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Expression of the crucifer-infecting TMV-Cg movement protein in tobacco plants complements in trans a TMV-U1 trafficking-deficient mutant.

Tobamovirus movement proteins play a determinant role in the establishment of infections in plants, allowing the local movement of viral RNA genome through plasmodesmatas. We expressed the movement protein (MP) of the crucifer- and garlic-infecting Tobacco Mosaic Virus strain Cg (TMV-Cg) in both resistant Xanthi NN and sensitive Xanthi nn Nicotiana tabacum plants. MP-Cg function was assayed by inoculating transgenic plants with a trafficking-deficient mutant of TMV strain U1. Following infection, local necrotic lesions were developed in resistant transgenic plants, and a systemic infection was produced in sensitive tobaccos. Thus, movement function of the mutant virus was complemented in trans by MP-Cg expressed in transgenic plants, causing the same symptoms as wild-type strain. We demonstrated that the function of MP-U1 could be replaced efficiently by MP-Cg, even though these proteins share only 36% of identity. Similar hydrophobic patterns of MP-Cg and MP-U1 suggests structure and function conservations of both proteins. This work is an example of how two tobamoviruses differing in their host range help to understand viral movement mechanism during the infection.

Gene Expression↗

Oxygen transfer characteristics in the compact callus aggregates of Rhodiola sachalinensis.

The effective diffusivity of oxygen in compact callus aggregates (CCA) of Rhodiola sachalinensis was extremely small, varying from 0.34 x 10(-11) to 5.4 x 10(-9) m2/s and increasing with particle diameter. The calculated results indicated that severe oxygen limitations occurred in the CCA aggregates. However, the direct determination of the viability of the cells demonstrated that the CCA aggregates contained a high fraction of viable cells. It was suggested that the tissue differentiation or plasmodesmata within the CCA aggregates may provide capillaries for improving transport of oxygen and other nutrients.

Drugs, Chinese Herbal↗

Further studies of the glandular tissue of the Sauromatum guttatum (Araceae) appendix.

Electron microscopic studies showed that the trans-Golgi network (trans indicates the polarity of cisternae within the Golgi apparatus; it is opposite to the cis-face that is adjacent to the rough endoplasmic reticulum) was involved in the processing of the osmiophilic material present in the appendix of the inflorescence of Sauromatum guttatum. This material accumulated in the rough endoplasmic reticulum and in special pockets of the plasma membrane prior to heat production. Associations between the endoplasmic reticulum and trans-Golgi network were observed. The Golgi apparatus was composed of 5-6 dictyosomes on one side and one or two somewhat detached cisternae on the other side. Various nonosmiophilic Golgi-derived vesicles were observed: small ones covered with spike-like material, large ones with a smooth surface, and irregularly shaped ones. These electron-translucent vesicles seemed to accumulate in specific localities at the plasma membrane surface in the vicinity of the osmiophilic material; they were not found when the aroma was released. During heat production, the Golgi structures shrank and the activity of the trans-Golgi network seemed to be reduced. At the same time, coated pits were seen at the plasma membrane surface. In some cells, hypertrophic Golgi apparatuses were seen with only 2-3 dictyosomes that contained granulated material in their lumens. Finally, the osmiophilic material was also found in the plasmodesmata.

Journal Article↗

From gravity and the organism to gravity and the cell.

This workshop on Gravity and the Cell was modeled on a 1968 conference on Gravity and the Organism. Each conference tried to identify the most salient scientific questions about how gravity is important to living systems. In the roughly two decades between the conferences there have been impressive advances in experimental methods, but the major scientific questions that have driven their applications to problems of gravitational biology, i.e., our broad research goals, remain much the same. In the case of plant research, improvements and extensions of biochemical techniques for investigating the ways organisms use environmental (g-force) information in salubrious ways has kept pace with progress in plant biochemistry. We now know much more about the roles of organic and inorganic substances that plants employ for information transfer and growth regulation and about the avenues and mechanisms for transport of those key substances within organisms. We have seen the acceptance of a "local control" concept that regulation of g-responses depends critically on plasmodesmata and gap junctions which allow plants and animals to throttle the transport of growth regulators across tissue boundaries often, especially in the case of plants, far removed from the morphological regions of concentration of statocytes that were once thought to be the exclusive bioaccelerometers used by plants. NASA's sponsorship of ground-based research in gravitational biology has served as important underpinning for orbital flight programs in space biology. The currently most noteworthy research area of ignorance is the mechanism by which the physical event of g-force susception becomes the biological process of g-force perception. Only rarely has it been possible to perform a definitive test of a theory of mechanism of gravity perception. Therefore, experimental research efforts in gravitational physiology still are essentially dependent on exploratory studies for which stimulus/response experiments require experimental manipulation of test subjects' g-force environment. Many research efforts have focused narrowly on contrasting the responses of test subjects to only two gravitational environments, 1 g versus real or simulated zero g. Much less effort has been devoted to studies at other g levels. For experiments in gravitational physiology three rotating machines have made, and no doubt will continue to make, important contributions: the centrifuge, the clinostat, and, most of all, the orbiting space vehicle. Some research trends, viewpoints, and shifts of emphasis are discussed.

