A helicoidal cell wall texture in root hairs ofLimnobium stoloniferum

Summary The cell wall of root hairs ofLimnobium stoloniferum is composed of two fibrillar layers: an outer layer with a dispersed texture and an inner layer with a helicoidal texture. In stained oblique sections the helicoidal layer appears as a series of bow-shaped structures. In sections which were shadow-casted after the embedding medium was removed, the following properties of the helicoidal layer can be directly observed. (1) It is build up of superimposed lamellae. (2) Each lamella consists of parallel oriented microfibrils. (3) Going into the helicoidal layer, there is a counter-clockwise discontinuous rotation of the microfibril orientation in successive lamellae. (4) Between adjacent lamellae the average angular displacement of the microfibril orientation is about 23 degrees. The dispersed outer layer is also polylamellated, but with randomly arranged microfibrils in each lamella. Both layers are present in the lateral wall as well as in the apical wall of the root hairs. Observations indicate that in the cell wall of the tip the parallel oriented microfibrils of the outermost helicoidal lamellae become distorted towards a dispersed arrangement. The suggestion is made that the dispersed outer layer is derived from the helicoidal layer.     

Microtubules do not control microfibril orientation in a helicoidal cell wall

Summary The secondary cell wall layer of the young root hair ofEquisetum hyemale (L) has a helicoidal texture. The cortical microtubules in these hairs maintain an axial alignment while microfibrils are being deposited with a different orientation in each subsequent layer. The role of cortical microtubules in microfibril orientation is disputed.I gratefully acknowledge the support of Professor Dr. M. M. A.Sassen and the technical assistance of M.Wolters-Arts.     

Ultrastructure,differentiation and cell wall texture of trichoblasts and root hairs of ceratopteris thalictroides (L.) brongn. (Parkeriaceae)

Trichoblasts and root hairs of Ceratopteris thalictroides (L.) Brongn. were studied by different techniques to survey their morphological features. Trichoblasts could be identified at an early stage by an intra-vacuolar precipitate appearing during fixation. Special attention was paid to root-hair initiation. No structures or changes were observed that play a role in the initiation of papilla formation. When the papilla is formed, vesicles and periplasmic membranes can be observed which may play a role in the weakening of the cell wall during the papilla outgrowth.During root-hair growth, the nucleus of the trichoblast moves from the trichoblast to a subapical position in the root hair. The nuclei of all root cells contain 2 types of nuclear inclusions, one of which is proteinaceous.The cell wall of the Ceratopteris root hair has a helicoidal texture and because the cortical microtubules run longitudinally in the root hair, no correlation can be made between the directions of microtubules and microfibrils in these root hairs.     

Root hair-specific disruption of cellulose and xyloglucan in <Emphasis Type="Italic">AtCSLD3</Emphasis> mutants,and factors affecting the post-rupture resumption of mutant root hair growth

The glycosyl transferase encoded by the cellulose synthase-like gene CSLD3/KJK/RHD7 (At3g03050) is required for cell wall integrity during root hair formation in Arabidopsis thaliana but it remains unclear whether it contributes to the synthesis of cellulose or hemicellulose. We identified two new alleles, root hair-defective (rhd) 7-1 and rhd7-4, which affect the C-terminal end of the encoded protein. Like root hairs in the previously characterized kjk-2 putative null mutant, rhd7-1 and rhd7-4 hairs rupture before tip growth but, depending on the growth medium and temperature, hairs are able to survive rupture and initiate tip growth, indicating that these alleles retain some function. At 21°C, the rhd7 tip-growing root hairs continued to rupture but at 5oC, rupture was inhibited, resulting in long, wild type-like root hairs. At both temperatures, the expression of another root hair-specific CSLD gene, CSLD2, was increased in the rhd7-4 mutant but reduced in the kjk-2 mutant, suggesting that CSLD2 expression is CSLD3-dependent, and that CSLD2 could partially compensate for CSLD3 defects to prevent rupture at 5°C. Using a fluorescent brightener (FB 28) to detect cell wall (1 → 4)-β-glucans (primarily cellulose) and CCRC-M1 antibody to detect fucosylated xyloglucans revealed a patchy distribution of both in the mutant root hair cell walls. Cell wall thickness varied, and immunogold electron microscopy indicated that xyloglucan distribution was altered throughout the root hair cell walls. These cell wall defects indicate that CSLD3 is required for the normal organization of both cellulose and xyloglucan in root hair cell walls.     

