The cellular pathway of photosynthate transfer in the developing wheat grain. II. A structural analysis and histochemical studies of the pathway from the crease phloem to the endosperm cavity

In the developing wheat grain, photosynthate is transferred longitudinally along the crease phloem and then laterally into the endosperm cavity through the crease vascular parenchyma, pigment strand and nucellar projection. In order to clarify this cellular pathway of photosynthate unloading, and hence the controlling mechanism of grain filling, the potential for symplastic and apoplastic transfer was examined through structural and histochemical studies on these tissue types. It was found that cells in the crease region from the phloem to the nucellar projection are interconnected by numerous plasmodesmata and have dense cytoplasm with abundant mitochondria. Histochemical studies confirmed that, at the stage of grain development studied, an apoplastic barrier exists in the cell walls of the pigment strand. This barrier is composed of lignin, phenolics and suberin. The potential capacity for symplastic transfer, determined by measuring plasmodesmatal frequencies and computing potential sucrose fluxes through these plasmodesmata, indicated that there is sufficient plasmodesmatal cross-sectional area to support symplastic unloading of photosynthate at the rate required for normal grain growth. The potential capacity for membrane transport of sucrose to the apoplast was assessed by measuring plasma membrane surface areas of the various cell types and computing potential plasma membrane fluxes of sucrose. These fluxes indicated that the combined plasma membrane surface areas of the sieve element–companion cell (se–cc) complexes, vascular parenchyma and pigment strand are not sufficient to allow sucrose transfer to the apoplast at the observed rates. In contrast, the wall ingrowths of the transfer cells in the nucellar projection amplify the membrane surface area up to 22-fold, supporting the observed rates of sucrose transfer into the endosperm cavity. We conclude that photosynthate moves via the symplast from the se–cc complexes to the nucellar projection transfer cells, from where it is transferred across the plasma membrane into the endosperm cavity. The apoplastic barrier in the pigment strand is considered to restrict solute movement to the symplast and block apoplastic solute exchange between maternal and embryonic tissues. The implications of this cellular pathway in relation to the control of photosynthate transfer in the developing grain are discussed.     

Roles of external and cellular Cl− ions on the activation of an apical electrodiffusional Cl− pathway in toad skin

Summary This study is concerned with the short-circuit current,I _sc, responses of the Cl^–-transporting cells of toad skin submitted to sudden changes of the external Cl^– concentration. [Cl]₀. Sudden changes of [Cl]₀, carried out under apical membrane depolarization, allowed comparison of the roles of [Cl]₀ and [Cl]_cell on the activation of the apical Cl^– pathways. Equilibration of shortcircuited skins symmetrically in K-Ringer's solutions of different Cl^– concentrations permitted adjustment of [Cl]_cell to different levels. For a given Cl^– concentration (in the range of 11.7 to 117mm) on both sides of a depolarized apical membrane, this structure exhibits a high Cl^– permeability,P _(Cl)apical. On the other hand, for the same range of [Cl]_cell but with [Cl]₀=0,P _(Cl)apical is reduced to negligible values. These observations indicate that when the apical membrane is depolarizedP _(Cl)apical is modulated by [Cl]₀; in the absence of external Cl^– ions, intracellular Cl^– is not sufficient to activateP _(Cl)apical. Computer simulation shows that the fast Cl^– currents induced across the apical membrane by sudden shifts of [Cl]₀ from a control equilibrium value strictly follow the laws of electrodiffusion. For each experimental group, the computer-generatedI _sc versus ([Cl]_cell–[Cl]₀) curve which best fits the experimental data can only be obtained by a unique pair ofP _(Cl)apical andR _b (resistance of the basolateral membrane), thus allowing the calculation of these parameters. The electrodiffusional behavior of the net Cl^– flux across the apical membrane supports the channel nature of the apical Cl^– pathways in the Cl^–-transporting cell. Cl^– ions contribute significantly to the overall conductance of the basolateral membrane even in the presence of a high K concentration in the internal solution.     

