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Plant Cells (plant + cell)
Terms modified by Plant Cells Selected AbstractsThe characterization of novel mycorrhiza-specific phosphate transporters from Lycopersicon esculentum and Solanum tuberosum uncovers functional redundancy in symbiotic phosphate transport in solanaceous speciesTHE PLANT JOURNAL, Issue 2 2005Réka Nagy Summary Solanaceous species are among the >200 000 plant species worldwide forming a mycorrhiza, that is, a root living in symbiosis with soil-borne arbuscular-mycorrhizal (AM) fungi. An important parameter of this symbiosis, which is vital for ecosystem productivity, agriculture, and horticulture, is the transfer of phosphate (Pi) from the AM fungus to the plant, facilitated by plasma membrane-spanning Pi transporter proteins. The first mycorrhiza-specific plant Pi transporter to be identified, was StPT3 from potato [Nature414 (2004) 462]. Here, we describe novel Pi transporters from the solanaceous species tomato, LePT4, and its orthologue StPT4 from potato, both being members of the Pht1 family of plant Pi transporters. Phylogenetic tree analysis demonstrates clustering of both LePT4 and StPT4 with the mycorrhiza-specific Pi transporter from Medicago truncatula [Plant Cell, 14 (2002) 2413] and rice [Proc. Natl Acad. Sci. USA, 99 (2002) 13324], respectively, but not with StPT3, indicating that two non-orthologous mycorrhiza-responsive genes encoding Pi transporters are co-expressed in the Solanaceae. The cloned promoter regions from both genes, LePT4 and StPT4, exhibit a high degree of sequence identity and were shown to direct expression exclusively in colonized cells when fused to the GUS reporter gene, in accordance with the abundance of LePT4 and StPT4 transcripts in mycorrhized roots. Furthermore, extensive sequencing of StPT4 -like clones and subsequent expression analysis in potato and tomato revealed the presence of a close paralogue of StPT4 and LePT4, named StPT5 and LePT5, respectively, representing a third Pi transport system in solanaceous species which is upregulated upon AM fungal colonization of roots. Knock out of LePT4 in the tomato cv. MicroTom indicated considerable redundancy between LePT4 and other Pi transporters in tomato. [source] Celebrating Plant Cells: A Special Issue on Plant Cell BiologyJOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 8 2007Zhenbiao Yang [source] Mechanical Response of Single Plant Cells to Cell Poking: A Numerical Simulation ModelJOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 6 2006Rong Wang Abstract Cell poking is an experimental technique that is widely used to study the mechanical properties of plant cells. A full understanding of the mechanical responses of plant cells to poking force is helpful for experimental work. The aim of this study was to numerically investigate the stress distribution of the cell wall, cell turgor, and deformation of plant cells in response to applied poking force. Furthermore, the locations damaged during poking were analyzed. The model simulates cell poking, with the cell treated as a spherical, homogeneous, isotropic elastic membrane, filled with incompressible, highly viscous liquid. Equilibrium equations for the contact region and the non-contact regions were determined by using membrane theory. The boundary conditions and continuity conditions for the solution of the problem were found. The force-deformation curve, turgor pressure and tension of the cell wall under cell poking conditions were obtained. The tension of the cell wall circumference was larger than that of the meridian. In general, maximal stress occurred at the equator around. When cell deformation increased to a certain level, the tension at the poker tip exceeded that of the equator. Breakage of the cell wall may start from the equator or the poker tip, depending on the deformation. A nonlinear model is suitable for estimating turgor, stress, and stiffness, and numerical simulation is a powerful method for determining plant cell mechanical properties. (Managing editor: Wei Wang) [source] Sugar Metabolic Analysis of Suspensions of Plant Cells Using an FT-IR/ATR MethodBIOTECHNOLOGY PROGRESS, Issue 3 2001Atsushi Hashimoto A simple, rapid and accurate evaluation of the sugar uptake rate of suspended plant cells from culture media was developed with the predicted sugar contents measured by mid-infrared spectroscopy using a Fourier transform infrared (FT-IR) spectrometer equipped with an attenuated total reflectance (ATR) accessory. We performed plant cell cultivation with Nicotiana tabacum cv. Bright Yellow No.2 (TBY-2) in culture media, which had various combinations of glucose, fructose and sucrose concentrations at the initial stage, and measured simultaneously each sugar content in the medium by the FT-IR/ATR method. By applying a logistic function to the predicted sugar contents and cell density in the medium during cultivation, the specific sugar uptake rates by the suspended TBY-2 cells were easily and continuously obtained. Thus the kinetic sugar uptake phenomena by the TBY-2 cells were well confirmed