Stage Of Secondary Wall Formation Research Articles

PtomtAPX is an autonomous lignification peroxidase during the earliest stage of secondary wall formation in Populus tomentosa Carr

At present, a cooperative process hypothesis is used to explain the supply of enzyme (class III peroxidases and/or laccases) and substrates during lignin polymerization. However, it remains elusive how xylem cells meet the needs of early lignin rapid polymerization during secondary cell wall formation. Here we provide evidence that a mitochondrial ascorbate peroxidase (PtomtAPX) is responsible for autonomous lignification during the earliest stage of secondary cell wall formation in Populus tomentosa. PtomtAPX was relocated to cell walls undergoing programmed cell death and catalysed lignin polymerization in vitro. Aberrant phenotypes were caused by altered PtomtAPX expression levels in P. tomentosa. These results reveal that PtomtAPX is crucial for catalysing lignin polymerization during the early stages of secondary cell wall formation and xylem development, and describe how xylem cells provide autonomous enzymes needed for lignin polymerization during rapid formation of the secondary cell wall by coupling with the programmed cell death process.

Characterization of xylan in the early stages of secondary cell wall formation in tobacco bright yellow-2 cells

The major polysaccharides present in the primary and secondary walls surrounding plant cells have been well characterized. However, our knowledge of the early stages of secondary wall formation is limited. To address this, cell walls were isolated from differentiating xylem vessel elements of tobacco bright yellow-2 (BY-2) cells induced by VASCULAR-RELATED NAC-DOMAIN7 (VND7). The walls of induced VND7-VP16-GR BY-2 cells consisted of cellulose, pectic polysaccharides, hemicelluloses, and lignin, and contained more xylan and cellulose compared with non-transformed BY-2 and uninduced VND7-VP16-GR BY-2 cells. A reducing end sequence of xylan containing rhamnose and galaturonic acid- residues is present in the walls of induced, uninduced, and non-transformed BY-2 cells. Glucuronic acid residues in xylan from walls of induced cells are O-methylated, while those of xylan in non-transformed BY-2 and uninduced cells are not. Our results show that xylan changes in chemical structure and amounts during the early stages of xylem differentiation.

Distribution of lignin and lignin precursors in differentiating xylem of Japanese cypress and poplar

Lignin is an integral component of the cell wall of vascular plants. The mechanism of supply of lignin precursors from the cytosol into the cell wall of differentiating xylem has not yet been elucidated. The present study showed that a certain amount of coniferyl alcohol glucoside (coniferin) occurred in the differentiating xylem of Japanese cypress (Chamaecyparis obtusa), as previously reported in gymnosperms. Coniferin content peaked in the early stages of secondary wall formation and decreased during lignification. In contrast to gymnosperms, coniferin content was limited in the differentiating xylem of poplar (Populus sieboldii × Populus grandidentata). Moreover, coniferyl alcohol was not detected in all specimens. In the differentiating xylem of poplar, a higher amount of sinapyl alcohol occurred than glucoside (syringin). However, the phloem contained syringin and not sinapyl alcohol. The sinapyl alcohol content in the xylem peaked in the cells with ceasing cell wall formation, and decreased gradually towards the boundary of the annual ring, where the lignin content kept increasing. Sinapyl alcohol in the differentiating xylem of poplar may be used for the lignification of the xylem.

Zeitliche Einordnung der G-Schicht-Auflagerung in den Prozess der Zellwandbildung bei Zugholzfasern in Zitterpappeln | On the chronology of g-layer formation during cell wall differentiation in tension wood fibres of poplars

The tension wood of some deciduous trees is characterised by fibres that form an additional cell wall layer, the so-called «gelatinous layer» (g-layer). The chronology of g-layer formation in the process of cell wall differentiation and lignification was investigated using two-year old poplars (Populus tremula L.). For this purpose the pinning-method was applied. The results show that the g-layer formation probably takes place at an early stage of secondary wall formation.

