Tunicamycin Prevents Cellulose Microfibril Formation in Oocystis solitaria

The effect of tunicamycin (TM) on the development of the cell wall in Oocystis solitaria has been investigated. It was found that 10 micromolar TM completely stops the assembly of new microfibrils as observed at the ultrastructural level. During cell wall formation, freeze fracture replicas of the E-face of the plasma membrane reveal two major substructures: the terminal complexes (TC), paired and unpaired, and the microfibril imprints extending from unpaired TCs. In cells treated for 3 hours or longer with TM, the TCs are no longer visible, whereas microfibril imprints are still present. Because of the reported highly selective mode of action of TM, our results implicate a role for lipid-intermediates in cellulose synthesis in O. solitaria. It is assumed that TM prevents the formation of a glycoprotein which probably is a fundamental part of the TCs and may act as a primer for the assembly of the microfibrils.     

Evidence for the Regulation of the Shift in Cellulose Microfibril Orientation in Freeze-Fractured Plasma Membrane of Boergesenia forbesii

Morphological evidence regarding the regulation of the directionof movement of the cellulose-synthesizing complexes in Boergeseniawas obtained by observing the distribution of intramembranousparticles (IMPS) around the complexes. In complexes with straightconfigurations, the distribution of IMPS on both sides of thecomplexes was almost equal. In complexes characterized by acurved configuration, however, the IMPS density was greaterin the outer half-circle of the curve. Some particles on thisside were attached to or intruded into the complex, and theirsizes were no different from those of the IMPS on the fracturedplane. These results indicate that the distribution and orientationof the particles in relation to the complex have been affectedby membrane fluidity and, that membrane fluidity might playa key role in determining the regulation of the direction ofmovement of the complex. (Received July 16, 1984; Accepted October 12, 1984)     

Diversity of Cellulose Microfibril Arrangement in the Cell Walls of Lygeum spartum Leaves

Extraction of matrix material, cytochemical methods and stereoanalysiswere used to study cell-wall architecture in leaves of Lygeumspartum, a grass species growing in sub-and regions of the Mediterraneanzone Most tissues were lignified with the exception of mesophyllcells Better results were obtained with NaOH extraction thanwith methylamine extraction The different types of fibres (sub-epidermal,axial and bundle sheath fibres), parenchyma and epidermal cellswere characterized by specific cell-wall organization The differentwall textures were variations of the basic helicoidal model Lygeum spartum L, lignified leaf fibres, cell wall, helicoidal texture, polysaccharide cytochemistry     

Orientation of Wall Microfibril Deposition in Root Cells of Pisum sativum L. var Alaska

The arrangement of wall microfibrils on the inner surface ofcortical cells in pea roots was observed by a replica method.In the elongating region, microfibrils were deposited transverselyto the root axis. After cell elongation stopped, the orientationof microfibril deposition changed discontinuously from a transverseto an oblique one. The change occurred at 6–7 mm fromthe root tip. The oblique orientation seemed to change discontinuouslyto another oblique one as time passed. (Received January 24, 1986; Accepted May 15, 1986)     

Loosening Xyloglucan Accelerates the Enzymatic Degradation of Cellulose in Wood

In order to create trees in which cellulose, the most abundant component in biomass, can be enzymatically hydrolyzed highly for the production of bioethanol, we examined the saccharification of xylem from several transgenic poplars, each overexpressing either xyloglucanase, cellulase, xylanase, or galactanase. The level of cellulose degradation achieved by a cellulase preparation was markedly greater in the xylem overexpressing xyloglucanase and much greater in the xylems overexpressing xylanase and cellulase than in the xylem of the wild-type plant. Although a high degree of degradation occurred in all xylems at all loci, the crystalline region of the cellulose microfibrUs was highly degraded in the xylem overexpressing xyloglucanase. Since the complex between microfibrils and xyloglucans could be one region that is particularly resistant to cellulose degradation, loosening xyloglucan could facilitate the enzymatic hydrolysis of cellulose in wood.     

Effects of Light and Inhibitors on Polylamellation and Shift of Microfibril Orientation in Boergesenia Cell wall

New round cells of Boergesenia forbesii, which develop fromprotoplasts, form crossed polylamellate cell walls. The frequencyof the shift in microfibril orientation in generating wall layerswas affected not by the photoperiod, but by the total lengthof the illumination period. The dependency of the shift uponlight intensity and quality, and its inhibition by 3-(3,4-dichlorophenyl)-l,l-dimethylureaindicated that the shift in fibril orientation was primarilydependent upon photosynthesis. KCN and NaN₃ strongly inhibitedwall thickening but the frequency of the shift only a little.Therefore, the system controlling microfibril orientation functionsindependently of wall synthesis. Both colchicine and cytochalasinB did not affect microfibril orientation as well as wall synthesisin Boergesenia cells. (Received August 1, 1981; Accepted December 14, 1981)     