overall using the developed method. Additionally it was found that the fraction of sucrose of the initial total sugar content might kinetically affect the sugar uptake process and cell growth. Also, the relationship between the nondimensional cell density and sucrose content could be classified into three groups on the basis of the initial fraction of sucrose. [source] Expression and Purification of Functional Human ,-1-Antitrypsin from Cultured Plant CellsBIOTECHNOLOGY PROGRESS, Issue 1 2001Jianmin Huang Human ,-1-antitrypsin (AAT), the most abundant protease inhibitor found in the blood, was expressed in rice embryonic tissue suspension cell culture. This was accomplished by cloning the codon-optimized AAT gene into a vector containing the rice RAmy3D promoter and its signal sequence. The synthetic gene incorporates codons synonymous with those found in highly expressed rice genes. Approximately 1000 stable transformed calli were produced by particle bombardment mediated transformation and were screened for high AAT expression using a porcine elastase inhibitory activity assay. The band shift assay also confirmed that rice-derived AAT is functional regarding its binding capability to the elastase substrate. Time course studies were conducted to determine the optimum, postinduction expression levels from cell culture. AAT expression equivalent to 20% of the total secreted proteins was achieved, and a purification scheme was developed that yielded active AAT with purity greater than 95%. The potential applications of purified plant-derived AAT for treatments of various AAT-deficient diseases are discussed. [source] Highly Chemoselective Reduction of Aromatic Nitro Compounds to the Corresponding Hydroxylamines Catalyzed by Plant Cells from a Grape (Vitis vinifera L.).CHEMINFORM, Issue 35 2005Feng Li Abstract For Abstract see ChemInform Abstract in Full Text. [source] Comparative Metabolism of ,- and ,- Peptides in the Insect Heliothis virescens and in Plant Cells of Black Mexican Sweet MaizeCHEMISTRY & BIODIVERSITY, Issue 9 2004Rob Lind The tripeptide H-Val-Ala-Leu-OH and the N -Ac-tetrapeptide amide Ac-Thr-Lys-Trp-Phe-NH2, and their , -peptidic counterparts H- ,3hVal- ,3hAla- ,3hLeu-OH and Ac- ,3hThr-(S),2hLys- ,3hTrp- ,3hPhe-NH2, respectively, have been injected into Heliothis virescens larvae and added to cell cultures of black mexican sweet maize. The body liquids of the larvae and the supernatant of the plant cell cultures were sampled 0, 2, 3, 6, 17, and/or 24,h after application and analyzed by LC/MS. While the two , -peptides were degraded rapidly in these environments, the concentration of the , -peptides was found to decrease very slowly. Thus, ca. 60% of the original amount of the , -tetrapeptide was detected in the liquids of the insect after 24,h. The plant cells did not seem to make use of the , -peptides at all, whereas, the , -tripeptide completely disappeared from the supernatant after 3,h. Thus, we have demonstrated, for the first time, the high stability of , -peptides against degradation and metabolism in an insect and a plant. Especially remarkable is the persistence of the , -tetrapeptide with its functionalised and, thus, ,metabolisable' side chains of Thr, Lys, Trp, and Phe in the insect larvae, which are known to have a high level of activity of oxidizing enzymes. The results described here match those of ADME investigations with radioactively labeled , -peptides in rats, where essentially complete stability has been observed, while environmental microorganisms have been found to biodegrade , -peptides, albeit slowly. Possible implications of these findings for biomedical and pest-control applications are proposed. [source] Plant cells and algae in bioreactorsENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 3 2009Atanas Pavlov Guest Editor No abstract is available for this article. [source] Functional genomics of phosphate antiport systems of plastidsPHYSIOLOGIA PLANTARUM, Issue 4 2003Ulf-Ingo Flügge Plant cells require a co-ordination of metabolism between their major compartments, the plastids and the cytosol, in particular as certain metabolic pathways are confined to either compartments. The inner envelope membrane of the plastids forms the major barrier for metabolite exchange and is the site for numerous transport proteins, which selectively catalyse metabolite exchanges characteristic for green and/or non-green tissues. This report is focused on the molecular biology, evolution and physiological function of the family of phosphate translocators (PT) from plastids. Until now, four distinct subfamilies have been identified and characterized, which all share inorganic phosphate as common substrate, but have different spectra of counter exchange substrates to fulfil the metabolic needs of individual cells and tissues. The PTs are named after their main transported substrate, triose phosphate (TPT), phosphoenolpyruvate (PPT), glucose 6-phosphate (GPT) and xylulose 5-P (XPT). All PTs belong to the TPT/nucleotide sugar transporter (NST) superfamily, which includes yet uncharacterized PT homologues from plants and other