N. N. Ibragimova

Specific type of secondary cell wall formed by plant fibers

The review sums data indicating that, in many plant fibers, the secondary cell wall contains so-called gelatinous layers of peculiar structure along with those of common (xylan) structure. Sometimes these gelatinous layers comprise the main bulk of the cell wall. Key characteristics of gelatinous cell wall are presented and compared with those of classic xylan-type cell wall. The process of gelatinous cell wall formation is considered in detail for flax phloem fibers; several characteristic features of this process were revealed: intense rearrangement of already deposited cell-wall layers, unusual dynamics of Golgi vesicles, the occurrence of the stage-specific polysaccharide with specific properties, high activity of β-galactosidase, and the presence of substantial amount of free galactose. Similarity and differences in the gelatinous cell wall formation in the fibers of various plant species are discussed.

Arrangement of mixed-linkage glucan and glucuronoarabinoxylan in the cell walls of growing maize roots.

BACKGROUND AND AIMS Plant cell enlargement is unambiguously coupled to changes in cell wall architecture, and as such various studies have examined the modification of the proportions and structures of glucuronoarabinoxylan and mixed-linkage glucan in the course of cell elongation in grasses. However, there is still no clear understanding of the mutual arrangement of these matrix polymers with cellulose microfibrils and of the modification of this architecture during cell growth. This study aimed to determine the correspondence between the fine structure of grass cell walls and the course of the elongation process in roots of maize (Zea mays). METHODS Enzymatic hydrolysis followed by biochemical analysis of derivatives was coupled with immunohistochemical detection of cell wall epitopes at different stages of cell development in a series of maize root zones. KEY RESULTS Two xylan-directed antibodies (LM11 and ABX) have distinct patterns of primary cell wall labelling in cross-sections of growing maize roots. The LM11 epitopes were masked by mixed-linkage glucan and were revealed only after lichenase treatment. They could be removed from the section by xylanase treatment. Accessibility of ABX epitopes was not affected by the lichenase treatment. Xylanase treatment released only part of the cell wall glucuronoarabinoxylan and produced two types of products: high-substituted (released in polymeric form) and low-substituted (released as low-molecular-mass fragments). The amount of the latter was highly correlated with the amount of mixed-linkage glucan. CONCLUSIONS Three domains of glucuronoarabinoxylan were determined: one separating cellulose microfibrils, one interacting with them and a middle domain between the two, which links them. The middle domain is masked by the mixed-linkage glucan. A model is proposed in which the mixed-linkage glucan serves as a gel-like filler of the space between the separating domain of the glucuronoarabinoxylan and the cellulose microfibrils. Space for glucan is provided along the middle domain, the proportion of which increases during cell elongation.

Free galactose and galactosidase activity in the course of flax fiber development

Composition and content of free monosaccharides and β-galactosidase activity were determined in the course of development of the flax (Linum usitatissimum L.) tissues containing bast fibers. In the stem regions where the fibers were at the stage of formation of the secondary cell wall of gelatinous type, the content of free galactose was high (14 mM) and 13–20 times greater than in the upper part of the stem where the fibers were at the stage of intrusive growth. Pulse-chase experiments demonstrated the differences in the metabolism of individual low-molecular sugars. In respect to glucose and sucrose, all the examined characteristics (content, absolute and specific radioactivity, and the temporal changes of these indices) were identical in the stem regions wherein the fibers were at different stages of development. Labeled galactose was detected only in the stem regions where the fibers were at the stage of secondary cell wall formation. The specific radioactivity of glucose and sucrose reached the maximum immediately after photosynthesis in the presence of 14CO2 and changed in the same way as in the primary products of photosynthesis. The time-course of label incorporation into galactose indicated that this monosaccharide arose as a result of hydrolytic processes. At the stage of secondary cell wall formation, high activity of β-galactosidase was observed, with tissue- and stage-specific fiber β-1,4-galactan as its substrate.

Galactosidase of plant fibers with gelatinous cell wall: Identification and localization

Using a comprehensive approach, we have identified a tissue-specific β-galactosidase from flax (Linum usitatissimum L.) phloem fibers forming a gelatinous cell wall. It was found that when fibers started to develop gelatinous cell wall, β-galactosidase gene expression was enhanced.. Using the antibodies against β-galactosidase, we showed that the enzyme was located in flax phloem fibers where it was detected together with tissue-specific galactan in secreted Golgi vesicles and in gelatinous secondary cell wall. Similar β-galactosidase present in gelatinous cell wall of fibers was found in plants belonging to various taxa and produced by different meristems; these data presume the identical mechanisms of gelatinous cell wall formation and an important role of β-galactosidase. The role of this enzyme in developing the supramolecular structure of gelatinous cell wall is discussed.

Cell wall proteins of flax phloem fibers

A cell wall has been isolated from single-type cells, phloem fibers of flax (Linum usitatissimum L.) being at the stage of the active formation of a thick secondary cell wall. Weakly bound proteins of the cell wall of phloem fibers were extracted and separated, and their mass spectra were recorded. The identification of the proteins and their assignment to a particular cell compartment were performed using a variety of bioinformatics methods. In all, 93 proteins were identified of which many proteins were defined as predicted, putative, and hypothetical. Twenty one proteins were identified as cell-wall proteins. The absence of the marker proteins of primary cell walls such as xyloglucan endotransglycosylase and expansins indirectly confirms the predominance of the secondary cell wall in a sample for protein extraction.

