Plasmodesmata: composition,structure and trafficking

Plasmodesmata are highly specialized gatable trans-wall channels that interconnect contiguous cells and function in direct cytoplasm-to-cytoplasm intercellular transport. Computer-enhanced digital imaging analysis of electron micrographs of plasmodesmata has provided new information on plasmodesmatal fine structure. It is now becoming clear that plasmodesmata are dynamic quasi-organelles whose conductivity can be regulated by environmental and developmental signals. New findings suggest that signalling mechanisms exist which allow the plasmodesmatal pore to dilate to allow macromolecular transport. Plant viruses spread from cell to cell via plasmodesmata. Two distinct movement mechanisms have been elucidated. One movement mechanism involves the movement of the complete virus particle along virus-induced tubular structures within a modified plasmodesma. Apparently two virus-coded movement proteins are involved. A second movement mechanism involves the movement of a non-virion form through existing plasmodesmata. In this mechanism, the viral movement protein causes a rapid dilation of existing plasmodesmata to facilitate protein and nucleic acid movement. Techniques for the isolation of plasmodesmata have been developed and information on plasmodesma-associated proteins is now becoming available. New evidence is reviewed which suggests that plasmodesmatal composition and regulation may differ in different cells and tissues.     

植物转录因子的胞间运动

植物体的组织和器官由多细胞组成,细胞之间的通信对植物体的生长发育必不可少。转录因子作为一类特殊的蛋白质分子不仅在转录水平上参与植物生长发育的调控,而且新近研究发现,转录因子的胞间运动是细胞之间通信方式之一,具有重要的功能。对转录因子胞间运动的发现过程、转录因子胞间运动的机制及其通道进行了论述。转录因子的胞间运动有基于扩散作用的非目标性转运和具有目标性的主动转运两种模式。转录因子胞间运动具有明显的组织特异性和方向性。分析了影响转录因子胞间运动的因素,讨论了转录因子胞间运动的功能以及转录因子胞间运动所参与的植物生长发育及形态建成的调控。     

The distribution of plasmodesmata in the root tip of maize

Summary The distribution of plasmodesmata in different regions of the root apex of Zea mays has been analysed from electron micrographs. There are many more plasmodesmata traversing transverse walls than across longitudinal walls in all the regions studied. When the number of plasmodesmata per unit cell volume is calculated, cells in non-dividing tissue have a considerably lower value than cells in dividing tissue. Evidence for the transport of materials between cells via plasmodesmata is summarised. If it is accepted that plasmodesmata do act as channels for intercellular communication then we believe that their pattern of distribution may be a contributory factor to the process of cell differentiation.     

Ultrastructural analysis of leaf trichome plasmodesmata reveals major differences from mesophyll plasmodesmata

Functional studies on molecular transport through plasmodesmata in leaf mesophyll and trichome cells revealed significant differences in their basal size-exclusion limits and their response to microinjected tobacco mosaic virus movement protein (E. Waigmann et al., 1994, Proc. Natl. Acad. Sci. USA 91: 1433–1437; E. Waigmann and P. Zambryski, 1995, Plant Cell 7: 2069–2079). To address the basis for these functional differences, Nicotiana clevelandii trichome and mesophyll plasmodesmata were compared ultrastructurally. Trichome plasmodesmata increase in ultrastructural complexity from the tip to the base cell. Their neck regions, thought to control molecular traffic through plasmodesmata, are clearly distinct from necks of mesophyll plasmodesmata. In contrast to the electron-dense desmotubular area in mesophyll plasmodesmata, trichome plasmodesmata contain an electron-translucent circle in their center, surrounded by an electron-dense ring. This latter ring is connected to the inner leaflet of the plasma membrane by multiple spokes or filaments. Two monoclonal antibodies raised against a maize plasmodesmal protein preparation (A. Turner et al., 1994, J Cell Sci. 107: 3351–3361) interact with both trichome and mesophyll N. clevelandii plasmodesmata. Based on the localization pattern and the high degree of cross-reactivity, both antibodies likely recognize a conserved structural component of plasmodesmata, and may be useful to mark plasmodesma in a variety of plants and tissues. Received: 24 January 1997 / Accepted: 3 March 1997     

MPB2C,a microtubule-associated plant protein binds to and interferes with cell-to-cell transport of tobacco mosaic virus movement protein

