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At least 217 records · Page 12Linked to original sources
PubMed1996

Non-cell-autonomous function of the Antirrhinum floral homeotic proteins DEFICIENS and GLOBOSA is exerted by their polar cell-to-cell trafficking.

In Antirrhinum majus, petal and stamen organ identity is controlled by two MADS-box transcription factors, DEFICIENS and GLOBOSA. Mutations in either of these genes result in the replacement of petals by sepaloid organs and stamens by carpelloid organs. Somatically stable def and glo periclinal chimeras, generated by transposon excision events, were used to study the non-cell-autonomous functions of these two MADS-box proteins. Two morphologically distinct types of chimeras were analysed using genetic, morphological and molecular techniques. Restoration of DEF expression in the L1 cell layer results in the reestablishment of DEF and GLO functions in L1-derived cells only; inner layer cells retain their mutant sepaloid features. Nevertheless, this activity is sufficient to allow the expansion of petal lobes, highlighting the role of DEF in the stimulation of cell proliferation and/or cell shape and elongation when expressed in the L1 layer. Establishment of DEF or GLO expression in L2 and L3 cell layers is accompanied by the recovery of petaloid identity of the epidermal cells but it is insufficient to allow petal lobe expansion. We show by in situ immunolocalisation that the non-cell-autonomy is due to direct trafficking of DEF and GLO proteins from the inner layer to the epidermal cells. At least for DEF, this movement appears to be polar since DEF acts cell-autonomously when expressed in the L1 cell layer. Furthermore, the petaloid revertant sectors observed on second whorl mutant organs and the mutant margins of petals of L2L3 chimeras suggest that DEF and GLO intradermal movement is limited. This restriction may reflect the difference in the regulation of primary plasmodesmata connecting cells from the same layer and secondary plasmodesmata connecting cells from different layers. We propose that control of intradermal trafficking of DEF and GLO could play a role in maintaining of the boundaries of their expression domains.

Biological Transport↗
PubMed1998

Symplasmic fields in the tunica of the shoot apical meristem coordinate morphogenetic events.

In plants, complex cellular interactions, which require the exchange of morphogenetic signals, underlie morphogenesis at the shoot apical meristem. Since all apical meristem cells are interconnected by plasmodesmata, we have investigated if symplasmic paths are available which may preferentially channel metabolites and potential morphogens in the apical meristem, and whether they could support both the formation of determinate appendages and the sustainment of an undifferentiated centre. Experiments in which the permeability of the symplasm was probed with fluorescent dye revealed that the tunica of the apical meristem of birch seedlings (Betula pubescence Ehrh.) is symplasmically compartmentalized into two concentric fields, which restrict the symplasmic diffusion of small potential morphogens to the cells inside their boundaries. A transient connection between the two fields was established early in a plastochron, potentiating the radial exchange of symplasmically diffusing signalling molecules. We suggest that the symplasmic subdivision of the tunica offers a means to unite cells into communication compartments, invoke boundary interactions between them, and shield the distal meristem cells from organogenesis. Electrophysiological measurements indicate that, in addition, the cells of these fields constitute metabolic working units. The relevance of these symplasmic fields for morphogenesis was established experimentally by treatment with short photoperiod, which induced breakdown of the fields into symplasmically isolated cells. Tannic acid staining and in situ immunolocalisation revealed that cell isolation was due to the activation of glucan synthase complexes intrinsic to sphincters. As a result callose plugs were formed on all plasmodesmata leading to morphogenetic deactivation.

Cell Membrane Permeability↗
PubMed1977

Differentiation of the tapetum in Avena. I. The cell surface.

The development of the tapetal cell surface and associated structures in Avena has been followed from cell formation to senescence. Plasmodesmata initially connect the tapetal cells to each other, the pollen mother cells, and the inner loculus wall cells. These connexions are subsequently severed, those to the sporogenous cells being broken first at the pollen mother cell surface during callose wall formation. Loss of cellulose from the tapetal walls was followed using the decline in the ability of the wall to bind the fluorescent brightener, Calcofluor White M2R New. Subplasma-membrane microtubules persist after loss of the cellulose wall. The tapetal plasma membrane facing the meiocytes then develops a series of depressions, or cups, over its surface, which are later the site of pro-orbicule formation. Sporopollenin is laid down over the pro-orbicules, to form orbicules, and over other tapetal cell surfaces. No morphological evidence was found for the intracytoplasmic formation of pro-orbicules or polymerized sporopollenin precursors. These observations on Avena are compared with those on other plants. The changes in the cell wall and associated structures, plasmodesmata and microtubules, are considered in detail, while the general significance of cell wall loss to the water relations of the tissue are assessed. Proposals that pro-orbicule formation results from non-specific accumulation of lipid at a free cell surface are rejected, instead this formation is considered to be related to the presence of a specially modified plasmamembrane surface.