eukaryotes. Transgenic plants or mutants with altered transport activity of some of the PTs have been generated or isolated. The analysis of these plant lines revealed new insights in the co-ordination and flexibility of plant metabolism. [source] Trafficking of the human transferrin receptor in plant cells: effects of tyrphostin A23 and brefeldin ATHE PLANT JOURNAL, Issue 5 2006Elena Ortiz-Zapater Summary Plant cells possess much of the molecular machinery necessary for receptor-mediated endocytosis (RME), but this process still awaits detailed characterization. In order to identify a reliable and well-characterized marker to investigate RME in plant cells, we have expressed the human transferrin receptor (hTfR) in Arabidopsis protoplasts. We have found that hTfR is mainly found in endosomal (Ara7- and FM4-64-positive) compartments, but also at the plasma membrane, where it mediates binding and internalization of its natural ligand transferrin (Tfn). Cell surface expression of hTfR increases upon treatment with tyrphostin A23, which inhibits the interaction between the YTRF endocytosis signal in the hTfR cytosolic tail and the ,2-subunit of the AP2 complex. Indeed, tyrphostin A23 inhibits Tfn internalization and redistributes most of hTfR to the plasma membrane, suggesting that the endocytosis signal of hTfR is functional in Arabidopsis protoplasts. Co-immunoprecipitation experiments show that hTfR is able to interact with a , -adaptin subunit from Arabidopsis cytosol, a process that is blocked by tyrphostin A23. In contrast, treatment with brefeldin A, which inhibits recycling from endosomes back to the plasma membrane in plant cells, leads to the accumulation of Tfn and hTfR in larger patches inside the cell, reminiscent of BFA compartments. Therefore, hTfR has the same trafficking properties in Arabidopsis protoplasts as in animal cells, and cycles between the plasma membrane and endosomal compartments. The specific inhibition of Tfn/hTfR internalization and recycling by tyrphostin A23 and BFA, respectively, thus provide valuable molecular tools to characterize RME and the recycling pathway in plant cells. [source] Antagonistic Effects of Hydrogen Peroxide and Glutathione on Acclimation to Excess Excitation Energy in ArabidopsisIUBMB LIFE, Issue 1 2000Barbara Karpinska Abstract The redox status of the quinone B (QB) and plastoquinone (PQ) pools plays a key role in the cellular and systemic signalling processes that control acclimatory responses in plants. In this study, we demonstrate the effects of hydrogen peroxide and glutathione on acclimatory responses controlled by redox events in the proximity of the QB-PQ pools. Our results suggest that the chloroplast is a sink for H2O2 and that, paradoxically, high concentrations of H2O2 in the chloroplast protect the photosynthetic apparatus and the plant cell from photoinhibition and photooxidative damage. Excess glutathione, however, caused an effect antagonistic to that observed for high H2O2. An explanation of this apparent paradox and a hypothetical redox-signalling model are suggested. [source] Type III-dependent translocation of the Xanthomonas AvrBs3 protein into the plant cellMOLECULAR MICROBIOLOGY, Issue 1 2002Boris Szurek Summary Many plant pathogenic bacteria utilize a conserved type III secretion system (TTSS) to deliver effector proteins into the host tissue. Indirect evidence has suggested that at least some effector proteins are translocated from the bacterial cytoplasm into the plant cell. Using an immunocytochemical approach, we demonstrate that the type III effector AvrBs3 from Xanthomonas campestris pv. vesicatoria localizes to nuclei of infected pepper leaves. Importantly, AvrBs3 translocation was observed in situ in native tissues of susceptible and resistant plants. AvrBs3 was detected in the nucleus as soon as 4 h post infection, which was dependent on a functional TTSS and the putative translocator HrpF. N-terminal AvrBs3 deletion derivatives are no longer secreted by the TTSS in vitro and could not be detected inside the host cells, suggesting that the N-terminus of AvrBs3 is important for secretion. Deletion of the nuclear localization signals in the AvrBs3 C-terminus, which are required for the AvrBs3-mediated induction of the hypersensitive reaction in resistant pepper plants, abolished AvrBs3 localization to the nucleus. This is the first report on direct evidence for translocation of a native type III effector protein from a plant pathogenic bacterium into the host cell. [source] Identification of novel hrp -regulated genes through functional genomic analysis of the Pseudomonas syringae pv. tomato DC3000 genomeMOLECULAR MICROBIOLOGY, Issue 5 2002Julie Zwiesler-Vollick Summary Pseudomonas syringae pv. tomato ( Pst ) strain DC3000 infects the model plants Arabidopsis thaliana and tomato, causing disease symptoms characterized by necrotic lesions surrounded by chlorosis. One mechanism used by Pst DC3000 to infect host plants is the type III protein secretion system, which is thought to deliver multiple effector proteins to the plant cell. The exact number of type III