The movement protein of tobacco mosaic virus, MP30, mediates viral cell-to-cell transport via plasmodesmata. The complex MP30 intra- and intercellular distribution pattern includes localization to the endoplasmic reticulum, cytoplasmic bodies, microtubules, and plasmodesmata and likely requires interaction with plant endogenous factors. We have identified and analyzed an MP30-interacting protein, MPB2C, from the host plant Nicotiana tabacum. MPB2C constitutes a previously uncharacterized microtubule-associated protein that binds to and colocalizes with MP30 at microtubules. In vivo studies indicate that MPB2C mediates accumulation of MP30 at microtubules and interferes with MP30 cell-to-cell movement. In contrast, intercellular transport of a functionally enhanced MP30 mutant, which does not accumulate and colocalize with MP30 at microtubules, is not impaired by MPB2C. Together, these data support the concept that MPB2C is not required for MP30 cell-to-cell movement but may act as a negative effector of MP30 cell-to-cell transport activity.     

Subcellular localization determines the availability of non-targeted proteins to plasmodesmatal transport

BACKGROUND: Individual plant cells are encased in a cell wall. To enable cell-to-cell communication, plants have evolved channels, termed plasmodesmata, to span thick walls and interconnect the cytoplasm between adjacent cells. How macromolecules pass through these channels is now beginning to be understood. RESULTS: Using two green fluorescent protein (GFP) reporters and a non-invasive transfection system, we assayed for intercellular macromolecular traffic in leaf epidermal cells. Plasmodesmata were found in different states of dilation. We could distinguish two forms of protein movement across plasmodesmata, non-targeted and targeted. Although leaves have generally been considered closed to non-specific transport of macromolecules, we found that 23% of the cells had plasmodesmatal channels in a dilated state, allowing GFP that was not targeted to plasmodesmata to move into neighboring cells. GFP fusions that were targeted to the cytoskeleton or to the endoplasmic reticulum did not move between cells, whereas those that were localized to the cytoplasm or nucleus diffused to neighboring cells in a size-dependent manner. Superimposed upon this non-specific exchange, proteins that were targeted to the plasmodesmata could transit efficiently between 62% of transfected cells. CONCLUSIONS: A significant population of leaf cells contain plasmodesmata in a dilated state, allowing macromolecular transport between cells. Protein movement potential is regulated by subcellular address and size. These parameters of protein movement illustrate how gradients of signaling macromolecules could be formed and regulated, and suggest that non-cell-autonomous development in plants may be more significant than previously assumed.     

Tobacco mosaic virus movement protein associates with the cytoskeleton in tobacco cells.

Tobacco mosaic virus movement protein P30 complexes with genomic viral RNA for transport through plasmodesmata, the plant intercellular connections. Although most research with P30 focuses on its targeting to and gating of plasmodesmata, the mechanisms of P30 intracellular movement to plasmodesmata have not been defined. To examine P30 intracellular localization, we used tobacco protoplasts, which lack plasmodesmata, for transfection with plasmids carrying P30 coding sequences under a constitutive promoter and for infection with tobacco mosaic virus particles. In both systems, P30 appears as filaments that colocalize primarily with microtubules. To a lesser extent, P30 filaments colocalize with actin filaments, and in vitro experiments suggested that P30 can bind directly to actin and tubulin. This association of P30 with cytoskeletal elements may play a critical role in intracellular transport of the P30-viral RNA complex through the cytoplasm to and possibly through plasmodesmata.     

Getting connected: actin-based cell-to-cell channels in plants and animals

It has been known for more than one hundred years that plant cells are interconnected by cytoplasmic channels called plasmodesmata. This supracellularity was generally considered to be an exotic feature of walled plants containing immobile cells that are firmly enclosed within robust walls. Unexpectedly, intercellular channels in mobile animal cells have been discovered recently. These are extremely dynamic and sensitive to mechanical stress, which causes their rapid breakage and retraction. Both plasmodesmata and nanotubular cell-to-cell channels are supported by the actin cytoskeleton and exclude microtubules. In this article, we discuss the relevance of cell-to-cell channels not only for intercellular communication but also for the development and morphogenesis of multicellular organisms. We also suggest possible parallels between the cell-to-cell transport of endosomes and intracellular pathogens.     

Plasmodesmata: the battleground against intruders

Plasmodesmata are intercellular channels that establish a symplastic communication pathway between neighboring cells in plants. Owing to this role, opportunistic microbial pathogens have evolved to exploit plasmodesmata as gateways to spread infection from cell to cell within the plant. However, although these pathogens have acquired the capacity to breach the plasmodesmal trafficking pathway, plants are unlikely to relinquish control over a structure essential for their survival so easily. In this review, we examine evidence that suggests plasmodesmata play an active role in plant immunity against viral, fungal and bacterial pathogens. We discuss how these pathogens differ in their lifestyles and infection modes, and present the defense strategies that plants have adopted to prevent the intercellular spread of an infection.     