Cell Differentiation↗
PubMed2001

Compartmentation of transport and transfer events in developing seeds.

Developing seeds are net importers of organic and inorganic nutrients. Nutrients enter seeds through the maternal vascular system at relatively high concentrations in the phloem. They exit importing sieve elements via interconnecting plasmodesmata and, during subsequent symplasmic passage, are sequestered into labile storage pools (vacuoles; starch). Transporters function to retrieve nutrients leaked to the seed apoplasm during symplasmic passage. Maternal cells responsible for nutrient release to the seed apoplasm are characteristically located at the maternal/filial interface. Their plasma membranes are enriched in transport proteins and, in some species, these cells are modified to a transfer cell morphology. Apoplasmic volumes of seeds are relatively small, but contain high concentrations of sugars, potassium and a range of amino acids. Sucrose and amino acids are taken up from the seed apoplasm by one to two cell layers of filial tissues that juxtapose the maternal tissues. The plasma membranes of the uptake cells are enriched in sucrose and amino acid/H(+) transporters which co-localize with H(+)-ATPASES: In some species, these cells are modified to a transfer cell morphology. High densities of plasmodesmata support symplasmic delivery of accumulated nutrients to underlying storage cells where polymer formation (starch, protein) takes place. Hexoses, resulting from sucrose hydrolysis and leakage to the seed apoplasm, are retrieved by hexose/H(+) symporters.

Amino Acids↗
PubMed2002

Sensitivity of cell hydraulic conductivity to mercury is coincident with symplasmic isolation and expression of plasmalemma aquaporin genes in growing maize roots.

Root elongation occurs as individual cells along the growing zone increase in volume. This increase is caused by water entering the cell either by moving across the cell membrane from the apoplast via aquaporins, or entering through plasmodesmata that symplastically connect cells to each other or with the sieve element. In this investigation we used mercury, a known inhibitor of aquaporin water channels, to manipulate the water permeability of growing maize root cells. 20 micro M HgCl(2) was found to reduce root elongation by around 75% and this reduction in growth was greatest in the older growing cells, with little effect on the younger cells near the root tip. Cell hydraulic conductivity (Lp) of cells close to the root tip (at 3 mm) remained unaffected by mercury treatment in contrast to older growing and non-growing cells where Lp was greatly reduced. Using reverse transcription-polymerase chain reaction analysis, younger root regions were shown to express higher levels of two plasmalemma intrinsic protein genes than older root regions further away from the root tip. However, a gene encoding a tonoplast aquaporin was expressed at similar levels in both regions of the growing zone. The fluorescent tracer, carboxyfluorescein, demonstrated symplastic connection between the phloem and root cortical cells at 3 mm but not at 5 or 20 mm. The data are consistent with a decrease in symplastic continuity along the growing zone and highlight a change in the principal pathway of water uptake during the development of the growing root cell.

Algorithms↗
PubMed2005

Intrusive growth of flax phloem fibers is of intercalary type.

Flax (Linum usitatissimum L.) phloem fibers elongate considerably during their development and intrude between existing cells. We questioned whether fiber elongation is caused by cell tip growth or intercalary growth. Cells with tip growth are characterized by having two specific zones of cytoplasm in the cell tip, one with vesicles and no large organelles at the very tip and one with various organelles amongst others longitudinally arranged cortical microtubules in the subapex. Such zones were not observed in elongating flax fibers. Instead, organelles moved into the very tip region, and cortical microtubules showed transversal and helical configurations as known for cells growing in intercalary way. In addition, pulse-chase experiments with Calcofluor White resulted in a spotted fluorescence in the cell wall all over the length of the fiber. Therefore, it is concluded that fiber elongation is not achieved by tip growth but by intercalary growth. The intrusively growing fiber is a coenocytic cell that has no plasmodesmata, making the fibers a symplastically isolated domain within the stem.

Cell Enlargement↗
PubMed2003

Symplasmic protein and RNA traffic: regulatory points and regulatory factors.

Plasmodesmata and the phloem form a cytoplasmic network that permits direct cell-cell communication in plants. This network can mediate the trafficking of selective proteins and RNAs that may have important developmental functions. Recent work has provided evidence that protein and RNA traffic across specific interfaces of this network is regulated in a distinct manner. Progress has been made in identifying potential cellular factors that confer such regulation. These advances should promote further investigations into the mechanisms and functions of protein and RNA traffic using biochemical, cellular, genetic and molecular tools.