Rhizogenesis in buckwheat thin-cell-layer explants: effect of plant oligosaccharides

Abstract Thin cell-layer explants from Fagopyrum esculentum Moench hypocotyls cultured in liquid medium without growth regulators have been shown to be able to form roots. The intensity of this process depended upon the size of the explant and the length of the hypocotyl. Six diverse oligosaccharide fractions separated from the pea shoot tissues differently effected the rhizogenesis in this system. One fraction from the cell wall pectin and two from the cell sap stimulated this process. Another fraction from the cell wall pectin and two other from the cell sap inhibited explant rooting. One fraction from the cell sap differently influenced two separate stages of root development. These results indicate that the oligosaccharide fragments can be regulators of rhizogenesis.

Cellulosic fibres of flax recruit both primary and secondary cell wall cellulose synthases during deposition of thick tertiary cell walls and in the course of graviresponse

Cellulose synthesising complex consists of cellulose synthase (CESA) subunits encoded by a multigene family; different sets of CESA genes are known to be expressed during primary and secondary cell wall formation. We examined the expression of LusCESAs in flax (Linum usitatissimum L.) cellulosic fibres at various stages of development and in the course of graviresponse by means of RNA-Seq and quantitative PCR. Transcripts for both primary and secondary cell wall-related CESAs were abundant in fibres depositing highly cellulosic tertiary cell walls. Gravistimulation of flax plants temporally increased the abundance of CESA transcripts, specifically in phloem fibres located at the pulling stem side. Construction of coexpression networks for LusCESAs revealed that both primary and secondary cell wall-related CESAs were involved in the joint coexpression group in fibres depositing tertiary cell walls, as distinct from other tissues, where these genes were within separate groups. The obtained data suggest that fibres depositing tertiary cell walls have a specific mechanism of cellulose biosynthesis and a specific way of its regulation.

Development of gravitropic response: unusual behavior of flax phloem G-fibers

The major mechanism of gravitropism that is discussed for herbal plants is based on the nonuniform elongation of cells located on the opposite stem sides, occurring in the growing zone of an organ. However, gravitropic response of flax (Linum usitatissimum L.) is well-pronounced in the lower half of developing stem, which has ceased elongation long in advance of plant inclination. We have analyzed the stem curvature region by various approaches of microscopy and found the undescribed earlier significant modifications in primary phloem fibers that have constitutively developed G-layer. In fibers on the pulling stem side, cell portions were widened with formation of “bottlenecks” between them, leading to the “sausage-like” shape of a cell. Lumen diameter in fiber widening increased, while cell wall thickness decreased. Callose was deposited in proximity to bottlenecks and sometimes totally occluded their lumen. Structure of fiber cell wall changed considerably, with formation of breaks between G- and S-layers. Thick fibrillar structures that were revealed in fiber cell wall by light microscopy got oblique orientation instead of parallel to the fiber axis one in control plants. The described changes occurred at various combinations of gravitational and mechanical stimuli. Thus, phloem fibers with constitutively formed gelatinous cell wall, located in nonelongating parts of herbal plant, are involved in gravitropism and may become an important element in general understanding of the gravity effects on plants. We suggest flax phloem fibers as the model system to study the mechanism of plant position correction, including signal perception and transduction.

Phloem fibres as motors of gravitropic behaviour of flax plants: level of transcriptome

Restoration of stem vertical position after plant inclination is a widely spread version of plant orientation in accordance with gravity vector direction. Gravitropic behaviour of flax plants involves the formation of curvature in stem region that has ceased elongation long in advance of stem inclination. The important participants of such behaviour are phloem fibres with constitutively formed tertiary cell wall (G-layer). We performed the large-scale transcriptome profiling of phloem fibres isolated from pulling and opposite sides of gravitropic curvature and compared with control plant fibres. Significant changes in transcript abundance take place for genes encoding proteins of several ion channels, transcription factors and other regulating elements. The largest number of upregulated genes belonged to the cell wall category; many of those were specifically upregulated in fibres of pulling stem side. The obtained data permit to suggest the mechanism of fibre participation in gravitropic reaction that involves the increase of turgor pressure and the rearrangements of cell wall structure in order to improve contractile properties, and to identify the regulatory elements that operate specifically in the fibres of the pulling stem side making gelatinous phloem fibres an important element of gravitropic response in herbaceous plants.

Screenplay of flax phloem fiber behavior during gravitropic reaction

ABSTRACT Flax phloem fibers act as constitutively formed “muscles” that support the vertical position of the high but narrow stem. The specific mechanical properties of flax fibers and of similar fibers in other plant species are provided by the development of tertiary cell wall with tensed cellulose microfibrils. The work of phloem fibers becomes especially pronounced during the restoration of stem vertical position if it was disturbed. Gravistimulation of flax plants induces considerable modification of phloem fibers at the pulling stem side – the lumen diameter increases, while the cell wall thickness goes down. Here we show that the action of phloem fibers as motors of stem vertical position restoration is coupled to the cell wall remodelling as well as the increase of osmolytes (mainly potassium and malate) content, and accumulation of the γ-amino-butyric acid that may be involved in signalling events. The molecular players that take part in these processes are suggested.