effectors in Pst DC3000 or any other plant pathogenic bacterium is not known. All known type III effector genes of P. syringae are regulated by HrpS, an NtrC family protein, and the HrpL alternative sigma factor, which presumably binds to a conserved cis element (called the ,hrp box') in the promoters of type III secretion-associated genes. In this study, we designed a search motif based on the promoter sequences conserved in 12 published hrp operons and putative effector genes in Pst DC3000. Seventy-three predicted genes were retrieved from the January 2001 release of the Pst DC3000 genome sequence, which had 95% genome coverage. The expression of the 73 genes was analysed by microarray and Northern blotting, revealing 24 genes/operons (including eight novel genes), the expression of which was consistently higher in hrp -inducing minimal medium than in nutrient-rich Luria,Bertani broth. Expression of all eight genes was dependent on the hrpS gene. Most were also dependent on the hrpL gene, but at least one was dependent on the hrpS gene, but not on the hrpL gene. An AvrRpt2-based type III translocation assay provides evidence that some of the hrpS -regulated novel genes encode putative effector proteins. [source] pH regulation in an acidophilic green alga , a quantitative analysisNEW PHYTOLOGIST, Issue 2 2009Birgit Bethmann Summary ,,Short-term cytosolic pH regulation has three components: H+ binding by buffering groups; H+ transport out of the cytosol; and H+ transport into the vacuole. To understand the large differences plants show in their tolerance to acidic environments, these three components were quantified in the acidophilic unicellular green alga Eremosphaera viridis. ,,Intracellular pH was recorded using ion-selective microelectrodes, whereas constant doses of weak acid were applied over different time intervals. A mathematical model was developed that describes the recorded cytosolic pH changes. Recordings of cytosolic K+ and Na+ activities, and application of anion channel inhibitors, revealed which ion fluxes electrically compensate H+ transport. ,,The cytosolic buffer capacity was , = 30 mM. Acidification resulted in a substantial and constant H+ efflux that was probably driven by the plasmalemma H+ -ATPase, and a proportional pH regulation caused by H+ pumped into the vacuole. Under severe cytosolic acidification (, 1 pH) more than 50% of the ATP synthesized was used for H+ pumping. While H+ influx into the vacuole was compensated by cation release, H+ efflux out of the cell was compensated by anion efflux. ,,The data presented here give a complete and quantitative picture of the ion fluxes during acid loading in an acidophilic green plant cell. [source] Cellular mechanisms of potassium transport in plantsPHYSIOLOGIA PLANTARUM, Issue 4 2008Dev T. Britto Potassium (K+) is the most abundant ion in the plant cell and is required for a wide array of functions, ranging from the maintenance of electrical potential gradients across cell membranes, to the generation of turgor, to the activation of numerous enzymes. The majority of these functions depend more or less directly upon the activities and regulation of membrane-bound K+ transport proteins, operating over a wide range of K+ concentrations. Here, we review the physiological aspects of potassium transport systems in the plasma membrane, re-examining fundamental problems in the field such as the distinctions between high- and low-affinity transport systems, the interactions between K+ and other ions such as NH4+ and Na+, the regulation of cellular K+ pools, the generation of electrical potentials and the problems involved in measurement of unidirectional K+ fluxes. We place these discussions in the context of recent discoveries in the molecular biology of K+ acquisition and produce an overview of gene families encoding K+ transporters. [source] Fluorescent protein fusions to a human immunodeficiency virus monoclonal antibody reveal its intracellular transport through the plant endomembrane systemPLANT BIOTECHNOLOGY JOURNAL, Issue 7 2008Sarah L. Irons Summary In order to further understand the production and intracellular trafficking of pharmaceutical proteins in plants, the light and heavy chains (LC and HC) of the human immunodeficiency virus neutralizing monoclonal antibody 2G12 were fused to fluorescent proteins [Venus and monomeric red fluorescent protein (mRFP)] to enable the visualization of their passage through the plant cell. Co-expression of LC and HC with various markers of the endomembrane system demonstrated that LC fusions were found in mobile punctate structures, which are likely to be pre-vacuolar compartments (PVCs) as a proportion of the LC fusions were found to be located in the vacuole. In addition, apoplast labelling was also observed with a 2G12LC-RFP fusion. The HC fusion expressed alone was found only in the endoplasmic reticulum (ER). When the LC and HC fusions were expressed together, they were found to co-locate to larger punctate structures, which were morphologically distinct from any observed on expression of LC or HC alone. These structures appeared