Cytoplasm↗
PubMed2004

The plasmodesmatal transport pathway for homeotic proteins, silencing signals and viruses.

Non-cell-autonomous signals in the form of microRNAs and transcription factors could have important developmental functions. Plasmodesmata (PD) form a cytoplasmic network throughout the plant body and provide the means of symplasmic cell-to-cell transport in plants. Homeodomain transcription factors, small RNA molecules and viral genomic information move selectively to adjacent cells via PD microchannels. Tissue-specific expression studies of non-cell-autonomous transcription factors and RNA molecules have confirmed that their intercellular transport is a highly regulated process, which depends on the tissue, developmental stage and nature of the transported macromolecule. We have known for some time that gene-silencing signals spread both locally from cell to cell and across long distances following the source to sink transition. Recent work has provided evidence that small single-stranded silencing-induced RNAs and microRNA molecules are present in the phloem transport system of different plant species. Further, recent evidence has confirmed that the transport of silencing RNA via PD is a regulated and active process, and that an amplification-relay mechanism is in place for the long-distance spread of silencing signals.

Animals↗
PubMed2005

Regulation of short-distance transport of RNA and protein.

The intercellular trafficking of proteins and RNAs has emerged as a novel mechanism of cell-cell communication in plant development. Plasmodesmata (PD), intercellular cytoplasmic channels, have a central role in cell-cell trafficking of regulatory proteins and RNAs. Recent studies have demonstrated that plants use either a selective or a non-selective PD trafficking pathway for regulatory proteins. Moreover, plants have developed strategies to regulate both selective and non-selective movement. Recent work has focused especially on integrating the recent understanding of the function and mechanisms of intercellular macromolecule movement through PD.

DNA, Plant↗
PubMed2006

Plant viral movement proteins: agents for cell-to-cell trafficking of viral genomes.

Plants viruses spread throughout their hosts using a number of pathways, the most common being movement cell to cell through plasmodesmata (PD), unique intercellular organelles of the plant kingdom, and between organs by means of the vascular system. Pioneering studies on plant viruses revealed that PD allow the cell-to-cell trafficking of virally encoded proteins, termed the movement proteins (MPs). This non-cell-autonomous protein (NCAP) pathway is similarly employed by the host to traffic macromolecules. Viral MPs bind RNA/DNA in a sequence nonspecific manner to form nucleoprotein complexes (NPC). Host proteins are then involved in the delivery of MPs and NPC to the PD orifice, and a role for the cytoskeleton has been implicated. Trafficking of NCAPs through the PD structure involves three steps in which the MP: (a) interacts with a putative PD docking complex, (b) induces dilation in the PD microchannels, and (c) binds to an internal translocation system for delivery into the neighboring cytoplasm. Viral genera that use this NCAP pathway have evolved a combination of a MP and ancillary proteins that work in concert to enable the formation of a stable NPC that can compete with endogenous NCAPs for the PD trafficking machinery. Incompatible MP-host protein interactions may underlie observed tissue tropisms and restricted infection domains. These pivotal discoveries are discussed in terms of the need to develop a more comprehensive understanding of the (a) three-dimensional structure of MPs, (b) PD supramolecular complex, and (c) host proteins involved in this cell-to-cell trafficking process.

Cell Nucleus↗
PubMed2003

Dispatch. Intercellular signaling: an elusive player steps forth.

Plasmodesmata play a central role in cell-to-cell communication in plants, allowing transport of regulatory proteins and mRNAs. A recent study has identified a specific protein that regulates the intercellular transport of macromolecules in plants, known as non-cell autonomous pathway protein 1.

Gene Expression Regulation↗
PubMed2003

A polarized cell: the root statocyte.

