Primary walls

Structure of the Cell Wall. The iaterior stmcture of the ceU wall is shown in Figure 6. The interfiber region is the middle lamella (ML). This region, rich in lignin, is amorphous and shows no fibnUar stmcture when examined under the electron microscope. The cell wall is composed of stmcturaHy different layers or lamellae, reflecting the manner in which the cell forms. The newly formed cell contains protoplasm, from which cellulose and the other cell wall polymers are laid down to thicken the cell wall internally. Thus, there is a primary wall (P) and a secondary wall (S). The secondary wall is subdivided into three portions, the S, S2, and layers, which form sequentially toward the lumen. Viewed from the lumen, the cell wall frequendy has a bumpy appearance. This is called the warty layer and is composed of protoplasmic debris. The warty layer and exposed layer are sometimes referred to as the tertiary wad. 

Cotton is essentially 95% cellulose [9004-34-6] (Table 1). The nonceUulosic materials, consisting mostly of waxes, pectinaceous substances, and nitrogenous matter, are located to a large extent in the primary wall, with small amounts in the lumen (23). 

All secondary cell walls develop from primary cell walls. Cells no longer grow once lignin is added to their wails. Lignification, which is a key step in the conversion of a primary cell wall into a secondary cell wall, results in terminal differentiation of the encased cell. Indeed, many cells with lignified walls die. The totipotency of plant cells is limited to cells enveloped in primary walls. 

The pectin network.-The second polysaccharide network present in primary cell walls is composed of pectic polysaccharides. The pectin network appears to coexist with the cellulose/hemicellulose network, that is, both networks appear to be able to share the same space [16-19]. However, the proportions of the two networks appear to vary from location to location within a single cell wall as well as from the primary wall of one type of cell to the primary wall of a another type of cell [9,20-22]. 

The pectin network revisited.--The importance of the interconnections of the pectic polysaccharides to the integrity of the pectin network has been highlighted by the recent discovery that RG-II is present in primary walls as a mixture of monomers and dimers [54]. The dimers are covalently cross-linked by borate diesters [55,56]. If single molecules of homogalacturonan are covalently attached to both RG-I and RG-II, the covalently cross-linked RG-n dimers would explain how the network of the three types of pectic polysaccharides is covalently connected and covalently cross-linked. 

This essay was written in an attempt to explain our overview of primary cell walls and to reach consensus on the nomenclature of primary cell wall polysaccharides. We present evidence supporting the hypothesis that cellulose, xyloglucan, arabinoxylan, homogalacturonan, RG-I, and RG-II are the six polysaccharides common to all primary cell walls of higher plants. In many cells, these six polysaccharides account for all or nearly all of the primary wall polysaccharides. Like the physically interacting proteins that constitute the electron transport machinery of mitochondria, the structures of the six patently ubiquitous polysaccharides of primary cell walls have been conserved during evolution. Indeed, we hypothesize that the common set of six structural polysaccharides of primary cell walls have been structurally... 

Our two network model of the primary wall receives support from a variety of indirect observations. For example it has been shown Aat when a cell wall is regenerated by a carrot protoplast a homogalacturonan/ rhamnogalacturonan shell is laid down first, through which the cellulose/ hemicellulose network is later intercalated (8). Further evidence that pectin may form an independent network is seen in the fact that walls from suspension-cultured cells of tofnato Lycopersicon esculentum VF 36),... 

The primary walls of growing plant cells are composed of 90% carbohydrate and 10% protein (51). Carbohydrate in the primary wall is present predominantly as cellulose, hemicellulose, and pectin. The pectic polysaccharides, are defined as a group of cell wall polymers containing a-l,4-linked D-galactosyluronic acid residues (62,76). Pectic polysaccharides are a major component of the primary cell waU of dicots (22-35%), arc abundant in gymnosperms and non-graminaceous monocots, and are present in reduced amounts (-10%) in the primary walls of the graminaceae (27,62). 

Immunogold localization of the pectic epitope has been performed on different types of cells cell suspensions, roots, shoots, meristems, coleoptiles, pollen grains, protoplasts from different species carrot, sugar beet, tobacco, oat... The pattern of labeling was always the same polygalacturonic acid was essentially located on the material expanded at three-way junctions between cells or lining intercellular space, but was not found in primary walls. No epitope could be located close to the plasma membrane (Fig. lO.a). Middle lamellae far from junction zones and walls of meristematic cells were never labeled. 