Cortical microtubules and microfibril deposition in the cell wall of root hairs ofEquisetum hyemale

Summary The cell wall of root hairs ofEquisetum hyemale is shown to be composed of three different cell wall textures. The growing cell wall at the tip of the hair is composed of a dispersed texture of microfibrils, which continues along the outside of the whole hair. With increasing distance from the tip an increasing number of helicoidally arranged lamellae is deposited. These findings correspond with the observed isotropism of young hairs in polarized light.Hairs of approximately 4 days old become positive birefringent, indicating that longitudinally oriented layers prevail over layers with a transverse direction. This phenomenon starts at the base of the hair. Full-grown hairs are positive birefringent up to the tip and concordantly show a thick additional inner cell wall layer which forms a helical pattern the length of the hair, with a mean microfibril angle of 25 with the cell axis.Cortical microtubules, subjacent to the dispersed, the helicoidal and the helical wall texture are axially aligned and, thus, not in coalignment with the last deposited microfibrils.Coated and smooth vesicles are present in the cortical cytoplasm of both growing and full-grown hairs. Electron-dense profiles (20 nm in diameter), surrounded by a halo (of 50 nm) were observed on the wall-plasmalemma interface in full-grown hairs only. A relation of these structures with microfibril deposition could not be demonstrated. They might represent channels transporting material to the wall, which, in full-grown hairs, is heavily impregnated with a tawny brown substance.The general hypothesis that cortical microtubule orientation directs microfibril deposition is disputed.     

The helicoidal plant cell wall as a performing cellulose-based composite

The helicoidal plant cell wall can be considered as a composite in which cellulose is the constant reinforcing fiber. In order to strengthen the analogy with cholesteric liquid crystals, and taking into account a range of data, we describe a progressive series showing that cellulosic helicoidal systems are versatile and multifunctional. The following examples were considered: a) the cellulose microfibrils, with their rigid backbone possibly coated with a plastifying matrix; b) actual cholesteric cellulosic derivatives, such as in vitro liquid crystals and in vitro cellulosic mucilages; c) viscoplastic. growing cell walls; d) consolidated “stony” cell walls with their adaptation to intercellular communications. The series shows a dramatic progression from a liquid construction to what is the hardest in the plant cells, i.e. the sclerified walls.     

Nitrate induction of root hair density is mediated by TGA1/TGA4 and CPC transcription factors in Arabidopsis thaliana

Root hairs are specialized cells that are important for nutrient uptake. It is well established that nutrients such as phosphate have a great influence on root hair development in many plant species. Here we investigated the role of nitrate on root hair development at a physiological and molecular level. We showed that nitrate increases root hair density in Arabidopsis thaliana. We found that two different root hair defective mutants have significantly less nitrate than wild‐type plants, suggesting that in A. thaliana root hairs have an important role in the capacity to acquire nitrate. Nitrate reductase‐null mutants exhibited nitrate‐dependent root hair phenotypes comparable with wild‐type plants, indicating that nitrate is the signal that leads to increased formation of root hairs. We examined the role of two key regulators of root hair cell fate, CPC and WER, in response to nitrate treatments. Phenotypic analyses of these mutants showed that CPC is essential for nitrate‐induced responses of root hair development. Moreover, we showed that NRT1.1 and TGA1/TGA4 are required for pathways that induce root hair development by suppression of longitudinal elongation of trichoblast cells in response to nitrate treatments. Our results prompted a model where nitrate signaling via TGA1/TGA4 directly regulates the CPC root hair cell fate specification gene to increase formation of root hairs in A. thaliana.     

A new model for cellulose architecture in some plant cell walls

Summary A new model of rotating fibre components (helicoidal model) is proposed to explain the architecture of some plant cell walls. On the basis of tilting observations under the electron microscope, we establish the validity of this model for the cell wall ofChara vulgaris oospores. We suggest that this model explains the architecture seen in a number of published micrographs from a variety of different plant cell walls. Helicoidal architecture is shown to be distinct from the previously established crossed polylamellate architecture. The diagnostic features of helicoidal architecture are given. Morphogenesis of plant cell walls is discussed, with particular reference to self assembly in cholesteric liquid crystals.     

Time-course study and partial characterization of a protein on hyphae of arbuscular mycorrhizal fungi during active colonization of roots

Material on the surface of hyphal walls of arbuscular mycorrhizal fungi (AMF) during active colonization of plant roots was detected by a monoclonal antibody. Pot-cultured isolates of Glomus, Acaulospora, Gigaspora, Scutellospora, and Entrophospora had immunofluorescent material (IM) on younger, thinner, intact hyphae, but IM was scant to absent on thicker, melanized or lysing hyphae. Colonization of corn (Zea mays L.), Sudangrass (Sorghum sudanense (Piper) Staph.) or red clover (Trifolium pratense L.) was examined during 5 months of plant growth by removing cores and performing an indirect immunoassay on roots with attached hyphae. Fresh spores of some Glomus spp. had IM on the outer layer of the spore wall. Abundant IM was seen on root hairs of plants colonized by some isolates, and some IM was detected on root surfaces of all plants examined even during early colonization. After cultures were dried, hyphae, roots and spores had little to no IM. Uninoculated control roots had very rare, small patches of IM. An immunoreactive protein was extracted from hyphae of Gigaspora and Glomus isolates by using 20mM citrate (pH 7.0) at 121°C for 90 min. Gel electrophoresis profiles indicated that all isolates tested had the same banding patterns. Lectin-binding of extracted protein is suggestive of a glycoprotein. The immunofluorescence assay can be used to examine root sections for active colonization by AMF, and the potential use of the protein to quantify AMF activity in soil is discussed.     