to be in close association with the ER and their labelling partially overlapped with PVC marker fluorescence, but no increase in apoplast labelling was observed. Co-immunoprecipitation data demonstrated that the presence of the fluorescent proteins did not affect the assembly of the antibody, and also showed the association of BiP with the antibody chains. The antigen-binding activity of the Venus-fused 2G12 antibody was confirmed by enzyme-linked immunosorbent assay. [source] A recombinant multimeric immunoglobulin expressed in rice shows assembly-dependent subcellular localization in endosperm cellsPLANT BIOTECHNOLOGY JOURNAL, Issue 1 2005Liz Nicholson Summary To investigate the role of subunit assembly in the intracellular deposition of multimeric recombinant proteins, we expressed a partially humanized secretory immunoglobulin in rice endosperm cells and determined the subcellular locations of the assembled protein and its individual components. Transgenic rice plants expressing either individual subunits or all the subunits of the antibody were generated by particle bombardment, and protein localization was determined by immunoelectron microscopy. Assembly of the antibody was confirmed by immunoassay and coimmunoprecipitation. Immunolocalization experiments showed no evidence for secretion of the antibody or any of its components to the apoplast. Rather, the nonassembled light chain, heavy chain and secretory component accumulated predominantly within endoplasmic reticulum-derived protein bodies, while the assembled antibody, with antigen-binding function, accumulated specifically in protein storage vacuoles. These results show that the destination of a complex recombinant protein within the plant cell is influenced by its state of assembly. [source] Type III effectors orchestrate a complex interplay between transcriptional networks to modify basal defence responses during pathogenesis and resistanceTHE PLANT JOURNAL, Issue 1 2006William Truman Summary To successfully infect a plant, bacterial pathogens inject a collection of Type III effector proteins (TTEs) directly into the plant cell that function to overcome basal defences and redirect host metabolism for nutrition and growth. We examined (i) the transcriptional dynamics of basal defence responses between Arabidopsis thaliana and Pseudomonas syringae and (ii) how basal defence is subsequently modulated by virulence factors during compatible interactions. A set of 96 genes displaying an early, sustained induction during basal defence was identified. These were also universally co-regulated following other bacterial basal resistance and non-host responses or following elicitor challenges. Eight hundred and eighty genes were conservatively identified as being modulated by TTEs within 12 h post-inoculation (hpi), 20% of which represented transcripts previously induced by the bacteria at 2 hpi. Significant over-representation of co-regulated transcripts encoding leucine rich repeat receptor proteins and protein phosphatases were, respectively, suppressed and induced 12 hpi. These data support a model in which the pathogen avoids detection through diminution of extracellular receptors and attenuation of kinase signalling pathways. Transcripts associated with several metabolic pathways, particularly plastid based primary carbon metabolism, pigment biosynthesis and aromatic amino acid metabolism, were significantly modified by the bacterial challenge at 12 hpi. Superimposed upon this basal response, virulence factors (most likely TTEs) targeted genes involved in phenylpropanoid biosynthesis, consistent with the abrogation of lignin deposition and other wall modifications likely to restrict the passage of nutrients and water to the invading bacteria. In contrast, some pathways associated with stress tolerance are transcriptionally induced at 12 hpi by TTEs. [source] Optimal nitrogen supply as a key to increased and sustained production of a monoclonal full-size antibody in BY-2 suspension culture,BIOTECHNOLOGY & BIOENGINEERING, Issue 2 2010T. Holland Abstract Plant cell cultures have been used as expression hosts for recombinant proteins for over two decades. The quality of plant cell culture-produced proteins such as full-size monoclonal antibodies has been shown to be excellent in terms of protein folding and binding activity, but the productivity and yield fell short of what was achieved using mammalian cell culture, in which the key to gram-per-liter expression levels was strain selection and medium/process optimization. We carried out an extensive media analysis and optimization for the production of the full-size human anti-HIV antibody 2G12 in N. tabacum cv. BY-2. Nitrogen source and availability was found to be one key factor for the volumetric productivity of plant cell cultures. Increased amounts of nitrate in the culture medium had a dramatic impact on protein yields, resulting in a 10,20-fold increase in product accumulation through a combination of enhanced secretion and higher stability. The results were scalable from shake flasks to stirred-tank bioreactors, where the maximum yield per cultivation volume was 8,mg,L,1 over 7 days. During the stationary phase, antibody levels were 150-fold higher in nitrogen-enriched medium compared to standard medium. The enhanced medium appeared not to affect antibody quality and activity, as determined by Western blots, surface plasmon resonance binding assays and N -glycan analysis. Biotechnol. Bioeng. 