Distribution of alginate and cellulose and regulatory role of calcium in the cell wall of the brown alga <Emphasis Type="Italic">Ectocarpus siliculosus</Emphasis> (Ectocarpales,Phaeophyceae)

Main conclusion

This work investigated a correlation between the three-dimensional architecture and compound–components of the brown algal cell wall. Calcium greatly contributes to the cell wall integrity.Brown algae have a unique cell wall consisting of alginate, cellulose, and sulfated polysaccharides. However, the relationship between the architecture and the composition of the cell wall is poorly understood. Here, we investigated the architecture of the cell wall and the effect of extracellular calcium in the sporophyte and gametophyte of the model brown alga, Ectocarpus siliculosus (Dillwyn) Lyngbye, using transmission electron microscopy, histochemical, and immunohistochemical studies. The lateral cell wall of vegetative cells of the sporophyte thalli had multilayered architecture containing electron-dense and negatively stained fibrils. Electron tomographic analysis showed that the amount of the electron-dense fibrils and the junctions was different between inner and outer layers, and between the perpendicular and tangential directions of the cell wall. By immersing the gametophyte thalli in the low-calcium (one-eighth of the normal concentration) artificial seawater medium, the fibrous layers of the lateral cell wall of vegetative cells became swollen. Destruction of cell wall integrity was also induced by the addition of sorbitol. The results demonstrated that electron-dense fibrils were composed of alginate-calcium fibrous gels, and electron negatively stained fibrils were crystalline cellulose microfibrils. It was concluded that the spatial arrangement of electron-dense fibrils was different between the layers and between the directions of the cell wall, and calcium was necessary for maintaining the fibrous layers in the cell wall. This study provides insights into the design principle of the brown algal cell wall.

     

Ecology and evolution of the diaspore "burial syndrome"

Hygroscopically active awns or "bristles" have long intrigued scientists. Experimental evidence shows that they are important for diaspore burial in the correct orientation, thereby increasing successful seed germination and seedling survival. Despite these ecological advantages, 38 of the 280 species of grasses in Danthonioideae lack awns. We provide the first study of awns in a phylogenetic context and show that although the awnless state has arisen ca. 25 times independently, the ecological disadvantage of not having an awn also applies in an evolutionary context. Only in Tribolium and Schismus have awnless ancestors diversified to form a clade of primarily awnless descendents. Several of the awnless species in these genera are annual and we find a significant correlation between the evolution of awns and the evolution of life history. A suite of other diaspore traits accompany the awned or awnless states. We interpret the awn as being the visible constituent of a compound "burial syndrome," the two ecological extremes of which may explain the correlation between awns and life history and provide an explanation why awnless species in Tribolium and Schismus persist.     

An-1 Encodes a Basic Helix-Loop-Helix Protein That Regulates Awn Development,Grain Size,and Grain Number in Rice

Long awns are important for seed dispersal in wild rice (Oryza rufipogon), but are absent in cultivated rice (Oryza sativa). The genetic mechanism involved in loss-of-awn in cultivated rice remains unknown. We report here the molecular cloning of a major quantitative trait locus, An-1, which regulates long awn formation in O. rufipogon. An-1 encodes a basic helix-loop-helix protein, which regulates cell division. The nearly-isogenic line (NIL-An-1) carrying a wild allele An-1 in the genetic background of the awnless indica Guangluai4 produces long awns and longer grains, but significantly fewer grains per panicle compared with Guangluai4. Transgenic studies confirmed that An-1 positively regulates awn elongation, but negatively regulates grain number per panicle. Genetic variations in the An-1 locus were found to be associated with awn loss in cultivated rice. Population genetic analysis of wild and cultivated rice showed a significant reduction in nucleotide diversity of the An-1 locus in rice cultivars, suggesting that the An-1 locus was a major target for artificial selection. Thus, we propose that awn loss was favored and strongly selected by humans, as genetic variations at the An-1 locus that cause awn loss would increase grain numbers and subsequently improve grain yield in cultivated rice.     