Xyloglucan endotransglycosylases have a function during the formation of secondary cell walls of vascular tissues.

Xyloglucan transglycosylases (XETs) have been implicated in many aspects of cell wall biosynthesis, but their function in vascular tissues, in general, and in the formation of secondary walls, in particular, is less well understood. Using an in situ XET activity assay in poplar stems, we have demonstrated XET activity in xylem and phloem fibers at the stage of secondary wall formation. Immunolocalization of fucosylated xylogucan with CCRC-M1 antibodies showed that levels of this species increased at the border between the primary and secondary wall layers at the time of secondary wall deposition. Furthermore, one of the most abundant XET isoforms in secondary vascular tissues (PttXET16A) was cloned and immunolocalized to fibers at the stage of secondary wall formation. Together, these data strongly suggest that XET has a previously unreported role in restructuring primary walls at the time when secondary wall layers are deposited, probably creating and reinforcing the connections between the primary and secondary wall layers. We also observed that xylogucan is incorporated at a high level in the inner layer of nacreous walls of mature sieve tube elements.

Relationship of growth cessation with the formation of diferulate cross-links and p-coumaroylated lignins in tall fescue leaf blades.

We examined relationships among cell wall feruloylation, diferulate cross-linking, p-coumarate deposition, and apoplastic peroxidase (EC 1.11.1.7) activity with changes in the elongation rate of leaf blades of slow and rapid elongating genotypes of tall fescue ( Festuca arundinacea Schreb.). Growth was not directly influenced by ferulic acid deposition but leaf elongation decelerated as 8-5-, 8- O-4-, 8-8-, and 5-5-coupled diferulic acids accumulated in cell walls. Growth rapidly slowed and stopped with the deposition of p-coumarate, which is primarily associated with lignification in grass cell walls. Accretion of ferulate, diferulates and p-coumarate continued after growth ended, into the later stages of secondary wall formation. The concentration of 8-coupled diferulates dwarfed that of the more commonly measured 5-5-coupled isomer, suggesting that the latter dimer is a poor indicator of diferulate cross-linking in cell walls. Further work is required to clearly demonstrate the role of diferulate cross-linking and p-coumaroylated lignins in the cessation of leaf growth in grasses.

Subcellular localization of expansin mRNA in xylem cells.

Terminal differentiation of many vascular cells involves cell wall changes. Cells first elongate their primary wall, then lay down a lignified secondary wall, which is often followed by digestion of the primary wall. Expansins are wall proteins that regulate wall changes, but little is known about the specific functions of the many individual expansin isoforms. An in vitro cell culture of synchronously differentiating tracheary elements was used to identify three new expansins and to compare their expression kinetics with the timing of wall changes. The genes encoding these expansins from zinnia (Zinnia elegans), designated ZeExp1, ZeExp2, ZeExp3, are expressed during cell elongation. ZeExp1 and ZeExp2 mRNA decrease at the early stage of secondary wall formation, whereas ZeExp3 does not. In planta, all three ZeExp mRNAs are found predominantly in a single flank of cells adjacent to protoxylem and metaxylem vessels and in cells roughly at the radial position of the fasicular and interfasicular cambium. Furthermore, within these cells, Exp mRNA is localized exclusively either to the apical or basipetal end of cells depending on the expansin gene and organ, providing the first evidence for polar localization of mRNA in plant cells. ZeExp1 and ZeExp3 mRNA are localized at the apical tip, whereas ZeExp2 mRNA is found in the basal tip. These observations indicate that these three expansins are xylem cell specific and possibly involved in the intrusive growth of the primary walls of differentiating xylem cells.