2010;107: 278,289. © 2010 Wiley Periodicals, Inc. [source] Manufacturing antibodies in the plant cellBIOTECHNOLOGY JOURNAL, Issue 12 2009Diego Orzáez Dr. Abstract Plants have long been considered advantageous platforms for large-scale production of antibodies due to their low cost, scalability, and the low chances of pathogen contamination. Much effort has therefore been devoted to efficiently producing mAbs (from nanobodies to secretory antibodies) in plant cells. Several technical difficulties have been encountered and are being overcome. Improvements in production levels have been achieved by manipulation of gene expression and, more efficiently, of cell targeting and protein folding and assembly. Differences in mAb glycosylation patterns between animal and plant cells are being successfully addressed by the elimination and introduction of the appropriate enzyme activities in plant cells. Another relevant battlefield is the dichotomy between production capacity and speed. Classically, stably transformed plant lines have been proposed for large scale mAb production, whereas the use of transient expression systems has always provided production speed at the cost of scalability. However, recent advances in transient expression techniques have brought impressive yield improvements, turning speed and scalability into highly compatible assets. In the era of personalized medicines, the combination of yield and speed, and the advances in glyco-engineering have made the plant cell a serious contender in the field of recombinant antibody production. [source] Development of Auxotrophic Agrobacterium tumefaciens for Gene Transfer in Plant Tissue CultureBIOTECHNOLOGY PROGRESS, Issue 3 2004Jason I. Collens Auxotrophic strains of Agrobacterium tumefaciens were generated for use in liquid co-culture with plant tissue for transient gene expression. Twenty-one auxotrophs were recovered from 1,900 tetracycline-resistant insertional mutants generated with a suicide vector transposon mutagenesis system. Twelve of these auxotrophs were characterized on a nutrient matrix. Isolates were screened for growth in plant cell and root culture, and three auxotrophs were identified that had limited growth: adenine (ade-24), leucine (leu-27), and cysteine (cys-32). Ade-24 displayed poor T-DNA delivery in a transient expression test delivering GUS from a binary vector, while cys-32 displayed the best ability to deliver DNA of these three auxotrophs. The growth yield of cys-32 on cysteine was assessed to provide a quantitative basis for co-culture nutrient supplementation. The utility of cys-32 for delivering T-DNA to plant tissues is demonstrated, where an 85-fold enhancement in GUS expression over wild-type A. tumefacienswas achieved. [source] Manipulation of Intracellular Auxin in a Single Cell by Light with Esterase-Resistant Caged AuxinsCHEMBIOCHEM, Issue 13 2009Naoyuki Kusaka Abstract Auxin, a plant hormone, is polar transported from its site of production. This auxin polar transport system establishes an auxin gradient in plant tissue that is necessary for proper plant development. Therefore, the spatial effect of the auxin gradient on plant development is highly important for the understanding of plant auxin responses. Herein we report the design, syntheses and biological properties of esterase-resistant caged auxins. The conventional caging group, 2-nitrobenzyl ester, was found to be enzymatically hydrolyzed in plant cells and released original auxin without photolysis. The esterase-resistant caging group, (2,5-dimethoxyphenyl)(2-nitrobenzyl) ester, (DMPNB) was designed to improve the stability of caged auxins. Three auxins, indole 3-acetic acid, naphthalene 1-acetic acid and 2,4-dichlorophenoxy acetic acid were caged with the DMPNB caging group. DMPNB-caged auxins were inactive within a plant cell until photolysis, but they release auxins with photoirradiation to activate auxin-responsive gene expression. We demonstrated spatial and temporal control of intracellular auxin levels with photoirradiation by using this caged auxin system and were able to photocontrol the physiological auxin response in Arabidopsis plants. Additionally, the photoirradiation of DMPNB-caged auxin within a single cell can manipulate the intracellular auxin level and triggers auxin response. [source] Microtubule-dependent motility and orientation of the cortical endoplasmic reticulum in elongating characean internodal cellsCYTOSKELETON, Issue 3 2009Ilse Foissner Abstract Motility of the endoplasmic reticulum (ER) is predominantly microtubule- dependent in animal cells but thought to be entirely actomyosin-dependent in plant cells. Using live cell imaging and transmission electron microscopy to examine ER motility and structural