Animals↗

Geoperception in primary and lateral roots of Phaseolus vulgaris (Fabaceae). I. Structure of columella cells.

Primary roots of Phaseolus vulgaris (Fabaceae) are positively geotropic, while lateral roots are not responsive to gravity In order to elucidate the structural basis for this differential georesponse, we have performed a qualitative and quantitative analysis of the ultrastructure of columella cells of primary and lateral roots of P. vulgaris. Root systems were fixed in situ so as not to disturb the ultrastructure of the columella cells. The columellas of primary roots are more extensive than those of lateral roots. The volumes of columella cells of primary roots are approximately twice those of columella cells of lateral roots. However, columella cells of primary roots contain greater absolute volumes and numbers of all cellular components examined than do columella cells of lateral roots. Also, the relative volumes of cellular components in columella cells of primary and lateral roots are statistically indistinguishable. The endoplasmic reticulum is sparse and distributed randomly in both types of columella cells. Both types of columella cells contain numerous sedimented amyloplasts, none of which contact the cell wall or form complexes with other cellular organelles. Therefore, positive geotropism by roots must be due to a factor(s) other than the presence of sedimented amyloplasts alone. Furthermore, it is unlikely that amyloplasts and plasmodesmata form a multi-valve system that controls the movement of growth regulating substances through the root cap.

Endoplasmic Reticulum↗

Rootcap structure in wild type and in a starchless mutant of Arabidopsis.

Rootcaps of the wild type (WT) and of a starchless, gravitropic mutant (TC7) of Arabidopsis thaliana L. were examined by electron microscopy to identify cellular polarities with respect to gravity. In columella cells, nuclei are located proximally, and the nuclear envelope is continuous with endoplasmic reticulum (ER) that is in turn connected to nearby plasmodesmata. Impregnation of ER with osmium ferricyanide revealed numerous contacts between columella plastids and ER in both genotypes. ER is present mostly in the outer regions of the columella protoplast except in older columella cells that are developing into peripheral cells. In vertical roots, only columella cells that are intermediate in development (story 2 cells) have a higher surface density (S) of ER in the distal compared to proximal regions of the cell. The distal but not the proximal S of the ER is constant throughout columella development. Plastids are less sedimented in TC7 columella cells compared to those of the WT. It is hypothesized that plastid contact with the ER plays a role in gravity perception in both genotypes.

Arabidopsis↗

The tomato mosaic tobamovirus movement protein interacts with a putative transcriptional coactivator KELP.

Viral movement through plasmodesmata in host plants likely depends on the interaction between virus-encoded movement protein (MP) and host proteins. In order to search for MP-interacting protein (MIP), we carried out far-western screening of a Brassica campestris cDNA library using a recombinant MP of tomato mosaic tobamovirus (ToMV) as a probe. One of the positive clones, designated MIP102, was found to be a putative orthologue for a transcriptional coactivator KELP of Arabidopsis thaliana. In vitro analysis with recombinant proteins revealed that ToMV MP could bind to KELP proteins that are derived from different plant species. At least 31 amino acids from the carboxyl-terminus of ToMV MP were dispensable for the interaction with KELP. Other MPs, derived from crucifer tobamovirus CTMV-W and cucumber mosaic cucumovirus, also exhibited comparable binding abilities. This suggests that these MPs could commonly interact with KELP, possibly to modulate the host gene expression.

Amino Acid Sequence↗

Cloning of a tobacco cDNA coding for a putative transcriptional coactivator MBF1 that interacts with the tomato mosaic virus movement protein.