Stomatal control of gas exchange in barley awns

The gas exchange of barley ears and awns was measured in the field using a gas analysis system and a diffusion porometer. Awn stomatal resistance decreased with increasing irradiance but to a smaller extent than leaf stomatal resistance. Measurements on ears immediately before and after successively removing awns showed that awn transpiration and photosynthesis were proportional to awn area and that awns accounted for 73% of transpiration by the ear. Although the maximum rates of photosynthesis of which awns were capable declined with age, awns accounted for 80–115% of the net CO₂ uptake of complete ears because the ears-less-awns could respire more CO₂ than they absorbed. Ear photosynthesis accounted for 52% of the weekly increment in ear dry weight after ear emergence, but 5 weeks later photosynthesis by the ear balanced respiration. Overall photosynthesis by the ear accounted for 35 % of its final weight. Differences in the light response curves of leaves and ears can be fully accounted for by the different relationships between stomatal resistance and irradiance of the two organs.     

Wall fibrils ingerminating sporangiospores of Gilbertella persicaria (Mucorales)

Summary In germinated sporangiospores of Gilbertella persicaria, negatively contrasted fibrils, 20–70 Å diam, are seen in thin sections of the inner vegetative wall that is continuous with the germ tube wall. The fibrils are randomly oriented in a loose network in this wall and in the germ tube wall. Germ tubes have an additional surface layer of fine, positively contrasted fibrils which appear as a nap-like coating on the hyphae. Patterns of wall fibril orientation are not revealed by transverse sections of spore and germ tube walls, whereas oblique and tangential sections are favorable for examining cell wall architecture in situ. Staining patterns show textural and compositional differences among various wall layers.     

THE CELL WALL OF COSMARIUM BOTRYTIS12

The cell wall of Cosmarium botrytis was studied through the use of the freeze-etch technique. The cell wall consists of many thin layers. Fracturing along one layer reveals the positioning of the wall sculpturing, wall pores, and wall microfibrils. The individual microfibrils are grouped together in bands of parallel oriented fibrils. The different bands of parallel microfibrils were apparently arranged at random angles with regard to each other. Small particles may also be present in the cell walls. The cell wall pore unit of Cosmarium botrytis was studied through the use of scanning, freeze-etching, and thin sectioning techniques. The pore sheaths, on the outside of the cell wall, form a collar around the mouth of each pore. The pore sheath is composed of needle-like fibrils radiating outward from the pore. A pore channel traverses the cell wall and leads to a complex pore bulb region between the cell wall and the plasmalemma. The pore bulb contains many small fibrils which radiate toward the plasmalemma from a number of net-like fibril layers which in turn merge into a very electron dense region near the base of the pore.     

A barrier covering lignified cell walls of barley straw that restricts access by rumen microorganisms

In barley straw, all stem tissues except the chlorenchyma are lignified. Electronmicroscopic investigations employing staining for cell-wall constituents and replica techniques revealed the presence of a tertiary wall covering the secondary wall and a warty layer deposited on the tertiary wall. The tertiary wall was composed of cellulose fibrils orientated in all directions, which were embedded in matrix material. The warty layer was comprised of granules and globules that were in many instances associated with a thin, flat, continuous layer. Though the tertiary wall could be degraded, the warty layer was resistant to degradation by rumen microorganisms. Only where the warty layer was mechanically disrupted could underlying cell-wall material be degraded. Bacteria could also burrow beneath these layers from exposed cell walls at the cut edges of the plant material. The warty layer forms a barrier to cell-wall-degrading bacteria and limits their colonization of straw stem.     

Cell locomotion forces versus cell contraction forces for collagen lattice contraction: an in vitro model of wound contraction

