Arabinogalactan Protein Epitopes Research Articles

Specific Localization of Arabinogalactan-Protein Epitopes at the Surface of Maize Root Hairs

Specific Localization of Arabinogalactan-Protein Epitopes at the Surface of Maize Root Hairs

Cell-specific expression of two arabinogalactan protein epitopes recognized by monoclonal antobodies JIM8 and JIM13 in maize roots

The cell-specific expression of two arabinogalactan protein (AGP) epitopes recognized by monoclonal antibodies JIM8 and JIM13 is reported in maize roots. Employing immunofluorescence and immunogold electron microscopy, the JIM8 antibody was shown to label exclusively protophloem sieve elements, while the JIM13 antibody labelled sieve elements very strongly and adjacent pericycle and companion cells, as well as sloughing root cap cells less strongly. Since the labelling of sieve elements with JIM8 antibody was specific and did not spread to other cell types during root development, it is concluded that this AGP epitope can serve as a specific marker of these specialized cells within the maize root. In the case of the AGP epitope recognized by JIM13 antibody, part of the immunofluorescence label was also found to be associated with cytoplasmic strands in the pericycle and sloughing root cap cells. Immunogold-labelling of sieve elements revealed the association of both AGP epitopes (JIM8 and JIM13) with cortical sieve element reticulum and plasma membranes. Labelling of sieve element reticulum was prominent at its domains of adhesion to the plasma membrane, P-type plastids, and mitochondria. Based on our subcellular studies, we propose a new function of AGP epitopes in endomembrane recognition and adhesion within the sieve elements of maize roots.

Secondary thickening in roots of Arabidopsis thaliana: anatomy and cell surface changes.

The root of Arabidopsis thaliana (L.) Heynh. undergoes dramatic morphological changes during secondary thickening. We have used monoclonal antibodies that recognize two cell surface arabinogalactan protein epitopes (AGPs) and a pectic polysaccharide to document cell surface changes during the process of secondary thickening. These antibodies recognize cells in various stages of differentiation. An AGP epitope recognized by JIM 14 is expressed at the plasma membrane of most tells in the root, but is most highly expressed in mature sieve tube elements, JIM 13 identifies an epitope that is expressed in a discrete ring of cells in the periderm and during xylem vessel element differentiation, disappearing as lignificntion of the vessel elements proceeds, These antibodies reveal a biochemical differentiation at the surface of cells which mirrors obvious morphological differentiation events. Since AGPs have been shown previously to have a dramatic effect on the developmental capabilities of cells it is possible that the epitopes recognized by these antibodies might have an important developmental role during differentiation.

An AGP epitope distinguishes a central metaxylem initial from other vascular initials in theArabidopsis root

The developmental expression of an arabinogalactan protein (AGP) recognised by a monoclonal antibody, JIM 13, is described inArabidopsis roots. It is expressed in the single initial of the central metaxylem vessel that lies immediately above the four central cells of the quiescent centre. AGP expression spreads in a non-clona] fashion to neighbouring cell files as they mature. The AGP first appears in other pre-metaxylem elements and subsequently in the layer of parenchyma cells on either side of the xylem axis. Later, it spreads to the entire ring of eight endodermal cell files and to those pericycle cell files next to the xylem pole. It is postulated that this epitope reflects an underlying set of cell signalling events that may be involved in defining local positional values of cells important in establishing cellular patterns in the root.

Arabinogalactan proteins in maize coleoptiles: developmental relationship to cell death during xylem differentiation but not to extension growth†

The spatial and temporal expression of arabinogalactan proteins (AGPs) in the coleoptile of maize seedlings was investigated with monoclonal antibodies (MAC207, JIM13, JIM14) raised against particular AGP epitopes in carrot. MAC207 binds to a buffer-soluble AGP fraction of 90-210 kDa that also reacts with beta-glucosyl Yariv reagent and the lectin RCA120. Immunogold-labelling showed that the MAC207 epitope is exclusively localized in the plasma membrane. JIM13 binds to a 120 kDa component of the buffer-soluble AGP fraction localized in the plasma membrane of future sclerenchyma cells and secondary-wall thickenings of future tracheids of vascular bundles. JIM14 binds to a 50 kDa component of the salt-extractable fraction from cell walls localized in the innermost wall layer of sclerenchyma cells. These AGP epitopes demonstrate different temporal expression patterns which do not correlate with extension growth. Auxin had no effect on the amount of soluble AGP from coleoptile sections, containing the growth-controlling epidermis but no vascular bundles, as measured by crossed electrophoresis. Moreover, incorporation of radioactive arabinose, galactose or proline into this fraction was not stimulated by auxin. These results contradict the hypothesis that AGPs function as wall-loosening agents in auxin-mediated extension growth. The results are compatible with the notion that AGPs serve as developmental markers defining particular features of future cell differentiation. The specific association of the epitopes recognized by JIM13 and JIM14 with disintegrating cells suggests that the related AGPs identify those cells of the coleoptile which are committed to programmed cell death.