Viral movement through plasmodesmata in host plants depends on the interaction between virus-encoded movement protein (MP) and host proteins. To search for MP-interacting protein (MIP), far-western screening of a tobacco cDNA library was carried out using a recombinant MP of tomato mosaic virus (ToMV) as a probe. One of the positive cDNA clones, designated MIP204, was highly homologous to a class of transcriptional coactivators commonly referred to as multiprotein bridging factor 1 (MBF1). ToMV MP could also bind to the Arabidopsis homologues of MBF1. The recombinant MIP204 bound to MPs of ToMV and a crucifer tobamovirus CTMV-W, but not of cucumber mosaic virus. MPs of ToMV and the related virus may interact with MBF1-like proteins to modulate host gene expression.

Arabidopsis Proteins↗

Further ultrastructural research of Chara vulgaris spermiogenesis: endoplasmic reticulum, structure of chromatin, 3H-lysine and 3H-arginine incorporation.

On the basis of morphological features, 10 consecutive structural phases of spermatids were identified in Chara vulgaris spermiogenesis. They were schematically presented. In early and middle spermiogenesis, i.e. during the period preceding formation of fibrillar structure of mature spermatozoid nucleus, a slight remodelling of chromatin, accompanied by proplastid transformation into an amyloplast as well as by development of 2 flagella and a microtubular manchette, is observed. First, condensed chromatin concentrates around the nuclear envelope (phases III-V) and then it transforms into a network-like structure (phase VI). This change in chromatin structure is preceded by nucleolar extrusion to the cytoplasm where nucleoli become degraded (phase IV) and by a dynamic development of rough endoplasmic reticulum (RER) (phase V) which is continuous with the nuclear envelope and with RER of the adjacent spermatids via plasmodesmata. The inner membrane of the nuclear envelope invaginates into the nucleoplasm in which "nuclear reticulum" appears. It all happens during increased 3H-arginine and 3H-lysine incorporation into proteins which are rapidly translocated into the nucleus. In medium-late spermiogenesis (phases VI-VIII), network-like condensed chromatin disappears. Next, the structure of the nucleus changes dramatically. Short, randomly positioned fibrils (phase VII) appear and gradually become longer (phase VIII), thicker (phase IX) and more distinct, lying parallel to the surface of elongating and curling nucleus. Membranes of the nuclear envelope become closer to each other and a distinct dark layer--probably lamin--appears adhering to the inner membrane of the nuclear envelope. Towards the end of spermiogenesis (phase X), very densely packed parallel helices, ca 2 nm in diameter, are visible. The surfaces of flagella and the spermatozoid are covered with diamond-shaped larger and smaller scales, respectively. Helically coiled spermatozoids are liberated from antheridial filament cells through earlier created (phase VIII) "liberation pores" with pads of unknown nature.

Arginine↗

Interaction of tomato mosaic virus movement protein with tobacco RIO kinase.

Tomato mosaic virus (ToMV) has a regulatory gene encoding a movement protein (MP) that is involved in the cell-to-cell movement of viral RNA through plasmodesmata. To identify the host cell factors interacting with ToMV MP, we used a recombinant MP probe to isolate cDNA clones from a phage expression library of Nicotiana tabacum by a far-Western screening method. One of the cDNA clones encoded an MP-interacting protein, MIP-T7, homologous to the yeast novel protein kinase, Rio1p. We isolated a full-length cDNA by RT-PCR. The putative gene product was designated NtRIO, and shared 33 and 73% amino acid identity with yeast and Arabidopsis RIO kinases, respectively. In vitro analyses using recombinant proteins showed that NtRIO also interacted with a different MP derived from Cucumber mosaic virus. NtRIO had autophosphorylation activity and phosphorylated ToMV MP. Addition of recombinant tobacco casein kinase 2 resulted in a marked increase in the phosphorylation of NtRIO. The interaction between NtRIO and ToMV MP was inhibited by phosphorylation of NtRIO.

Amino Acid Sequence↗

Uptake and transport of calcium in plants.