Microfibril Orientation Dominates the Microelastic Properties of Human Bone Tissue at the Lamellar Length Scale

The elastic properties of bone tissue determine the biomechanical behavior of bone at the organ level. It is now widely accepted that the nanoscale structure of bone plays an important role to determine the elastic properties at the tissue level. Hence, in addition to the mineral density, the structure and organization of the mineral nanoparticles and of the collagen microfibrils appear as potential key factors governing the elasticity. Many studies exist on the role of the organization of collagen microfibril and mineral nanocrystals in strongly remodeled bone. However, there is no direct experimental proof to support the theoretical calculations. Here, we provide such evidence through a novel approach combining several high resolution imaging techniques: scanning acoustic microscopy, quantitative scanning small-Angle X-ray scattering imaging and synchrotron radiation computed microtomography. We find that the periodic modulations of elasticity across osteonal bone are essentially determined by the orientation of the mineral nanoparticles and to a lesser extent only by the particle size and density. Based on the strong correlation between the orientation of the mineral nanoparticles and the collagen molecules, we conclude that the microfibril orientation is the main determinant of the observed undulations of microelastic properties in regions of constant mineralization in osteonal lamellar bone. This multimodal approach could be applied to a much broader range of fibrous biological materials for the purpose of biomimetic technologies.     

Microfibril formation in chick notochordal cells

The central parts of the chick notochord at Hamburger and Hamilton's stages 20–22 were investigated by electron microscopy. Electron-dense bodies of various sizes and shapes and bounded by a limiting membrane were found in the central cells of the notochord. These dense bodies contained fibrous material or microfibrils which ranged from 120 to 600 Å in diameter. The large microfibrils often exhibited a typical repeating period with an interval of about 320 Å. These dense bodies were always located near the cell membrane, which is rough or irregular in the central parts of the notochord at these stages. The fibrous core material of the dense body frequently shows striking similarities to amorphous fibrous material in the intercellular space of the central parts of the notochord, where they are situated at a considerable distance from the perinotochordal sheath space. From these results, it seems reasonable to suggest that the central cells as well as the peripheral cells of the notochord are capable of forming microfibrils similar to those observed in the perinotochordal sheath space.Moreover, they may play an important role in the total fibrillogenesis of the notochord.     

Microfibril orientation in plant cell walls

Summary The distribution of particles on the surface of the plasmalemma in the collenchyma of Apium graveolens was studied by the freeze-etching technique. The aim was to determine whether the distribution of particles was related to the known longitudinal or transverse orientation of cellulose microfibrils in different layers of the walls of these cells. Preliminary statistical studies have shown no obvious correlation between particle distribution and microfibril orientation although the distribution appeared uniform rather than random. Qualitatively, the particle distribution on the plasmalemma of differentiating xylem fibres of Eucalyptus maculata and of the cortical parenchyma of Avena sativa coleoptiles appeared to be similar to that observed on the plasmalemma of Apium. No correlation between the particle distribution and the microfibril orientation known to exist in the walls of these cells could be discerned.The orientation of microtubules in the cytoplasm of collenchyma cells of Apium graveolens was parallel to the microfibril orientation in many instances, but exceptions were noted. A possible interpretation for this variation is discussed. It is concluded that the microtubules are the structures which are most likely to be involved in determining microfibril orientation in the cell wall.     

Synthesis and Study of Copper-Containing Polymers of Microcrystalline Cellulose Sulfates from Larch Wood

For the first time, the synthesis of water-soluble copper-containing microcrystalline cellulose sulfates (Сu-MCS) has been performed by the ion exchange method. The composition of the products has been studied by chemical methods and X-ray spectral microanalysis. The copper content in the Сu-MCS samples was 12.6–14.1%. The absence of sodium in the resulting polymer indicates the complete substitution of the sodium cations by the copper cations in the sodium salt of MCC sulfate. The structure of the copper-containing sulfates of microcrystalline cellulose has been confirmed by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and electron paramagnetic resonance (EPR). According to the XRD method, Сu-MCS and Na-MCS have an amorphous structure in contrast to the original MCC samples, which have a high degree of crystallinity. The EPR data have demonstrated the formation of a pseudocrystalline structure of the copper-containing salt system in the Сu-MCS samples. As shown by atomic-force microscopy, the surface of the Сu-MCS films consists of homogeneous crystallites, which have a spherical or slightly extended form with the size of about 70 nm. The film surface is quite homogeneous in its phase composition and contains no impurities.