However, since pectins can be methylesterified and/or acetylesterified, sections were treated on grid with an orange peel methylesterase to remove the methyl groups or with NaOH to remove both methyl and acetyl groups. After the enzymatic treatment, all the primary walls of most of the samples bind the... 

Fig. 10. Intercellular junction zones of carrot cells grown in suspension have been observed in electron microscopy after immunogold labeling with the 2F4 antibody, (a) no treatment of the sections prior to labeling the gold particles are restricted to the center of the junction zones (b) enzymatic (pectin methyl esterase) deesterification of the E.M. grids before labeling the deesterified pectins present in the primary walls now bind the probe. Scale bars = 1 pm.

The components of the plant cell wall (8-21) are the middle lamella (intercellular substance), the primary wall, and the secondary wall. The middle lamella is the pectic layer between cells and holds adjoining cells together as do membrane carbohydrates. The primary wall is thin (1-3 pm) and flexible containing cellulose, hemicelluloses, pectins, and glycoproteins. This wall provides mechanical strength, maintains cell shape,... 

The secondary wall is located inside the primary wall and can possess pits and sometimes three distinct layers, Sl5 S2, and S3. The secondary wall can contain 25% lignin, and cellulose is more abundant than in primary walls. Support and resistance to decay are the main functions of the primary wall. 

The outer secondary cell wall (SI) is comparable in thickness to the primary wall and consists of four to six lamellae which spiral in opposite directions around the longitudinal axis of the tracheid. The main bulk of the secondary wall is contained in the middle secondary cell wall (S2), and may be as little as 1 fim thick in early woods and up to 5 fim in summer wood. The microfibrils of this part of the wall spiral steeply about the axial direction at an angle of around 10 to 20°. The inner secondary wall (S3), sometimes also known as the tertiary wall, is not always well developed, and is of no great technological importance. 

A clear understanding of lignin deposition in the cell wall is not yet possible, but a number of facts are known. Lignin precursors of the phenylglucoside type are formed either in the region of the cambium (the zone of new cell synthesis) or within the lignifying cell itself. Lignification is thus initiated in the differentiated wood cells from the primary walls adjacent to the cell corners and then extends into the inter-cellular area, the lamella, and thereafter to the primary and secondary cell walls. 

Cellulose microfibrils make up the basic framework of the primary wall of young plant cells (3), where they form a complex network with other polysaccharides. The linking polysaccharides include hemicellulose, which is a mixture of predominantly neutral heterogly-cans (xylans, xyloglucans, arabinogalactans, etc.). Hemicellulose associates with the cellulose fibrils via noncovalent interactions. These complexes are connected by neutral and acidic pectins, which typically contain galac-turonic acid. Finally, a collagen-related protein, extensin, is also involved in the formation of primary walls. 

The major polymers that make up the wall are polysaccharides and lignin. These occur together with more minor but very important constituents such as protein and lipid. Water constitutes a major and very important material of young, primary walls (2). The lignin is transported in the form of its building units (these may be present as glucosides) and is polymerized within the wall. Those polysaccharides which make up the matrix of the wall (hemicelluloses and pectin material) are polymerized in the endomembrane system and are secreted in a preformed condition to the outside of the cell. Further modifications of the polysaccharides (such as acetylation) may occur within the wall after deposition. Cellulose is polymerized at the cell surface by a complex enzyme system transported to the plasma membrane (3). 

Figure 4. The composition of radioactivity in neutral sugars. Arabic numerals are the fraction numbers. The differentiating stages in each fraction are as follows Fractions 1-2, primary wall stage 3, Si stage 4-6, S2 stage 7-8, S3 stage.

Several reticula of the r-ER show an ordered arrangement and many ribosomes are attached to their membrane during the primary wall development stage. As maturation proceeds, the r-ER s then gradually decrease not only in number and length of reticula, but also in the number of ribosomes. The s-ER s, on the other hand, become largest after the S3 stage, and sometimes attach ribosomes at their terminals. 

During primary wall formation the plastids contain starch and other materials which stain heavily with uranyl acetate and lead citrate. When the tracheid starts to form the Si layer, the plastid becomes surrounded by an endoplasmic reticulum. While the fate of these compounds is unknown, it can be envisaged that they are used for generation of energy and/or a source of cell wall materials. 

Figure 8. Changes in the structure of cell organellae during cell wall formation. Upper, middle, and lower photographs are Golgi-body, r-ER, and plastid, respectively. Abbreviations are as follows P, primary wall stage S2B, early part of S2 stage S2L, later part of S2 stage.

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