Recently, research on Ca(2+) transport in plants has been focused on cellular and molecular level. But the uptake, transport and distribution are also very important for calcium to accomplish its function at whole plant level. There are many cells along the way of transport of Ca(2+) from root to shoot, and Ca(2+) passes either through the cytoplasm of cells linked by plasmodesmata (the symplast) or through the spaces between cells (the apoplast), which include Ca(2+) uptake by root cells, Ca(2+) transport from root cortex to and through the xylem, and then out of it into leaves or fruits. Ca(2+) channels, Ca(2+)/H(+) antiporter and Ca(2+)-ATPase play roles in the uptake and transport of Ca(2+) in root cells. To be transported from root surface to xylem, Ca(2+) needs to traverse endodermal cells and xylem parenchyma cells. Endodermal Casparian band, the main barrier for the apoplastic movement of ions into the stele, compels some Ca(2+) to enter root symplast through Ca(2+) channels in endodermal cells and then reach xylem parenchyma. Ca(2+)-ATPase may drive Ca(2+) into the stelar apoplast from xylem parenchyma. Some Ca(2+) effuses from endodermal cell and then get to xylem through apoplastic pathway. Ca(2+) is transported in plant xylem vessel in chelate form and the speed of water flow is the key factor Ca(2+) transport via xylem in trunk. There are both apoplastic and symplastic pathways of Ca(2+) transport in fruit or leaf tissue too.

Biological Transport↗

[Ultracytochemical studies of aloin in Aloe arborescens leaves].

Lead acetate precipitation method was used for ultracytochemical localization of aloin. The processes of aloin production, transport and storage were studied by transmission electron microscope. Results showed that aloin was produced in the plastids of the assimilating tissue. The aloin was transported through the plastid membrane to the surrounding endoplasmic reticulum and enveloped in the vesicles by the endoplasmic reticulum elements, the vesicles approached and later fused with the plasmalemma. Some vesicles of the plastid membrane directly fused with the plasmalemma. The vesicles released their contents into the apoplast through exocytosis, and finally reached the vascular bundle sheath by apoplastic translocation. Aloin was transported to the internal tangential wall of vascular bundle sheath cell through endoplasmic reticulum vesicles, and reached the cytoplasm of aloin cell by means of plasmodesmata. Finally, aloin was stored in the vacuoles of aloin cell.

Aloe↗

The cell walls of Chara aspera Willd. (Charophyta) vegetative cells.

The ultrastructure of the vegetative cell walls of the charophyte Chara aspera Willd was studied with TEM. Thallus cells, rhizoid bulbil and rhizoidal node cells were investigated. The internodal cells transverse walls contained plasmodesmata. The longitudinal walls of the internodal cells were uniform, fibrillar, with two thin structurally distinct layers with different structure facing the cytoplasm. The outermost layers of internodal, cortical and rhizoid bulbil cells were composed of randomly orientated fibrils. The longitudinal walls of the cortical cells were helicoidal in structure. In the rhizoid bulbil cell walls, six different layers could be distinguished, but their occurrence seemed to depend on the fixation, staining and cutting procedures. A middle lamella and osmophilic deposits were found in the wall between rhizoidal node cells. The cytoplasmic structure of the internodal and cortical cells was not found to differ from other species of Chara. Charasomes were observed only in cortical cells.

Cell Wall↗

Cellular interactions in tissue and organ development.

Developmental patterns result from combinations of interactions and intracellular programmes. The purpose here is to define the roles of interactions wherever possible and to consider their major parameters, such as the timing of their action, their specificity and the distances over which they occur. The approach is one of a board survey, attempting to outline major interaction systems on the basis of information from different sources. The evidence concerning interactions that control development must come primarily from development itself. Both disturbed, as during regeneration and grafts, and normal development are relevant. Growing apices interact over relatively long distances. They reduce the development of similar apices and induce the development of axial tissues that connect them to the plant. Young shoot tissues also induce the development of root apices and vice versa. These various effects can be understood on the basis of a hormonal feedback involving auxins and cytokinins. Vascular differentiation, furthermore, is a cellular expression of these interactions. It occurs along the flow of auxin from the young tissues of the shoot towards the root apices. This flow is canalized by a positive feedback between cell polarization and the polar flow that both controls and results from this polarization. Structural relations between pholem and xylem, limitations of regeneration and the formation of rays all indicate the existence of additional, radial interactions. The fate of individual cells in development is varied and often follows no recognizable rules. This indicates interactions that operate on the size of a tissue or organ rather than on its precise cellular development. On the other hand, the continuity of plasmodesmata, wall thickenings and cytoplasmic strands demonstrate local interactions between neighbouring cells. It is concluded that, though the possibilities of developmental interactions may be bewildering, the list, when known, may not be all that long. The interactions specify orientation and quantity rather than the precise fate of cells. Their effects are gradual and they involve feedback loops. Growth factors, even known growth factors, play surprisingly large roles in cellular interactions. Such controls whose effect is gradual and general could provide a basis for developmental plasticity.

Cell Communication↗

Cytokinesis in Impatiens balsamina and the effect of caffeine.