Cultured human dermal fibroblasts suspended in a rapidly polymerizing collagen matrix produce a fibroblast-populated collagen lattice. With time, this lattice will undergo a reduction in size referred to as lattice contraction. During this process, two distinct cell populations develop. At the periphery of the lattice, highly oriented sheets of cells, morphologically identifiable as myofibroblasts, show cell-to-cell contacts and thick, actin-rich staining cytoplasmic stress fibers. It is proposed that these cells undergoing cell contraction produce a multicellular contractile unit which reorients the collagen fibrils associated with them. The cells in the central region, referred to as fibroblasts, are randomly oriented, with few cell-to-cell contacts and faintly staining actin cytoplasmic filaments. In contrast it is proposed that cells working as single units use cell locomotion forces to reorient the collagen fibrils associated with them. Using this model, we sought to determine which of these two mechanisms, cell contraction or cell locomotion, is responsible for the force that contracts collagen lattices. Our experiments showed that fibroblasts produce this contractile force, and that the mechanism for lattice contraction appears to be related to cell locomotion. This is in contrast to a myofibroblast; where the mechanism for contraction is based upon cell contractions. Fibroblasts attempting to move within the collagen matrix reorganize the surrounding collagen fibrils; when these collagen fibrils can be organized no further and cell-to-cell contacts develop, which occurs at the periphery of the lattice first, these cells can no longer participate in the dynamic aspects of lattice contraction.     

Cellulose orientation at the surface of the Arabidopsis seedling. Implications for the biomechanics in plant development

In diffuse growing cells the orientation of cellulose fibrils determines mechanical anisotropy in the cell wall and hence also the direction of plant and organ growth. This paper reports on the mean or net orientation of cellulose fibrils in the outer epidermal wall of the whole Arabidopsis plant. This outer epidermal wall is considered as the growth-limiting boundary between plant and environment. In the root a net transverse orientation of the cellulose fibrils occurs in the elongation zone, while net random and longitudinal orientations are found in subsequent older parts of the differentiation zone. The position and the size of the transverse zone is related with root growth rate. In the shoot the net orientation of cellulose fibrils is transverse in the elongating apical part of the hypocotyl, and longitudinal in the fully elongated basal part. Leaf primordia and very young leaves have a transverse orientation. Throughout further development the leaf epidermis builds a very complex pattern of cells with a random orientation and cells with a transverse or a longitudinal orientation of the cellulose fibrils. The patterns of net cellulose orientation correlate well with the cylindrical growth of roots and shoots and with the typical planar growth of the leaf blade. On both the shoot and the root surface very specific patterns of cellulose orientation occur at sites of specific cell differentiation: trichome-socket cells complexes on the shoot and root hairs on the root.     

Structure and Photosynthetic Characteristics of Awns of Wheat and Barley

The surface and the cross section of awns of wheat and barley were examined by scanning electron microscopy,ultrastructure of cells were observed under a transmisson electron microscope and the photosynthetic rates were measured with an oxygen, electrode and infra-red CO2 analyser. The main results were as follows :The cross section of wheat awn appeared to be acutely trianglular whereas that of barley awn was obtusely triangular. There were rows of stomota on either side of epidermis in both wheat and barley awns. Under the stomatic band there were green tissues. The green cells in the awn were differentiated from the parenchyma cells . The mature green cells possessed papillae which were rich in chloroplasts and mitochondria. The tamella system in chloroplasts was well developed and contained many starch grains. There were three vascular bundles in each awn. The sheath cells near the green tissues contained chloroplasts. The photosynthate in the green cells might pass through the sheath cells and companion cells to sieve elements. The highest photosynthetic rate of the awn was seen at the flowering stage ,reaching about 20 μmol CO2·m-2·s-1. The light compensation point was 70—80 μE·m-2· s-1. The light saturation point was about 1500 μE·m-2·s-1. The CO2 compensation point was 50—60 ppm and the CO2 saturation point was about 900ppm . The photosynthetic rate and stomatal conductance were easily effected by CO2 concentration, light intensity and the duration of illumination . There was a positive correlation between the photosynthetic rate and the chloro-phyll content in the awns. The CO2-releasing rate in photorespiration of awn was about 4–5 μmol CO2·m-2·s-1.     

Out-of-plane orientation of cellulose elementary fibrils on spruce tracheid wall based on imaging with high-resolution transmission electron microscopy