organization in giant internodal cells of characean algae, we discovered that at the onset of cell elongation, the cortical ER situated near the plasma membrane formed a tight meshwork of predominantly transverse ER tubules that frequently coaligned with microtubules. Microtubule depolymerization increased mesh size and decreased the dynamics of the cortical ER. In contrast, perturbing the cortical actin array with cytochalasins did not affect the transverse orientation but decreased mesh size and increased ER dynamics. Our data suggest that myosin-dependent ER motility is confined to the ER strands in the streaming endoplasm, while the more sedate cortical ER uses microtubule-based mechanisms for organization and motility during early stages of cell elongation. We show further that the ER has an inherent, NEM-sensitive dynamics which can be altered via interaction with the cytoskeleton and that tubule formation and fusion events are cytoskeleton-independent. Cell Motil. Cytoskeleton 2009. © 2009 Wiley-Liss, Inc. [source] Potential impacts of climate change on Sub-Saharan African plant priority area selectionDIVERSITY AND DISTRIBUTIONS, Issue 6 2006Colin J. McClean ABSTRACT The Global Strategy for Plant Conservation (GSPC) aims to protect 50% of the most important areas for plant diversity by 2010. This study selects sets of 1-degree grid cells for 37 sub-Saharan African countries on the basis of a large database of plant species distributions. We use two reserve selection algorithms that attempt to satisfy two of the criteria set by the GSPC. The grid cells selected as important plant cells (IPCs) are compared between algorithms and in terms of country and continental rankings between cells. The conservation value of the selected grid cells are then considered in relation to their future species complement given the predicted climate change in three future periods (2025, 2055, and 2085). This analysis uses predicted climate suitability for individual species from a previous modelling exercise. We find that a country-by-country conservation approach is suitable for capturing most, but not all, continentally IPCs. The complementarity-based reserve selection algorithms suggest conservation of a similar set of grid cells, suggesting that areas of high plant diversity and rarity may be well protected by a single pattern of conservation activity. Although climatic conditions are predicted to deteriorate for many species under predicted climate change, the cells selected by the algorithms are less affected by climate change predictions than non-selected cells. For the plant species that maintain areas of climatic suitability in the future, the selected set will include cells with climate that is highly suitable for the species in the future. The selected cells are also predicted to conserve a large proportion of the species richness remaining across the continent under climate change, despite the network of cells being less optimal in terms of future predicted distributions. Limitations to the modelling are discussed in relation to the policy implications for those implementing the GSPC. [source] Determination of amino acids by micellar EKC: Recent advances in method development and novel applications to different matricesELECTROPHORESIS, Issue 1 2008Paolo Iadarola Professor Abstract The extensive use of CE for the analysis of amino acids has been well documented in a series of research articles and reviews. Aim of this report is to address the attention of the reader on the recent advances of micellar electrokinetic chromatography for the separation and determination of these analytes. Enhancements in selectivity of this technique through the use of pseudostationary phases containing mixed micelles, polymers, and chiral selectors are presented. Selected applications concerning separation and quantitation of even minute amounts of amino acids in: (i) biological fluids; (ii) microdialysates; (iii) plant cells; (iv) food stuff; and (v) pharmaceutical formulations have also been covered. Advantages of MEKC over other techniques for the amino acid analysis have been underlined. [source] Intracellular FITC-derivatization with PEGELECTROPHORESIS, Issue 21 2005Fanguo Chen Abstract In order to investigate the amino acids (AAs) in plant cells, we explore an avenue for intracellular derivatization with FITC. In this method, FITC was used to mark AAs in living protoplasts derived from embryogenic calli of common wheat (Triticum aestivum L.,c.v. Jinan,177) mediated by PEG. After FITC-derivatization, the AAs in the lysate were determined by CE. The result reveals that this PEG method can be used to transfer FITC into plant cells efficiently, which provides a good method for AA,analysis in plant cells. [source] Plants, gall midges, and fungi: a three-component systemENTOMOLOGIA EXPERIMENTALIS ET APPLICATA, Issue 1 2008Odette Rohfritsch Abstract Larvae of gall midges (Diptera: Cecidomyiidae) induce the activation of plant cells, partial cell lysis, and differentiation of nutritive tissue. Specialized nutritive tissue is essential for larval development and plays a key