Cell Wall Biosynthesis during Silicification of Grass Hairs

Summary Cell walls of mature hairs from the lemma of Phalaris canariensis L. consisted of ≃ 40 % silica, 55% polysaccharides (containing 53% hexose, 45% pentose, and ⩽2% uronic acid residues) and ⩽ 5% protein. The ultrastructural form of the silica being deposited varied during development, the sequence being (1) sheet-like silica, (2) globular silica, (3) fibrillar silica. At the same time, a shift in the pattern of labelling of hair polysaccharides was detected in experiments where [ 14 C]glucose or [ 3 H]arabinose were fed to inflorescences of various ages for 32 h. At the early stages of secondary wall formation, when sheet-like silica was being deposited, arabinoxylan and cellulose were the major polymers being synthesised. Later, when globular silica was deposited, there was a dramatic increase in synthesis of «mixed-linked» β-(1→3, 1→4)- D -glucan relative to cellulose and arabinoxylan; some mannan was also labelled. In the final stages, during fibrillar silica deposition, little incorporation of sugars was occurring. It is argued that the changing composition of the organic phase of the cell wall may govern the size and ultrastructural arrangement of the silica particles being deposited.

Changes in the non-structural carbohydrate content of cotton (Gossypium spp.) fibres at different stages of development

The neutral sugars (glucose, fructose, and sucrose) and the sugar phosphates (glucose 6-phosphate, glucose 1-phosphate and fructose 6-phosphate) soluble in hot aqueous 80% methanol from the fibres of cotton - Gossypium arboreum L., G. barbadense L., and G. hirsutum L. - were determined at various stages of fibre development. In addition, the (1→3)-β-D-glucan content was measured and in the case of G. arboreum the rate of (1→3)-β-D-glucan and cellulose synthesis was determined with [(14)C]sucrose as the precursor. For each of the species a similar chronology was obtained for the changes in content of the various non-structural carbohydrates. At the early stages of secondary wall formation, glucose and fructose exhibited a maximum which was closely followed by a maximum in the (1→3)-β-D-glucan content and in the sugar phosphates. On the other hand, the sucrose content increased regularly until fibre maturity. The rates of synthesis of (1→3)-β-D-glucan and of cellulose were highest following the maximum in the (1→3)-β-D-glucan content, when the latter was being depleted.

(1→3)-β-D-Glucan (callose) is a probable intermediate in biosynthesis of cellulose of cotton fibres

The biosynthetic pathway of cellulose in higher plants is unknown1. Attempts to produce cellulose in vitro have shown that mainly (1→3)-β-D-glucan (callose) is produced from participate enzyme fractions of higher plants and uridine diphosphate glucose (UDPG)2–5. Some authors have claimed that in certain conditions, (1→4)-β-D-glucosidic linkages were also formed6–10. However, there is no conclusive evidence for the formation of pure (1→4)-β-D-glucans from UDPG. As caUose forms naturally not only in various parts of plants (sieve and pollen tubes, root hairs, cell plates) but also in response to wounding, it seems best to study cellulose biosynthesis in vivo rather than in wounded cells or cell homogenates which might produce callose but not cellulose. Using intact cotton fibres, we have now shown by pulse-chase experiments that callose has a high turnover and is likely to be an intermediate in cellulose biosynthesis at the stage of secondary wall formation.

Glucan synthesis by intact cotton fibres fed with different precursors at the stages of primary and secondary wall formation