Main conclusion

A 3D model of the tracheid wall has been proposed based on high-resolution cryo-TEM where, in contrast to the current understanding, the cellulose elementary fibrils protrude from the cell wall plane. The ultrastructure of the tracheid walls of Picea abies was examined through imaging of ultrathin radial, tangential and transverse sections of wood by transmission electron microscopy and with digital image processing. It was found that the elementary fibrils (EFs) of cellulose were rarely deposited in the plane of the concentric cell wall layers, in contrast to the current understanding. In addition to the adopted concept of longitudinal fibril angle, EFs protruded from the cell wall plane in varying angles depending on the layer. Moreover, the out-of-plane fibril angle varied between radial and tangential walls. In the tangential S₂ layers, EFs were always out-of-plane whereas planar orientation was typical for the S₂ layer in radial walls. The pattern of protruding EFs was evident in almost all axial and transverse images of the S₁ layer. Similar out-of-plane orientation was found in the transverse sections of the S₃ layer. A new model of the tracheid wall with EF orientation is presented as a summary of this study. The outcome of this study will enhance our understanding of the elementary fibril orientation in the tracheid wall.     

Distribution of carbon-14 assimilated by wheat awns

The pattern of distribution of carbon assimilated by awns was investigated in two lines of Triticum aestivum. Single awns on basal florets of spikelets in the central part of the ear were dosed with ¹⁴CO₂. Five days after dosing, 99% of the carbon-14 recovered was in the spikelet bearing the awn. Of the carbon-14 exported from the treated awn 57% went to the grain of the first floret, 1% to the second, 28% to the third and 7% to the fourth.     

Taxonomic separation ofKengyilia (Poaceae: Triticeae) in relation to nearest relatedRoegneria,Elymus, andAgropyron,based on some morphological characters

Based on 100 species representative of the four genera, we scored 290 herbarium specimens for a number of morphological characters. The data were subjected to canonical discriminant analysis using characters different from those in the identification key to these genera byBaum, Yen, andYang (1991). These characters collectively support the four groupsAgropyron, Kengyilia, Roegneria, andElymus as previously defined. The four groups are also supported by the linear discriminant function with an overall rate of 83% correct classification. Length of lemma awn was found to be an additional diagnostic character asAgropyron andKengyilia have lemma awns shorter than 5 mm, whereasRoegneria andElymus have longer lemma awns with very few exceptions. Length of glume awns is also a useful supplementary generic diagnostic.Agropyron andElymus have glume awns, whereas the majority of species ofKengyilia and more than half of the species ofRoegneria lack them. If a glume awn is present it is usually not longer than 1 mm.     

A ZYGOTE-SPECIFIC PROTEIN WITH HYDROXYPROLINE-RICH GLYCOPROTEIN DOMAINS AND LECTIN-LIKE DOMAINS INVOLVED IN THE ASSEMBLY OF THE CELL WALL OF CHLAMYDOMONAS REINHARDTII (CHLOROPHYTA)

The cell wall of Chlamydomonas reinhardtii zygotes, which forms rapidly after the fusion of wall-free gametes, provides a tractable system for studying the properties and assembly of hydroxyproline-rich glycoproteins, the major proteinaceous components of green algal and plant cell walls. We report the cloning of the zsp2 gene and the analysis of its ZSP-2 product, a 58.9 kDa polypeptide that is synthesized exclusively by zygotes. The protein contains two (SP)_x repeats, establishing it as a member of the cell wall hydroxyproline-rich glycoproteins family. It also contains a 4-fold iteration of an amino acid sequence centered around cysteine residues, a configuration found in both plant and animal lectins. Furthermore, we report four observations on pellicle composition and production. First, cell-free preparations of the pellicle matrix are rich in hydroxyproline, arabinose, and galactose and contain bundles of very long fibrils. Second, glutathione blocks pellicle formation and results in the accumulation of long fibrils in the growth medium. Third, antibody to ZSP-2 also blocks pellicle formation. Fourth, ZSP-2 immunolocalizes to the boundary between the outer layers of the wall proper and the pellicle matrix. These observations are consistent with the possibility that the Cys-rich (glutathione-sensitive) lectin-like domains of ZSP-2 may bind to sugar residues on the long fibrils and anchor them to the cell wall, thereby initiating and maintaining pellicle formation.     