Untersuchungen zur Phytophthora-Prognose

Plasmodesmata are cytoplasmic bridges in plants through which intercellular communication occurs. This involves the transport of ions, photoassimilates, growth hormones as well as protein‐nucleic acid complexes. Although these molecules are rather small (< 1 kDa) plant viruses succeed in using these intercellular highways to transport their genome. These viruses alter the plasmodesmata in some way to allow the transport of such large molecules. This review deals with how plant viruses manage this with the help of movement proteins and the cytoskeleton.     

On the Validity of Plasmodesmograms*

Because the operation of plasmodesmata is of paramount importance for the integrative action of tissues and organs, it is important to quantify the symplastic continuity. An attempt to visualize the intercellular communication pathways in plant tissues is the plasmodesmogram, a two-dimensional diagram of plasmodesmatal frequencies. Plasmodesmograms assume that the greater the frequency of plasmodesmata, the greater is the potential for symplasmic communication. In fact, however, the validity of plasmodesmograms hinges on the premise that plasmodesmata are uniform in structure and functioning. In view of recent findings that plasmodesmata are gatable channels with different functional diameters, a correlation between plasmodesmatal frequency and the rate of intercellular communication - including transport - is disputable and the concept of the plasmodesmogram needs to be re-examined. Its validity appears to vary with the developmental stage and the nature of the plant tissue. Our over-all conclusion is that plasmodesmograms are an acceptable device for coarse (qualitative) assessment of intercellular transport pathways in mature phloem tissues. Pathways identified by plasmodesmograms often correlate with the physiologically determined modes of photosynthate transport. Whether this applies to differentiated tissues, in general, remains to be elucidated. The value of plasmodesmograms for developmental phyisology seems to be limited, as developmental processes coincide with temporary or permanent closure of plasmodesmata.     

Plasmodesmal-associated protein kinase in tobacco and Arabidopsis recognizes a subset of non-cell-autonomous proteins

Cell-to-cell communication in plants involves the trafficking of macromolecules through specialized intercellular organelles, termed plasmodesmata. This exchange of proteins and RNA is likely regulated, and a role for protein phosphorylation has been implicated, but specific components remain to be identified. Here, we describe the molecular characterization of a plasmodesmal-associated protein kinase (PAPK). A 34-kD protein, isolated from a plasmodesmal preparation, exhibits calcium-independent kinase activity and displays substrate specificity in that it recognizes a subset of viral and endogenous non-cell-autonomous proteins. This PAPK specifically phosphorylates the C-terminal residues of tobacco mosaic virus movement protein (TMV MP); this posttranslational modification has been shown to affect MP function. Molecular analysis of purified protein established that tobacco (Nicotiana tabacum) PAPK is a member of the casein kinase I family. Subcellular localization studies identified a possible Arabidopsis thaliana PAPK homolog, PAPK1. TMV MP and PAPK1 are colocalized within cross-walls in a pattern consistent with targeting to plasmodesmata. Moreover, Arabidopsis PAPK1 also phosphorylates TMV MP in vitro at its C terminus. These results strongly suggest that Arabidopsis PAPK1 is a close homolog of tobacco PAPK. Thus, PAPK1 represents a novel plant protein kinase that is targeted to plasmodesmata and may play a regulatory role in macromolecular trafficking between plant cells.     

Targeting and modification of prokaryotic cell–cell junctions by tobacco mosaic virus cell‐to‐cell movement protein

The movement protein (MP) of tobacco mosaic virus (TMV) facilitates the cell-to-cell spread of infection by altering the structure and function of plasmodesmata, the intercellular communication channels in plants. Because the protein was shown to interfere with intercellular communication when expressed in the cyanobacterium Anabaena sp. strain PCC 7120, whether the ability of the protein to target and to modify intercellular communication channels in plants is conserved in this prokaryote was investigated. It was found that the MP localizes to the cell junctions and induces the formation of filamentous structures that traverse the septa. It is proposed that the protein interacts with host components that are similar between plants and Anabaena and that may be evolutionarily related. The observations in Anabaena suggest that the MP modifies plasmodesmata by forming a filamentous aggregate within the pore.     