LptB-LptF coupling mediates the closure of the substrate-binding cavity in the LptB2FGC transporter through a rigid-body mechanism to extract LPS

Lipopolysaccharides (LPS) are essential envelope components in many Gram-negative bacteria and provide intrinsic resistance to antibiotics. LPS molecules are synthesized in the inner membrane and then transported to the cell surface by the LPS transport (Lpt) machinery. In this system, the ATP-binding cassette (ABC) transporter LptB₂FGC extracts LPS from the inner membrane and places it onto a periplasmic protein bridge through a poorly understood mechanism. Here, we show that residue E86 of LptB is essential for coupling the function of this ATPase to that of its partners LptFG, specifically at the step where ATP binding drives the closure of the LptB dimer and the collapse of the LPS-binding cavity in LptFG that moves LPS to the Lpt periplasmic bridge. We also show that defects caused by changing residue E86 are suppressed by mutations altering either LPS structure or transmembrane helices in LptG. Furthermore, these suppressors also fix defects in the coupling helix of LptF, but not of LptG. Together, these results support a transport mechanism in which the ATP-driven movements of LptB and those of the substrate-binding cavity in LptFG are bi-directionally coordinated through the rigid-body coupling, with LptF’s coupling helix being important in coordinating cavity collapse with LptB dimerization.     

Cell sheet technology: a promising strategy in regenerative medicine

Regenerative medicine is a burgeoning field that is important to combat challenging diseases and functional impairments. Compared with traditional cell therapies with evident shortcomings (e.g., cell suspension injection or tissue engineering with scaffolds), scaffold-free cell sheet technology enables transplanted cells to be grafted and fully maintain their viability on target sites. Clinical and experimental studies have advanced the application of cell sheet technology to numerous tissues and organs (e.g., liver, cornea and bone). However, previous reviews have failed to discuss vital aspects of this rapidly developing technology, and many new challenges are gradually emerging. This review aims to provide a comprehensive introduction to cell sheet technology from cell selection to the ultimate applications of cell sheets, and challenges and future visions are also described.     

Centrosomes and cancer.

The centrosome functions as the major microtubule organizing center (MTOC) of the cell and as such it determines the number, polarity, and organization of interphase and mitotic microtubules. Cytoplasmic organization, cell polarity and the equal partition of chromosomes into daughter cells at the time of cell division are all dependent on the normal function of the centrosome and on its orderly duplication, once and only once, in each cell cycle. Malignant tumor cells show characteristic defects in cell and tissue architecture and in chromosome number that can be attributed to inappropriate centrosome behavior during tumor progression. In this review, we will summarize recent observations linking centrosome defects to disruption of normal cell and tissue organization and to chromosomal instability found in malignant tumors.     

Synthesis, characterization and ethylene polymerization activity of titanium aminopyridinato complexes

Three titanium complexes derived from 2-(2,6-difluoroanilino)pyridine and 2-(2-chloroanilino)pyridine were synthesized and characterized by X-ray diffraction or spectroscopic methods. All titanium complexes have been used to catalyze the polymerization of ethylene in the presence of MAO as cocatalyst. The mono(2,6-difluorophenylaminopyridinato) titanium catalyst was found to be more active in ethylene polymerization than the bis(2,6-difluorophenylaminopyridinato) and bis(2-chlorophenylaminopyridinato) titanium catalysts. ortho-Halogens disturbed the β-elimination transition state of ethylene polymerization and formed higher molar mass polyethylene than their unhalogenated congener. Due to fluxionality, the bis(2-chlorophenylaminopyridinato) titanium catalyst formed broader molar mass distribution than the bis(2,6-difluorophenylaminopyridinato) titanium catalysts.     

Phylogenetic and phenotypic structure among Banksia communities in south‐western Australia