In the gravity-perceiving cells (statocytes), located in the centre of the root cap, polarity is expressed in the arrangement of the organelles since, in most genera, the nucleus and the endoplasmic reticulum are maintained at the opposite ends of each cell by actin. Polarity is also evident in the distribution of plasmodesmata, which are more numerous in the transverse walls than in the longitudinal walls. The centre of each statocyte is depleted of microtubules (they are only located at the periphery) but is occupied by numerous amyloplasts (statoliths), denser than the cytoplasm. The amyloplasts do not contribute to the inherent structural polarity since their position is dependent upon the gravity vector. This article focuses on new microscopic analyses and on data obtained from experiments performed in microgravity, which have contributed to our better understanding of the architecture of the actin web implicated in the perception of gravity. Depending upon the plant, the actin network seems to be formed of single filaments arranged in various ways, or, of thin bundles of actin filaments. The amyloplasts are enmeshed in this web of actin and their envelopes are associated with it, but they can have autonomous movement via myosin in the absence of gravity. From calculations of the value of the force necessary to move one amyloplast in the lentil root, and from videomicroscopy performed with living statocytes of maize roots, it is hypothesized that actin microfilaments could be orientated in an overall diagonal direction in the statocyte. These observations could help in understanding how slight amyloplast movements may trigger and transmit the gravitropic signal.

Actin Cytoskeleton↗
PubMed2005

Cell-to-cell movement of GFP during embryogenesis and early seedling development in Arabidopsis.

There is increasing evidence for intercellular trafficking of macromolecules through plasmodesmata (PD) during plant development. Here we study the ability of PD to traffic proteins during embryogenesis and early seedling development in Arabidopsis. Transgenic lines that induce GFP expression only in meristems, MSG (meristem-specific GFP), were used to monitor GFP movement. Cell-to-cell movement of different-sized GFP reporters reveals that embryos and young seedlings traffic proteins at least 54 kDa in size. Although 27-kDa soluble GFP (1 x sGFP) freely moves between cells throughout the entire embryo during all stages analyzed, 2 x sGFP movement becomes more restricted as development proceeds. After germination, cells near the apical meristem in seedlings show a higher size exclusion limit (SEL), whereas the SEL becomes more restricted as surrounding tissues develop identities. Although 1 x sGFP moves throughout leaf primordia, as the leaf develops only the basal part of leaf petioles, main vascular tissues, and leaf veins (not blades) allow 1 x sGFP movement. Although previous studies showed that embryos allow movement of small symplastic tracers (0.5 kDa), the present data demonstrate that the embryo constitutes a single symplast that allows transport of macromolecules as well. Even 2 x sGFP moves from its site of expression at the apical meristem in embryos and seedlings, yet the extent of movement is more limited than 1 x sGFP. Thus, PD have distinct SELs in different subregions of the embryo and seedling. These studies support the general concept that PD in younger tissues are more dilated and less restrictive than PD in older (nonvascular) tissues.

Arabidopsis↗
PubMed2005

Not just another hole in the wall: understanding intercellular protein trafficking.

Development and differentiation of multicellular organisms requires cell-to-cell communication. In plants direct signaling and exchange of macromolecules between cells is possible through plasmodesmata. Recently direct exchange of membrane-bound vesicles and organelles has been demonstrated between animal cells through formation of cytoplasmic bridges (tunneling nanotubes) in vitro. Here we review recent developments in cell-to-cell trafficking of macromolecules in plants and animals.

Animals↗
PubMed2006

Quantification of plasmodesmatal endoplasmic reticulum coupling between sieve elements and companion cells using fluorescence redistribution after photobleaching.

Transgenic tobacco (Nicotiana tabacum) was studied to localize the activity of phloem loading during development and to establish whether the endoplasmic reticulum (ER) of the companion cell (CC) and the sieve element (SE) reticulum is continuous by using a SUC2 promoter-green fluorescent protein (GFP) construct targeted to the CC-ER. Expression of GFP marked the collection phloem in source leaves and cotyledons as expected, but also the transport phloem in stems, petioles, midveins of sink leaves, nonphotosynthetic flower parts, roots, and newly germinated seedlings, suggesting that sucrose retrieval along the pathway is an integral component of phloem function. GFP fluorescence was limited to CCs where it was visualized as a well-developed ER network in close proximity to the plasma membrane. ER coupling between CC and SEs was tested in wild-type tobacco using an ER-specific fluorochrome and fluorescence redistribution after photobleaching (FRAP), and showed that the ER is continuous via pore-plasmodesma units. ER coupling between CC and SE was quantified by determining the mobile fraction and half-life of fluorescence redistribution and compared with that of other cell types. In all tissues, fluorescence recovered slowly when it was rate limited by plasmodesmata, contrasting with fast intracellular FRAP. FRAP was unaffected by treatment with cytochalasin D. The highest degree of ER coupling was measured between CC and SE. Intimate ER coupling is consistent with a possible role for ER in membrane protein and signal exchange between CC and SE. However, a complete lack of GFP transfer between CC and SE indicated that the intraluminal pore-plasmodesma contact has a size exclusion limit below 27 kD.