     

Xyloglucan Endotransglycosylase Activity, Microfibril Orientation and the Profiles of Cell Wall Properties Along Growing Regions of Maize Roots

A series of physical and chemical analyses were made on theexpanding zone of maize seedling roots grown in hydroponics.Comparison of longitudinal profiles of local relative elementalgrowth rate and turgor pressure indicated that cell walls becomelooser in the apical 5 mm and then tighten 5–10 mm fromthe root tip. Immersion of roots in 200 mol m^–3 mannitol(an osmotic stress of 0·48 MPa) rapidly and evenly reducedturgor pressure along the whole growing region. Growth was reducedto a greater extent in the region 5–10 mm from the roottip than in the apical region. This indicated rapid wall-looseningin the root tip, but not in the more basal regions. Following 24 h immersion in 400 mol m^–3 mannitol (an osmoticstress of 0·96 MPa) turgor had recovered to pre-stressedvalues. Under this stress treatment, growth was reduced in theregion 4–10 mm from the root tip, despite the recoveryof turgor, indicating a tightening of the wall. In the rootapex, local relative elemental growth rate was unchanged incomparison to control tissue, showing that wall properties herewere similar to the control values. Cellulose microfibrils on the inner face of cortical cell wallsbecame increasingly more parallel to the root axis along thegrowth profile of both unstressed and stressed roots. Orientationdid not correlate with the wall loosening in the apical regionof unstressed roots, or with the tightening in the region 5–10mm from the root tip following 24 h of osmotic stress. Longitudinal profiles of the possible wall-loosening enzymexyloglucan endotransglycosylase (XET) had good correspondencewith an increase in wall loosening during development. In thezone of wall tightening following osmotic stress, XET activitywas decreased per unit dry weight (compared with the unstressedcontrol), but not per unit fresh weight. Key words: Osmotic stress, turgor, growth, cell wall properties, microfibrils, XET     

Microfibril Structure Masks Fibrillin-2 in Postnatal Tissues

Fibrillin microfibrils are polymeric structures present in connective tissues. The importance of fibrillin microfibrils to connective tissue function has been demonstrated by the multiple genetic disorders caused by mutations in fibrillins and in microfibril-associated molecules. However, knowledge of microfibril structure is limited, largely due to their insolubility. Most previous studies have focused on how fibrillin-1 is organized within microfibril polymers. In this study, an immunochemical approach was used to circumvent the insolubility of microfibrils to determine the role of fibrillin-2 in postnatal microfibril structure. Results obtained from studies of wild type and fibrillin-1 null tissues, using monoclonal and polyclonal antibodies with defined epitopes, demonstrated that N-terminal fibrillin-2 epitopes are masked in postnatal microfibrils and can be revealed by enzymatic digestion or by genetic ablation of Fbn1. From these studies, we conclude that fetal fibrillin polymers form an inner core within postnatal microfibrils and that microfibril structure evolves as growth and development proceed into the postnatal period. Furthermore, documentation of a novel cryptic site present in EGF4 in fibrillin-1 underscores the molecular complexity and tissue-specific differences in microfibril structure.     

Microfibril deposition on cultured protoplasts ofVicia hajastana

Summary Cell wall regeneration by protoplasts fromVicia hajastana suspension cultures was investigated with Calcofluor White ST staining and platinum-palladium surface replicas. Microfibril deposition was initiated after 10–20 minutes of culture and within 20 hours protoplasts were covered with a heavy mat of microfibrils. The early stages of microfibril formation could not be detected with Calcofluor staining.Supported by the National Research Council of Canada, Grant A6304.Supported by Deutsche Forschungsgemeinschaft.     

A Microfibril Formation from Depolymerized Chitosan by N-Acetylation

The geometrical isomers of O-4-tert-butylcyclohexyl methylthiocarbamate were prepared by way of the corresponding dithiocarbonates. The O-trans-isomer was obtained from a mixture of cis- and trans-4-tert-butylcyclohexanols via an alkoxide formation involving biassed equilibration, whereas the O-cis isomer was from the cis-cyclohexanol via an alkoxide formation free from equilibration.

The acid-catalyzed rearrangement of the O-cis thiocarbamate gave a 1:3 mixture of S-cis- and S-trans-4-tert-butylcyclohexyl methylthiocarbamates, whereas that of the O-trans thiocarbamate afforded a 9:1 mixture of S-cis and S-trans products.

These results, together with the data from a study on the reactions of O-cis-3,3,5-tri-methylcyclohexyl methylthiocarbamate and the O-neopentyl analog, indicate that the acid-catalyzed rearrangement proceeded mostly through an S_N2 type of transalkylating mechanism to give the S-isomer in the case where the approach of nucleophiles was not sterically blocked, otherwise the reaction gave S_N1-type products.     