A description of cytokinesis in cells of roots of Impatiens balsamina is given, and the effect of caffeine on this process. The disposition of organelles, microtubules and vesicles which form the new cell plate is similar in normal and caffeine treated roots. In sections cut in the plane of the developing cell plate, membranous arms radiating from common centres appear to play an important role in promoting efficient fusion of the vesicles, and caffeine reduces their occurrence. The possible presence of callose in the newly formed cell plate, and the formation of plasmodesmata, are discussed. Mechanisms of membrane fusion and effects of caffeine on this event are also considered.

Caffeine↗

Cell-to-cell communication in differentiation of mycelial fungi.

Electric and ionic interactions in fungal hyphae under controlled environmental conditions were characterized. The interactions were shown to be local and genetically determined. The possibility of their regulation by the blue-violet light and the membrane potential gradient between the adjacent cells was demonstrated. Septal pores providing the intercellular communication in fungi share common functional features with animal gap junctions, plant plasmodesmata, and microplasmodesmata of cyanobacteria. Integration of cells in a structurally and functionally heterogeneous ensemble takes place via local intercellular interactions due to a nonuniform distribution of ionic pumps and secondary transport systems in the apical and proximal hyphal cells. Cell differentiation during the hyphal growth results from the redistribution of functions in apical cells populations. The loss of polarity by growing intercalary hyphal fragments and equalization of ionic gradients during the regeneration of mechanically damaged intrahyphal fragments may indicate the existence of different growth mechanisms in the mycelium. Synchronization of functions of individual hyphal compartments exhibiting considerable electrophysiological heterogeneity takes place during the blue-light signal transduction, which triggers the hyphal differentiation processes.

Carotenoids↗

The endoplasmic reticulum of plant cells and its role in protein maturation and biogenesis of oil bodies.

The endoplasmic reticulum (ER) is the port of entry of proteins into the endomembrane system, and it is also involved in lipid biosynthesis and storage. This organelle contains a number of soluble and membrane-associated enzymes and molecular chaperones, which assist the folding and maturation of proteins and the deposition of lipid storage compounds. The regulation of translocation of proteins into the ER and their subsequent maturation within the organelle have been studied in detail in mammalian and yeast cells, and more recently also in plants. These studies showed that in general the functions of the ER in protein synthesis and maturation have been highly conserved between the different organisms. Yet, the ER of plants possesses some additional functions not found in mammalian and yeast cells. This compartment is involved in cell to cell communication via the plasmodesmata, and, in specialized cells, it serves as a storage site for proteins. The plant ER is also equipped with enzymes and structural proteins which are involved in the process of oil body biogenesis and lipid storage. In this review we discuss the components of the plant ER and their function in protein maturation and biogenesis of oil bodies. Due to the large number of cited papers, we were not able to cite all individual references and in many cases we refer the readers to reviews and references therein. We apologize to the authors whose references are not cited.

Endoplasmic Reticulum↗

Changes in macromolecular movement accompany organogenesis in thin cell layers of Torenia fournieri.

A range of fluorescently labelled probes of increasing molecular weight was used to monitor diffusion via the symplast in regenerating thin cell layer (TCL) explants of Torenia fournieri. An increase in intercellular movement of these molecules was associated with the earliest stages of vegetative shoot regeneration, with the movement of a 10 kDa dextran (FD 10000) observed between epidermal cells prior to the appearance of the first cell divisions. A low frequency of dextran movement in thin cell layers maintained under non-regenerating conditions was also observed, indicating a possible wound induced increase in intercellular movement. Dextran movement between epidermal cells reached a peak by day 4 of culture and then declined as cell division centres (CDCs) formed, became meristematic regions and finally emerged as adventitious shoots. Within CDCs, testing with small fluorescent probes (CF: carboxyfluorescein, mw 376 Da and F(Glu)3: fluorescein-triglutamic acid, mw 799 Da) revealed a mosaic of cell isolation and regions of maintained symplastic linkage. Within shoots, surface cells of the presumptive apical meristem permitted the intercellular movement of 10 kDa dextrans but epidermal cells of the surrounding leaf primordia did not permit dextran movement. In some cases, intercellular movement of CF was maintained within leaf primordia. Symplastic movement of labelled dextrans during regeneration in Torenia thin cell layers represents a significant increase in the basal size exclusion limit (SEL) of this tissue and reveals the potential for intercellular trafficking of developmentally related endogenous macromolecules.

Biological Transport↗