Cell Communication↗
PubMed2002

Plasmodesma-mediated selective protein traffic between "symplasmically isolated" cells probed by a viral movement protein.

Intercellular communication is essential for differentiation and development. In plants, plasmodesmata (PD) form cytoplasmic channels for direct communication. During plant development, programmed reduction in PD number and transport capacity creates the so-called symplasmic domains. Small fluorescent dyes and ions can diffuse among cells within a domain but not across domain boundaries. Such symplasmic isolation is thought to allow groups of cells to differentiate and develop into tissues with distinct structures and functions. Whether or how "symplasmically isolated" cells communicate with one another is poorly understood. One well-documented symplasmic domain is the sieve element-companion cell (SE-CC) complex in the phloem tissue. We report here that, when produced in the CC of transgenic tobacco, the 3a movement protein (3a MP) of Cucumber mosaic virus fused to green fluorescent protein (GFP) can traffic out of the SE-CC complex via PD. The extent of 3a MP:GFP traffic across the boundary between vascular and nonvascular tissues depends on organ type and developmental stage. Our findings provide experimental evidence that endogenous machinery exists for protein traffic between the symplasmically isolated SE-CC complex and neighboring cells. We suggest that PD-mediated traffic of selected macromolecules can be a mechanism for symplasmically isolated cells to communicate with one another.

Cell Membrane↗
PubMed2004

Direct role of a viroid RNA motif in mediating directional RNA trafficking across a specific cellular boundary.

The plasmodesmata and phloem form a symplasmic network that mediates direct cell-cell communication and transport throughout a plant. Selected endogenous RNAs, viral RNAs, and viroids traffic between specific cells or organs via this network. Whether an RNA itself has structural motifs to potentiate trafficking is not well understood. We have used mutational analysis to identify a motif that the noncoding Potato spindle tuber viroid RNA evolved to potentiate its efficient trafficking from the bundle sheath into mesophyll that is vital to establishing systemic infection in tobacco (Nicotiana tabacum). Surprisingly, this motif is not necessary for trafficking in the reverse direction (i.e., from the mesophyll to bundle sheath). It is not required for trafficking between other cell types either. We also found that the requirement for this motif to mediate bundle sheath-to-mesophyll trafficking is dependent on leaf developmental stages. Our results provide genetic evidence that (1) RNA structural motifs can play a direct role in mediating trafficking across a cellular boundary in a defined direction, (2) the bundle sheath-mesophyll boundary serves as a novel regulatory point for RNA trafficking between the phloem and nonvascular tissues, and (3) the symplasmic network remodels its capacity to traffic RNAs during plant development. These findings may help further studies to elucidate the interactions between RNA motifs and cellular factors that potentiate directional trafficking across specific cellular boundaries.

Base Sequence↗
PubMed2003

Fusion to GFP blocks intercellular trafficking of the sucrose transporter SUT1 leading to accumulation in companion cells.

BACKGROUND: Plant phloem consists of an interdependent cell pair, the sieve element/companion cell complex. Sucrose transporters are localized to enucleate sieve elements (SE), despite being transcribed in companion cells (CC). Due to the high turnover of SUT1, sucrose transporter mRNA or protein must traffic from CC to SE via the plasmodesmata. Localization of SUT mRNA at plasmodesmatal orifices connecting CC and SE suggests RNA transport, potentially mediated by RNA binding proteins. In many organisms, polar RNA transport is mediated through RNA binding proteins interacting with the 3'-UTR and controlling localized protein synthesis. To study mechanisms for trafficking of SUT1, GFP-fusions with and without 3'-UTR were expressed in transgenic plants. RESULTS: In contrast to plants expressing GFP from the strong SUC2 promoter, in RolC-controlled expression GFP is retained in companion cells. The 3'-UTR of SUT1 affected intracellular distribution of GFP but was insufficient for trafficking of SUT1, GFP or their fusions to SEs. Fusion of GFP to SUT1 did however lead to accumulation of SUT1-GFP in the CC, indicating that trafficking was blocked while translational inhibition of SUT1 mRNA was released in CCs. CONCLUSION: A fusion with GFP prevents targeting of the sucrose transporter SUT1 to the SE while leading to accumulation in the CC. The 3'-UTR of SUT1 is insufficient for mobilization of either the fusion or GFP alone. It is conceivable that SUT1-GFP protein transport through PD to SE was blocked due to the presence of GFP, resulting in retention in CC particles. Alternatively, SUT1 mRNA transport through the PD could have been blocked due to insertion of GFP between the SUT1 coding sequence and 3'-UTR.

3' Untranslated Regions↗