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  • 1
    ISSN: 1546-1718
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Microarrays provide the means for the large-scale analysis of gene expression patterns in living organisms. My laboratory is part of three federally funded projects that are directed toward an understanding of the regulation of gene expression in higher plants. In the first project, funded by the ...
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  • 2
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 444 (2006), S. 31-31 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Sir Michael Eisenstein, in his Technology Feature “Quality control” (〈weblink url="http://www.nature.com/nature/journal/v442/n7106/full/4421067a.html"〉Nature 442, 1067–1070; 2006), reports recent improvements in microarray technologies. He ...
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 60 (1984), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Conditions have been established for the rapid flow analysis of leaf protoplasts of Nicotiana tabacum L. cv. Xanthi using a flow cytometer-cell sorter. A procedure based upon chlorophyll autofluorescence was devised to permit the systematic evaluation of flow conditions in order to identify those under which protoplast damage was minimized. These conditions were employed for the flow sorting of protoplasts, following which it was possible to regenerate the sorted protoplasts into complete plants. The application of flow sorting is discussed for the rapid identification and selection of somatic hybrids produced by protoplast fusion.
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 51 (1981), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A microfluorimetric procedure, employing the fluorescent stain 33258 Hoechst, has been developed for the investigation of the process of DNA synthesis during the initial stages of culture of tobacco (N. tabacum cv. Xanthi) leaf protoplasts.In this system, the freshly-isolated protoplasts exhibited a unimodal distribution of nuclear DNA content characteristic of the diploid state. The almost immediate onset of DNA synthesis during culture resulted in a doubling of nuclear DNA levels prior to the first mitoses. Although the majority of the protoplasts subsequently entered into synchronous mitosis and cell division, a proportion of the remainder developed into large polyploid cells. Upon further culture, the polyploid cells became subdivided into clusters of small diploid cells. Measurement of total cell protein and cell volumes during culture indicated that a relationship existed between these parameters and the initiation of mitosis. The significance of these observations is discussed.
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 55 (1982), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The effect of 2,6-dichlorobenzonitrile (DB) and coumarin on tobacco protoplast development has been studied using a combination of flow microfluorimetry and quantitative fluorescence microscopy. The results indicated that, over the initial period of culture, DB largely inhibited cellulose production at the cell surface, but did not inhibit DNA synthesis or protein accumulation by the protoplasts. Continuous culture of protoplasts in DB resulted in the accumulation of an increased capacity for cellulose synthesis as expressed following the removal of inhibitor. The possible sites of the specific inhibitory action of DB are discussed.
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  • 6
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 53 (1981), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A microfluorimetric procedure, using Calcofluor White, has been developed for the measurement of cellulose biosynthesis by cultured protoplasts of tobacco (Nicotiana tabacum L. cv. Xanthi nc). The procedure was compared to a conventional method for cellulose estimation, that employing the anthrone reagent following exhaustive extraction of the developing cell walls. The results indicate that the amount of fluorescence emitted following Calcofluor White treatment is a specific measurement of cell wall cellulose levels. The procedure possesses the twin advantages of ease of manipulation and of greatly enhanced sensitivity (in the picogram range) as compared to other methods.
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  • 7
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] We have developed general techniques for the successful selection of plant heterokaryons from parental populations of plant protoplasts subjected to somatic fusion treatments. These techniques involve a combination of artificial fluorescence labeling, natural chlorophyll autofluorescence and ...