Transport of macromolecules through plasmodesmata and the phloem

Cell-to-cell communication is a pivotal process in the determination of cell fate during development and physiological adaptation in response to environmental stimuli. The intercellular trafficking of proteins and RNAs has emerged as a novel mechanism of cell-to-cell signaling in plants. As a strategy for efficient intercellular communication, plants have evolved plant-specific symplasmic communication networks via plasmodesmata (PD) and the phloem. PD are symplasmic channels connecting the cytoplasm of neighboring cells and are responsible for the local exchange of metabolites and signaling molecules. The phloem is the sieve-tube system that allows rapid, long-distance translocation of molecules. Together, PD and phloem conduits have been shown to allow the transport of proteins and RNAs in non-selective or/and selective modes. This review describes the current understanding of macromolecule trafficking through PD and the phloem.     

Redox states of plastids and mitochondria differentially regulate intercellular transport via plasmodesmata

Recent studies suggest that intercellular transport via plasmodesmata (PD) is regulated by cellular redox state. Until now, this relationship has been unclear, as increased production of reactive oxygen species (ROS) has been associated with both increased and decreased intercellular transport via PD. Here, we show that silencing two genes that both increase transport via PD, INCREASED SIZE EXCLUSION LIMIT1 (ISE1) and ISE2, alters organelle redox state. Using redox-sensitive green fluorescent proteins targeted to the mitochondria or plastids, we show that, relative to wild-type leaves, plastids are more reduced in both ISE1- and ISE2-silenced leaves, whereas mitochondria are more oxidized in ISE1-silenced leaves. We further show that PD transport is positively regulated by ROS production in mitochondria following treatment with salicylhydroxamic acid but negatively regulated by an oxidative shift in both chloroplasts and mitochondria following treatment with paraquat. Thus, oxidative shifts in the mitochondrial redox state positively regulate intercellular transport in leaves, but oxidative shifts in the plastid redox state counteract this effect and negatively regulate intercellular transport. This proposed model reconciles previous contradictory evidence relating ROS production to PD transport and supports accumulating evidence that mitochondria and plastids are crucial regulators of PD function.     

Non-targeted and targeted protein movement through plasmodesmata in leaves in different developmental and physiological states

Plant cells rely on plasmodesmata for intercellular transport of small signaling molecules as well as larger informational macromolecules such as proteins. A green fluorescent protein (GFP) reporter and low-pressure microprojectile bombardment were used to quantify the degree of symplastic continuity between cells of the leaf at different developmental stages and under different growth conditions. Plasmodesmata were observed to be closed to the transport of GFP or dilated to allow the traffic of GFP. In sink leaves, between 34% and 67% of the cells transport GFP (27 kD), and between 30% and 46% of the cells transport double GFP (54 kD). In leaves in transition transport was reduced; between 21% and 46% and between 2% and 9% of cells transport single and double GFP, respectively. Thus, leaf age dramatically affects the ability of cells to exchange proteins nonselectively. Further, the number of cells allowing GFP or double GFP movement was sensitive to growth conditions because greenhouse-grown plants exhibited higher diffusion rates than culture-grown plants. These studies reveal that leaf cell plasmodesmata are dynamic and do not have a set size exclusion limit. We also examined targeted movement of the movement protein of tobacco mosaic virus fused to GFP, P30::GFP. This 58-kD fusion protein localizes to plasmodesmata, consistently transits from up to 78% of transfected cells, and was not sensitive to developmental age or growth conditions. The relative number of cells containing dilated plasmodesmata varies between different species of tobacco, with Nicotiana clevelandii exhibiting greater diffusion of proteins than Nicotiana tabacum.     

Long distance transport and movement of RNA through the phloem

Cell-to-cell communication is essential for plant development and adaptation to environmental changes. As a strategy for efficient intercellular communication, plants have evolved a plant-specific symplasmic network connected via plasmodesmata that allows a locally restricted information exchange from cell to cell. A rapid information transfer over long distances is enabled via the phloem transport tubes that pervade the complete plant and thus connect even the most distant organs. While communication by small molecules like metabolites and phytohormones is comparably well studied, the intercellular trafficking of proteins and RNAs has only recently emerged as a novel mechanism of cell-to-cell and long-distance signalling in plants. In particular the non-cell-autonomous and systemic transport pathway for specific RNAs seems to play a key role in the co-ordination of important physiological processes, including virus defence, gene silencing, regulation of development, and nutrient allocation. This review is a summary of the current knowledge on RNAs contained in the phloem long-distance transport system, their transport mechanism, and their potential functions.