Cell adhesion, cell separation and plant morphogenesis

A simplified and useful view of plant cell development is that it involves the spatial and temporal organization of cell division, cell expansion and cell differentiation all taking place within the constraints of a lack of cell migration. A facet of such development that has often been neglected, is understanding how the cells of a developing structure hold fast or adhere. Subsequently in development the adhered cells of apices or embryos give rise to more loosely attached cells in mature structures where the degree of cell separation and intercellular space formation is developmentally regulated. Recent cell biological studies, most significantly in the derivation of monoclonal antibody probes forthe plant cell surface (Knox, 1992; Roberts, 1990), have increased our awareness of the local modifications of cell walls and rekindled interest in aspects of plant cell wall adhesion and intercellular space formation. It seems timely to examine our current understanding of the mechanisms of cell adhesion and separation and their importance for plant morphogenesis. Molecular probes for cell surface components offer immense potential, not only for the specific study of the local wall modifications such as those resulting in cell separation but also as probes of molecular markers of cell identity relating to position and context. This essay attempts to draw some of the threads together regarding both the nature and importance of cell adhesion and cell separation in plant tissues.

Developmental Regulation of a Plasma Membrane Arabinogalactan Protein Epitope in Oilseed Rape Flowers.

We have identified and characterized the temporal and spatial regulation of a plasma membrane arabinogalactan protein epitope during development of the aerial parts of oilseed rape using the monoclonal antibody JIM8. The JIM8 epitope is expressed by the first cells of the embryo and by certain cells in the sexual organs of flowers. During embryogenesis, the JIM8 epitope ceases to be expressed by the embryo proper but is still found in the suspensor. During differentiation of the stamens and carpels, expression of the JIM8 epitope progresses from one cell type to another, ultimately specifying the endothecium and sperm cells, the nucellar epidermis, synergid cells, and the egg cell. This complex temporal sequence demonstrates rapid turnover of the JIM8 epitope. There is no direct evidence for any cell-inductive process in plant development. However, if cell-cell interactions exist in plants and participate in flower development, the JIM8 epitope may be a marker for one set of them.

Developmental Regulation of a Plasma Membrane Arabinogalactan Protein Epitope in Oilseed Rape Flowers

Developmental Regulation of a Plasma Membrane Arabinogalactan Protein Epitope in Oilseed Rape Flowers

Patterns of expression of the JIM4 arabinogalactan-protein epitope in cell cultures and during somatic embryogenesis in Daucus carota L.

Spatiotemporal patterns of expression of the cell-surface arabinogalactan-protein epitope defined by monoclonal antibody JIM4 (J.P. Knox et al., 1989, Development 106, 47-56) have been characterized by indirect immunofluorescence during the process of somatic embryogenesis in Daucus carota L. The JIM 4 epitope (J4e) occurred on cells established in culture from hypocotyl explants which appeared to derive, at least in part, from the epidermal cells of the hypocotyl. Cultures maintained in the presence of 2,4-dichlorophenoxyacetic acid developed proembryogenic masses of which only infrequent cells at the surface expressed J4e. Sub-culture at a low cell density and withdrawl of the synthetic auxin resulted in an increase in J4e expression in most surface cells and most abundantly in surface layers of cells at the future shoot end of developing embryos. The transition to heart-shaped embryos occurred concurrently with the expression of J4e by groups of cells beneath the developing cotyledons, at the junction of the future root and shoot. At this stage, J4e was also expressed by a single well-defined layer of cells at the surface of the embryos. Advancement to the mature torpedo stage was accompanied by the expression of the epitope on cells forming two regions of the future stele and of cells associated with the cotyledonary provascular tissue characteristic of the carrot seedling. At this stage there was substantially less expression of the marker antigen by epidermal cells, although infrequent expression by isolated cells of the epidermis was maintained. The correlation of J4e expression with the development and distinction of plant tissue patterns during somatic embryogenesis indicates a role for plasma-membrane arabinogalactan proteins in these processes.