Microstructure and crystallographic-texture of giant barnacle (Austromegabalanus psittacus) shell

Barnacle shell is a very complex and strong composite bioceramic composed of different structural units which consist of calcite 15 microcrystals of very uniform size. In the study reported herein, the microstructural organization of these units has been examinated in detail with optical and scanning electron microscopy, and X-ray diffraction techniques. These analyses showed that the external part of the shell has a massive microstructure consisting of randomly oriented crystals. Toward the interior, the shell became organized in mineral layers separated by thin organic sheets. Each of these mineral layers has a massive microstructure constituted by highly oriented calcite microcrystals with their c-axes aligned [(001) fibre texture] perpendicular to the organic sheets and the shell surface. Interestingly, in another structural unit, the shell shield, the orientation of the c-axis calcite crystals shifts from being perpendicular to being parallel to the shell surface across its thickness. This study provides evidence that the organic matrix is responsible for the organization of the shell mineral and exterts strong a strict control on the polymorphic type, size and orientation of shell-forming crystals.     

Cell expansion in the epidermis: microtubules, cellulose orientation and wall loosening enzymes

Two models of isolated epidermis were used to demonstrate that the net orientation of cellulose microfibrils in the cell wall is related to mechanical properties of the tissue, and can be used as an indicator for wall anisotropy. In the developing plant epidermis, cells expand in one or two directions in the plane of the plant surface. In epidermis cells actively expanding in one direction (elongation), the orientation of cortical microtubules closely matches the net cellulose orientation. In epidermis cells expanding in two directions, the orientation of the parallel microtubules does not coincide with the net cellulose orientation in the adjacent cell wall. The orientation of cortical microtubules is thus not always a reliable indicator of wall characteristics. In both types of epidermis, a high rate of expansion correlates with a high activity of xyloglucan endotransglycosylase (XET), as determinedin situ. This high activity alone cannot explain unidirectional wall expansion.     

Heteropogon-Themeda grasses evolve to occupy either tropical grassland or wetland biomes

Species of the Heteropogon-Themeda clade are ecologically important grasses distributed across the tropics, including widespread species, such as the pantropical Heteropogon contortus and Themeda triandra, and range-restricted species such as Heteropogon ritchiei and Themeda anathera. Here, we examine habitat preferences of the grassland/savanna and wetland species by describing bioclimatic niche characteristics, characterizing functional traits, and investigating the evolution of functional traits of 31 species in the Heteropogon-Themeda clade in relation to precipitation and temperature. The climatic limits of the clade are linked to mean annual precipitation and seasonality that also distinguish seven wetland species from 24 grassland/savanna species. Tests of niche equivalency highlighted the unique bioclimatic niche of the wetland species. However, climatic factors do not fully explain species geographic range, and other factors are likely to contribute to their distribution ranges. Trait analyses demonstrated that the wetland and grassland/savanna species were separated by culm height, leaf length, leaf area, awn length, and awn types. Phylogenetic analyses showed that the wetland species had tall stature with long and large leaves and lack of hygroscopic awns, which suggest selective pressures in the shift between savanna/grassland and wetland. The two most widespread species, H. contortus and T. triandra, have significantly different bioclimatic niches, but we also found that climatic niche alone does not explain the current geographic distributions of H. contortus and T. triandra. Our study provides a new understanding of the biogeography and evolutionary history of an ecologically important clade of C₄ tropical grasses.     

FT-IR study of the Chara corallina cell wall under deformation

Fourier-transform infrared (FT-IR) microspectroscopy was used to investigate both the chemical composition of, and the effects of an applied strain on, the structure of the Chara corallina cell wall. The inner layers of the cell wall are known to have a transverse cellulose orientation with a gradient through the thickness to longitudinal orientation in the older layers. In both the native state and following the removal of various biopolymers by a sequential extraction infrared dichroism was used to examine the orientation of different biopolymers in cell-wall samples subjected to longitudinal strain. In the Chara system, cellulose microfibrils were found to be aligned predominantly transverse to the long axis of the cell and became orientated increasingly transversely as longitudinal strain increased. Simultaneously, the pectic polysaccharide matrix underwent molecular orientation parallel to the direction of strain. Following extraction in CDTA, microfibrils were orientated transversely to the strain direction, and again the degree of transverse orientation increased with increasing strain. However, the pectic polysaccharides of the matrix were not detected in the dichroic difference spectra. After a full sequential extraction, the cellulose microfibrils, now with greatly reduced crystallinity, were detected in a longitudinal direction and they became orientated increasingly parallel to the direction of strain as it increased.