Seed clusters of individual locules from fruit capsules of Gossypium arboreum L. with adhering intact fibres were fed with radioactive uridinediphosphoglucose (UDPG), guanosinediphosphoglucose (GDPG), glucose and sucrose. The incorporation into high molecular weight glucans of the fibres was studied. For primary wall fibres, UDPG at 1 mM was by far the best precursor, whereas sucrose was the best precursor for secondary wall fibres. No competition was observed between the incorporation of glucose from UDPG and from sucrose when the two were fed simultaneously to secondary wall fibres, indicating that their metabolic pathways are well separated when they are fed from the apoplast. Inhibitors of respiratory ATP-formation strongly inhibited incorporation of sucrose but not that of UDPG. Sucrose incorporation was studied at five different stages of development of the cotton fibres. At the stage of most intense secondary wall formation the incorporation rate was about 300 times that during primary wall formation (24 days post anthesis (DPA)). Incorporation from 1 mM UDPG or GDPG by secondary wall fibres (35 DPA) was less than twice that of primary wall fibres (22 DPA), indicating that the two sugar nucleotides are not readily used as precursors for secondary wall cellulose when they are fed to the exterior of intact cells. The high molecular weight non-cellulosic glucans formed from UDPG and sucrose at 5 and 1,000 μM were solubilized in strongly alkaline solutions or dimethyl-sulfoxide (DMSO) and were partially characterized by degradation with an exo-β-1,3-glucanase. After feeding for one hour, at most 1/3 of the radioactivity in high molecular weight material was found in cellulose and at least 2/3 in β-1,3-glucan. The proportions varied little for fibres in the age range of 30 to 48 DPA when sucrose was the precursor although the total incorporation varied by a factor of about four. The fact that at all stages of secondary wall formation β-1,3-glucan is synthesized at a very high rate, but that the total amount in the cell wall does not exceed 2% in the later stages of wall formation, can be interpreted in terms of a high turnover of this polysaccharide if it is assumed that wound effects are negligible in the system under study.

Hexagonally ordered ?rosettes? of particles in the plasma membrane ofMicrasterias denticulata Br�b. and their significance for microfibril formation and orientation

Cells ofMicrasterias denticulata Breb. at the stage of secondary wall formation have been studied by freeze-etching. It was found that the plasma membrane exhibits oval areas in which arrays of membrane particles occur. These particles form “rosettes” which are arranged in a hexagonally ordered lattice with a center to center spacing of ∼ 25 nm. Nearly the same periodicities can be found between microfibrils. It is concluded that the “rosettes” probably together with the thickened area of the plasma membrane below them represent the apparatus for the production and orientation of microfibrils. The hypothesis suggesting the incorporation of “membrane templates” functional in microfibril formation, originally advanced byKiermayer andDobberstein (1973) has received further support.

An examination of the developing cotton fiber: Wall and plasmalemma

The ultrastructure of developing cotton fibers has been examined using novel modifications of the techniques of surface replication, freeze-etching and thin-sectioning. The fiber surface was found to be coated with a lamellar cuticle, which is stretched and thinned as the fiber elongates. It is marked by bars which run parallel with the fiber long axis. Beneath the cuticle, the outermost microfibrils of the primary wall lie parallel with the fiber axis, while those adjacent to the plasma membrane are transverse. Primary wall microfibrils are present in bundles, disposed in left-handed and right-handed helices, which correspond with the fibrils observed optically. Microfibrils within bundles form in-phase waves, with wavelengths and amplitudes in the ranges 0.3–7 μm and 0.01–0.1 μm in primary and secondary walls respectively. As elongation proceeds bundles become displaced towards the cell axis. Microfibrils of the secondary wall, disposed around the cell as fast helices, are similarly bundled and wavy (though with a reduced amplitude). In surface-replicas, large (20–30 nm) granules are present on the cytoplasmic face of the wall which probably correspond with 20–40 nm low prominences visible on freeze-etch EF plasma membrane fracture faces. It is proposed that these may be microfibril-synthesizing centers. Plasma membranes fracture such that the membrane-associated-particles segregate 60∶40 between P and E fracture moieties, but the prominence and total number of these particles is reduced at the stage of secondary wall formation as compared with primary wall formation. Beneath the plasmalemma the axes of microtubules parallel secondary wall microfibril orientation. Cross-bridges, which stain heavily after glutaraldehyde/tannic acid fixation, link microtubules to plasma membrane. The use of butyl benzene to cement fragments of cotton fibers, employed in this work, may prove useful in other freeze-etch studies of long fibers which are readily ruptured during preparation.