Extending the Microtubule/Microfibril Paradigm : Cellulose Synthesis Is Required for Normal Cortical Microtubule Alignment in Elongating Cells

The cortical microtubule array provides spatial information to the cellulose-synthesizing machinery within the plasma membrane of elongating cells. Until now data indicated that information is transferred from organized cortical microtubules to the cellulose-synthesizing complex, which results in the deposition of ordered cellulosic walls. How cortical microtubules become aligned is unclear. The literature indicates that biophysical forces, transmitted by the organized cellulose component of the cell wall, provide a spatial cue to orient cortical microtubules. This hypothesis was tested on tobacco (Nicotiana tabacum L.) protoplasts and suspension-cultured cells treated with the cellulose synthesis inhibitor isoxaben. Isoxaben (0.25–2.5 μm) inhibited the synthesis of cellulose microfibrils (detected by staining with 1 μg mL⁻¹ fluorescent dye and polarized birefringence), the cells failed to elongate, and the cortical microtubules failed to become organized. The affects of isoxaben were reversible, and after its removal microtubules reorganized and cells elongated. Isoxaben did not depolymerize microtubules in vivo or inhibit the polymerization of tubulin in vitro. These data are consistent with the hypothesis that cellulose microfibrils, and hence cell elongation, are involved in providing spatial cues for cortical microtubule organization. These results compel us to extend the microtubule/microfibril paradigm to include the bidirectional flow of information.     

植物纤维素合成酶基因和纤维素的生物合成

纤维素地球上最丰富的生物大分子和最重要的可再生资源,1996年克隆了第一个植物纤维素合成酶基因,植物纤维素的生物合成需要多个纤维素合成酶与其他相关酶如Korrigan纤维素酶,蔗糖合成酶等来共同完成。本文介绍了植物纤维素合成酶基因和纤维素的生物合成途径及其相关基因如蔗糖合成酶基因、KORRIG-AN基因等研究进展。     

Microfibrillar Structure, Cortical Microtubule Arrangement and the Effect of Amiprophos-methyl on Microfibril Orientation in the Thallus Cells of the Filamentous Green Alga, Chamaedoris orientalis

Microfibrillar structure, cortical microtubule orientation andthe effect of amiprophos-methyl (APM) on the arrangement ofthe most recently deposited cellulose microfibrils were investigatedin the marine filamentous green alga, Chamaedoris orientalis.The thallus cells of Chamaedoris showed typical tip growth.The orientation of microfibrils in the thick cell wall showedorderly change in longitudinal, transverse and oblique directionsin a polar dependent manner. Microtubules run parallel to thelongitudinally arranged microfibrils in the innermost layerof the wall but they are never parallel to either transverseor obliquely arranged microfibrils. The ordered change in microfibrilorientation is altered by the disruption of the microtubuleswith APM. The walls, deposited in the absence of the microtubules,showed typical helicoidal pattern. However, the original crossedpolylamellate pattern was restored by the removal of APM. Thissuggests that cortical microtubules in this alga do not controlthe direction of microfibril orientation but control the orderedchange of microfibril orientation. Amiprophos-methyl, Chamaedoris orientalis, coenocytic green alga, cortical microtubule, microfibrillar structure, tip growth     

Microfibril orientation across the secondary cell wall of Radiata pine tracheids

Polarised light microscopy and transmission electron microscopy were used to measure the variation in cellulose microfibril orientation across the secondary cell wall of Pinus radiata D. Don tracheids using oblique sections. Variations in orientation for the S1, S2 and S3 layers were determined for radial and tangential cell walls from the juvenile and mature earlywood at the base of the stem and at 5 m height. Microfibrils in the S1 layer are usually arranged in an S-helix (>90°) varying from 79° to 117° among tracheids. Within individual tracheids the microfibril orientation in the S1 region can be quite variable, sometimes changing from an S to Z-helix but without a well-defined crossed structure. Microfibrils in the S2 layer form a Z-helix (<90°) with average orientation varying from 1° to 59° among tracheids, while the S3 layer is also often a Z-helix varying from 50° to113° in orientation. The S2 layer shows significant variation among sample points within the tree related to age and height, while the S1 and S3 layers show small random variations within the tree. Microfibril orientation in the S2 layer is very uniform, although when variation does occur the trend is exclusively for an increase towards the S1 region. Orientation in the S3 layer often varies continuously from the S2 boundary to the lumen. The S3 layer becomes much thinner with increasing age and height, making measurement by light microscopy more difficult. The innermost lamellae of microfibrils at the lumen surface sometimes show a distinctive clustering, forming macrofibrils of varying orientation but this may be the result of delignification treatment.