Helicoidal orientation of cellulose microfibrils in Nitella opaca internode cells: ultrastructure and computed theoretical effects of strain reorientation during wall growth

The ultrastructure of the mature internode cell wall of Nitella opaca is described. It is interpreted in terms of a helicoidal array of cellulose microfibrils set in a matrix. A helicoid is a multiple plywood made up of layers of parallel microfibrils. There is a progressive change in direction from ply to ply, giving rise to characteristic arced patterns in oblique sections. A critical tilting test, using an electron microscope fitted with a goniometric stage, showed the expected reversal of direction of the arced pattern. Nitella cell wall is thus more regularly structured than previous studies have shown. From a survey of the cell-wall literature, we show that such arced patterns are common. This indicates that the helicoidal structure may be more widespread than is generally realised, although numerous other cell walls show no signs of it. Nevertheless, there are examples in most major plant taxa, and in several types of cells, including wood tracheids. Most of the examples, however, need confirmation by tilting evidence. There are possible implications for wall morphogenesis. Helicoidal cell walls might arise by selfassembly via a liquid crystalline phase, since it is known that the cholesteric state is itself helicoidal. A computer graphics programme has been developed to plot the expected effects of growth strain on the patterns in oblique sections of helicoids with various original angles between consecutive layers. Herringbone patterns typical of crossed polylamellate texture can be generated in this way, indicating a possible mode of their formation.     

The need for a constraining layer in the formation of monodomain helicoids in a wide range of biological structures

1. To be mechanically effective, supporting structures which are helicoidal need to be monodomain, with planar or concentric layers. 2. To achieve this in cholesteric liquid crystalline chemical models, a constraining surface is required. 3. The prediction which logically follows from this is that natural helicoidal systems in plant cell walls, spores, animal eggshells and cuticles need to be secreted within an initial constraining layer. 4. Evidence in support of this prediction is presented for a wide range of living systems, by reinterpretation of published work. This helps, at least partly, to explain the profusion of different kinds of layers in skeletal structures. 5. By contrast, systems lacking constraining layers have polydomain texture. 6. In plants, normal turgor pressure appears to be required for the deposition of monodomain helicoidal wall layers: reduced pressure leads to polydomain helicoid.     

Two separate zones of helicoidally orientated microfibrils are present in the walls ofNitella internodes during growth

Summary The dynamic changes in microfibril architecture in the internode cell walls of the giant unicellular algaNitella translucens were studied during cell expansion. Thin section electron microscopy in conjunction with mild matrix polysaccharide extraction techniques revealed three distinct architectural zones in the walls of fully grown cells. These zones were related to distinct phases of growth by monitoring changes in cell wall architecture of internodes during active cell expansion. The initial microfibril deposition before the onset of active cell growth is helicoidal. A helicoid is a structurally complex but ordered arrangement of microfibrils that has been detected increasingly often in higher plant cell walls. During active cell elongation microfibrils are deposited transversely to the direction of cell elongation as shown in earlier studies by birefringence measurements in the polarizing microscope. The gradual decline in cell elongation corresponds with a final helicoidal deposition which continues after cell expansion ceases entirely.The continual presence of the initial helicoidal zone in the outer wall region during the whole growth process suggests that these microfibrils do not experience strain reorientation and are continually reorganized, or maintained, in a well ordered helicoidal arrangement.     

Stimulation of root hair elongation in Arabidopsis thaliana by low phosphorus availability

Low phosphorus availability stimulates root hair elongation in many plants, which may have adaptive significance in soil phosphorus acquisition. We investigated the effect of low phosphorus on the elongation of Arabidopsis thaliana root hairs. Arabidopsis thaliana plants were grown in plant culture containing high (1000 mmol m^?3) or low (1 mmol m^?3) phosphorus concentrations, and root hair elongation was analysed by image analysis. After 15d of growth, low-phosphorus plants developed root hairs averaging 0.9 mm in length while high-phosphorus plants of the same age developed root hairs averaging 0.3 mm in length. Increased root hair length in low-phosphorus plants was a result of both increased growth duration and increased growth rate. Root hair length decreased logarithmically in response to increasing phosphorus concentration. Local changes in phosphorus availability influenced root hair growth regardless of the phosphorus status of the plant. Low phosphorus stimulated root hair elongation in the hairless axr2 mutant, exogenously applied IAA stimulated root hair elongation in wild-type high-phosphorus plants and the auxin antagonist CM PA inhibited root hair elongation in low-phosphorus plants. These results indicate that auxin may be involved in the low-phosphorus response in root hairs.     