role in gall organization. Midges of the tribes Lasiopterini and Asphondyliini, however, do not induce nutritive tissues as part of the formation of their galls. Instead, these ,ambrosia galls' contain fungal mycelia that line the interior surface of the chambers. The fungi not only provide Lasiopterini with nutrition, they also penetrate the stems, induce the lysis of the middle lamella of host cells, and open a channel to the vascular bundles. Larvae of Lasioptera arundinis (Schiner) (Lasiopterini) follow the fungus and feed on its mycelium along with adjoining stem cells of Phragmites australis (Cav.) Trin. (Poaceae). Eggs together with fungal conidia are deposited by the imago on the host. Asphondyliini use a needle-like ovipositor to introduce fungal conidia and eggs into the organs they attack. Larvae of Schizomyia galiorum Kieffer (Asphondyliini) are unable to initiate the gall or to develop in the flowers of Galium mollugo L. (Rubiaceae) without their fungal associate. In this article, I provide an overview of oviposition behaviour in the Asphondyliini, as well as descriptions of the ovipositor and the female post-abdominal segments. Gall formation by Lasiopterini and Asphondyliini and the role of associated fungi are discussed, as is the role of the fungus as an inquiline or an organizer of gall tissues and a nutritive device. [source] Avirulence proteins from haustoria-forming pathogensFEMS MICROBIOLOGY LETTERS, Issue 2 2007Ann-Maree Catanzariti Abstract A major insight that has emerged in the study of haustoria-forming plant pathogens over the last few years is that these eukaryotic biotrophs deliver suites of secreted proteins into host cells during infection. This insight has largely derived from successful efforts to identify avirulence (Avr) genes and their products from these pathogens. These Avr genes, identified from a rust and a powdery mildew fungus and three oomycete species, encode small proteins that are recognized by resistance proteins in the host plant cytoplasm, suggesting that they are transported inside plant cells during infection. These Avr proteins probably represent examples of fungal and oomycete effector proteins with important roles in subverting host cell biology during infection. In this respect, they represent a new opportunity to understand the basis of disease caused by these biotrophic pathogens. Elucidating how these pathogen proteins gain entry into plant cells and their biological function will be key questions for future research. [source] Study of the mechanism of microwave-assisted extraction of Mahonia bealei (Fort.) leaves and Chrysanthemum morifolium (Ramat.) petalsFLAVOUR AND FRAGRANCE JOURNAL, Issue 3 2004Shan Gao Abstract A study of microwave-assisted extraction (MAE) for berberine in Mahonia bealei (Fort.) was carried out with batch equipment, in order to investigate the mechanism of the extraction related to structural changes in the glands. The extracts were analysed by ultraviolet-visible spectrophotometry at 347 nm. The parameters investigated were solvent types, the intensity of microwave energy and the process ratio (g/ml) of materials to solvent volume. The microwave-assisted extraction of different moisture content of materials was developed and optimized by means of three-factor and three-level orthogonal designs. Electron and optical micrographs of M. bealei (Fort.) leaves and Chrysanthemum morifolium (Ramat.) petals showed that the mechanism of the extractions was related to structural changes in the plant cells. Copyright © 2004 John Wiley & Sons, Ltd. [source] Functional analysis of the sea urchin-derived arylsulfatase (Ars)-element in mammalian cellsGENES TO CELLS, Issue 9 2006Satoshi Watanabe An insulator is a DNA sequence that has both enhancer-blocking activity, through its ability to modify the influence of neighboring cis -acting elements, and a barrier function that protects a transgene from being silenced by surrounding chromatin. Previously, we isolated and characterized a 582-bp-long element from the sea urchin arylsulfatase gene (Ars). This Ars -element was effective in sea urchin and Drosophila embryos and in plant cells. To investigate Ars -element activity in mammalian cells, we placed the element between the cytomegalovirus enhancer and a luciferase (luc) expression cassette. In contrast to controls lacking the Ars -element, NIH3T3 and 293T cells transfected with the element-containing construct displayed reduced luciferase activities. The Ars -element therefore acts as an enhancer-blocking element in mammalian cells. We assessed the barrier activity of the Ars -element using vectors in which a luc expression cassette was placed between two elements. Transfection experiments demonstrated that luc activity in these vectors was approximately ten-fold higher than in vectors lacking elements. Luc activities were well maintained even after 12 weeks in culture. Our observations demonstrate that the Ars -element has also a barrier activity. These results indicated that the Ars -element act as an insulator in mammalian cells. [source] | |||||||||||||||||||||||||||||||||||||