme had a molecular weight of 120,000 and an isoelectric point of pH 3.2. The enzyme had a pH optimum at 3.0 to 5.0 with maltose as substrate. The enzyme hydrolyzed not only maltose but also amylose, amylopectin, glycogen, and soluble starch, and glucose was the sole product from these substrates. The Km value for maltose was 6.94×10^?4 m. The enzyme hydrolyzed phenyl α-maltoside to glucose and phenyl α-glucoside. The enzyme had α-glucosyltransferase activity, the main transfer product from maltose being maltotriose. The enzyme also catalyzed the transfer of α-glucosyl residue from maltose to riboflavin.