Expansin gene loss is a common occurrence during adaptation to an aquatic environment

Expansins comprise a superfamily of plant cell wall loosening proteins that can be divided into four individual families (EXPA, EXPB, EXLA and EXLB). Aside from inferred roles in a variety of plant growth and developmental traits, little is known regarding the function of specific expansin clades, for which there are at least 16 in flowering plants (angiosperms); however, there is evidence to suggest that some expansins have cell‐specific functions, in root hair and pollen tube development, for example. Recently, two duckweed genomes have been sequenced (Spirodela polyrhiza strains 7498 and 9509), revealing significantly reduced superfamily sizes. We hypothesized that there would be a correlation between expansin loss and morphological reductions seen among highly adapted aquatic species. In order to provide an answer to this question, we characterized the expansin superfamilies of the greater duckweed Spirodela, the marine eelgrass Zostera marina and the bladderwort Utricularia gibba. We discovered rampant expansin gene and clade loss among the three, including a complete absence of the EXLB family and EXPA‐VII. The most convincing correlation between morphological reduction and expansin loss was seen for Utricularia and Spirodela, which both lack root hairs and the root hair expansin clade EXPA‐X. Contrary to the pattern observed in other species, four Utricularia expansins failed to branch within any clade, suggesting that they may be the result of neofunctionalization. Last, an expansin clade previously discovered only in eudicots was identified in Spirodela, allowing us to conclude that the last common ancestor of monocots and eudicots contained a minimum of 17 expansins.     

Helicoidal pattern in secondary cell walls and possible role of xylans in their construction

The helicoidal organization of secondary cell walls is overviewed from several examples. Both the plywood texture and the occurrence of characteristic defects strongly suggest that the wall ordering is relevant of a cholesteric liquid-crystal assembly that is rapidly and strongly consolidated by lignification. A preferential localization of glucuronoxylans, major matrix components, and in vitro re-association experiments emphasize their preeminent role: (1) during the construction of the composite as directing the cellulose microfibrils in a helicoidal array; (2) during the lignification of the composite as a host structure for lignin precursors.     

慈姑根尖细胞微管骨架的免疫荧光观察

分别用考马斯亮蓝染色和间接免疫荧光标记,并运用荧光倒置显微镜和激光共聚焦显微镜,对慈姑根尖固定后酶解获得的去壁细胞和细胞团块以及根尖细胞分裂周期中微管骨架列阵进行详细观察,以探索高等植物微管周期的普遍性。结果表明:慈姑根尖固定后酶解可获得大量结构完整的去壁细胞与细胞团块;考马斯亮蓝染色观察可见,慈姑根尖细胞中丰富的蛋白物质以及处于不同分裂期的细胞核染色体;免疫荧光观察可见,慈姑根尖细胞周期中微管骨架保存较好,主要有周质微管、早前期带微管、纺缍体微管和成膜体微管4种循序变化的排列方式,构成了高等水生植物分裂细胞中典型的微管周期。实验结果证明,高等水生植物与陆生植物微管周期具有相似性,为植物微管周期概念提供了新的实例。     

Mechanisms of infection of plants by nitrogen fixing organisms

Heterotrophic nitrogen-fixing microorganisms can enter plants via wounds, root hairs or intact epidermises. All at some stage need the ability to digest primary cell walls and/or middle lamellas. None appears to digest secondary walls. The ability of any organism to infect a particular plant reflects (a) the enzymes produced by the microorganism (and possibly, as part of its reaction, the plant); (b) the exact nature of the primary wall; (c) the distribution of secondary walls. Plants may respond to infection by hypersensitive and other reactions which could be triggered by production of cell wall fragments. Infection threads of secondary wall material may be essential for root hair infection and where cell boundaries are crossed. Entry into host cells other than by infection threads involves a delicate balance between endophyte and host. This may only be achieved in one or a few cells, which may then divide repeatedly to produce a symbiotic structure.     

Pectic homogalacturonan masks abundant sets of xyloglucan epitopes in plant cell walls

Background  

Molecular probes are required to detect cell wall polymers in-situ to aid understanding of their cell biology and several studies have shown that cell wall epitopes have restricted occurrences across sections of plant organs indicating that cell wall structure is highly developmentally regulated. Xyloglucan is the major hemicellulose or cross-linking glycan of the primary cell walls of dicotyledons although little is known of its occurrence or functions in relation to cell development and cell wall microstructure.