Aim Phylogenetic and phenotypic patterns among coexisting banksias (Banksia, Proteaceae) in the infertile, fire‐prone landscapes of south‐western Australia were examined for evidence of community structuring. It was expected that closely related species would be spatially clustered (underdispersed) as a consequence of widespread recent speciation, strong edaphic fidelity and low dispersability. We also expected that edaphic filtering would result in phenotypic clustering of traits related to habitat specialization and that competitive exclusion among closely related species with similar regeneration biology and growth form would result in phenotypic overdispersion of these latter traits. Location Southwest Australian Floristic Region (SWAFR). Methods Based on published data for coexistence (richness and frequency) of Banksia species at 40 sites in the three floristic provinces, phylogenetic, soil type and morphological mean pairwise distance and mean nearest taxon distance were calculated for each site and compared with null communities. Patterns of co‐occurrence were examined at the local and subregional (provincial) scales. Results Of the 40 sites assessed, 21–30 displayed phylogenetic clustering of Banksia species (5–11 significantly) such that, overall, co‐occurring taxa were more closely related than expected by chance. Banksias in the Transitional Rainfall and Southeast Coastal Provinces were more likely to display phylogenetic clustering than in the High Rainfall Province. A significant trend for phylogenetic clustering associated with edaphic specialization (27–30 sites) was observed, as well as a significant trend for phenotypic overdispersion associated with growth form (25–28 sites). Results for regeneration biology depended on the metric used. Main conclusions We demonstrate spatial clustering of closely related banksias at the local and provincial scales, consistent with their restricted distribution (recent widespread speciation, patchy habitat availability and limited dispersability) in this geologically old and stable region. The clustering of closely related species may also be a consequence of habitat filtering linked to edaphic fidelity in the SWAFR flora, while overdispersion in growth form suggests that functional divergence favours coexistence in Banksia communities.     

Climate oscillations and species interactions: large‐scale congruence but regional differences in the phylogeographic structures of an alpine plant and its monophagous insect

Aim To predict the fate of alpine interactions involving specialized species, using a monophagous beetle and its host plant as a case study. Location The Alps. Methods We investigated genetic structuring of the herbivorous beetle Oreina gloriosa and its specific host‐plant Peucedanum ostruthium. We used genome fingerprinting (in the insect and the plant) and sequence data (in the insect) to compare the distribution of the main gene pools in the two associated species and to estimate divergence time in the insect, a proxy for the temporal origin of the interaction. We quantified the similarity in spatial genetic structures by performing a Procrustes analysis, a tool from shape theory. Finally, we simulated recolonization of an empty space analogous to the deglaciated Alps just after ice retreat by two lineages from two species showing unbalanced dependence, to examine how timing of the recolonization process, as well as dispersal capacities of associated species, could explain the observed pattern. Results Contrasting with expectations based on their asymmetrical dependence, patterns in the beetle and plant were congruent at a large scale. Exceptions occurred at a regional scale in areas of admixture, matching known suture zones in Alpine plants. Simulations using a lattice‐based model suggested these empirical patterns arose during or soon after recolonization, long after the estimated origin of the interaction c. 0.5 million years ago. Main conclusions Species‐specific interactions are scarce in alpine habitats because glacial cycles have limited the opportunities for co‐evolution. Their fate, however, remains uncertain under climate change. Here we show that whereas most dispersal routes are paralleled at a large scale, regional incongruence implies that the destinies of the species might differ under changing climate. This may be a consequence of the host dependence of the beetle, which locally limits the establishment of dispersing insects.     

Butterfly Response to Microclimatic Conditions Following Ponderosa Pine Restoration

Efforts to restore ponderosa pine ecosystems to open, park‐like conditions that predominated prior to European‐American settlement result in altered stand structure and increased landscape heterogeneity, potentially altering habitat suitability for invertebrates and other forest organisms. We examined the responses of two butterfly species, Colias eurytheme and Neophasia menapia, to microclimatic changes at structural edges created by experimental restoration treatments in northern Arizona. We monitored microclimate, including air temperature, light intensity, and vapor pressure deficit (VPD), on several mornings during butterfly releases. We placed adult butterflies at east‐ and west‐facing edges approximately one half‐hour before dawn to determine their behavioral response to microclimatic differences between east‐ and west‐facing edges. After sunrise, all three microclimatic variables were higher at east‐facing edges, and the difference in microclimate between the two edge orientations increased through early morning. For both species, butterflies placed at east‐facing edges flew earlier than butterflies at west‐facing edges. Colias eurytheme, an open‐habitat species, tended to move toward the treated forest during initial flight, while movements of Neophasia menapia, a forest‐dwelling species, did not differ from random flight. Our results indicate that butterflies respond to microclimatic factors associated with restoration treatments, while responses to structural changes in habitat vary among species, based on habitat and food plant preferences. These changes in forest structure and microclimate may affect the distribution of many mobile invertebrates in forested landscapes undergoing restoration treatments.