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Journal of chemical ecology 25 (1999), S. 1739-1757 
    ISSN: 1573-1561
    Keywords: Spinach ; Spinacia oleracea ; induced defense ; damage ; 20-hydroxyecdysone ; phytoecdysteroid ; methyl jasmonate ; jasmonic acid ; root herbivory ; Otiorhynchus sulcatus ; abscisic acid ; gibberellic acid ; indole-3-acetic acid ; 1-naphthaleneacetic acid ; trans-zeatin ; root–shoot ratio
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Spinach (Spinacia oleracea) foliage is known to synthesize and accumulate insect molting hormones, predominantly in the form of 20-hydroxyecdysone (20E). We previously demonstrated that root 20E accumulation is increased following root damage. We designed two further experiments to address root responses to both mechanical and insect damage. In plants grown hydroponically, removal of 35% or less of the root mass did not result in changes in root 20E levels. However, removal of 70% of the root mass stimulated 6.0- and 1.5-fold increases in the root and shoot 20E concentrations, respectively. The effects of insect damage on soil-grown plants were investigated by infesting plant roots with black vine weevil (BVW: Otiorhynchus sulcatus) larvae and allowing them to feed for seven days. Decreases in root mass occurred in young plants; however, no changes were detected in mature plants. In all cases, root herbivory resulted in at least a 3.0-fold increase in root 20E concentrations. Our previous experiments implicated jasmonic acid and the analog methyl jasmonate (MJ) in signaling the damage-induced accumulation of root 20E levels. We investigated the activity of other phytohormones and growth regulators (GRs) on the 20E accumulation patterns of young plants as a means of examining the significance of jasmonates in the induction response. Hydroponic additions of MJ (0.5 μM) and the synthetic auxin, 1-naphthaleneacetic acid (NAA; 0.5 μM), resulted in significant increases in root 20E levels. At the concentrations tested, indole-3-acetic acid (IAA), gibberellic acid (GA3), abscisic acid (ABA), and trans-zeatin (Z) had no effects on root 20E concentrations. However, both NAA (0.5–5.0 μM) and Z (5.0 μM) treatments caused increases in the root/shoot dry mass ratios, indicating shifts in resource allocation to the roots. Treatments involving ABA (5.0 μM) and Z (0.5–5.0 μM) caused significant increases in shoot 20E concentrations. No other hormone treatments altered shoot accumulation patterns. The mechanisms underlying the root 20E induction phenomena were investigated through the incorporation of [2-14C]mevalonic acid ([14C]MVA). Within one day, excised roots readily incorporated radioactivity into 20E from [14C]MVA. In intact plants, [14C]MVA absorbed by the roots was rapidly incorporated into root 20E pools following damage and MJ treatments. This implies that the wound-induced root 20E accumulation is the result of increased de novo 20E synthesis in the root.
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  • 9
    ISSN: 1432-2048
    Keywords: Nicotiana (transfection with VSVG) ; Protoplast (transfection) ; Plasma membrane ; Rhabdoviruses ; Protein (secretion, targeting) ; Vesicular stomatitis virus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Leaf protoplasts of tobacco (Nicotlana tabacum L.) were employed for transfection of chimeric transcriptional gene fusions comprising the 35S promoter from cauliflower mosaic virus, the coding sequence of the G-protein from vesicular stomatitis virus (VSVG) and the transcriptional terminator from the Agrobacterium tumefaciens nopaline-synthetase gene. Transient expression of the chimeric gene was monitored through Northern analysis of total protoplast RNA using a labeled VSV cDNA probe, and through Western-blot analysis of protoplast proteins using a polyclonal and-VSV antiserum. Although a single species of mRNA was detected in the transfected protoplasts, two glycoproteins differing in mass by approx. 9 kDa were detected by the antiserum. Biosynthesis of the VSVG isoforms was not impeded by chemical inhibitors of cell-wall production or of proline hydroxylation. Transfection using mutant forms of the VSVG coding sequence in which either one or both consensus glycosylation sites were removed resulted in the production of progressively smaller VSVG proteins. Those proteins produced from the double mutant had mobilities on sodium dodecyl sulfate-polyacrylamide gel electrophoresis that were very similar to those produced from the wild-type construct in the presence of tunicamycin. Analysis of protoplast homogenates by differential centrifugation showed that the two VSVG isoforms were exclusively associated with cellular membranes. The larger protein co-localized with the plasma membrane and with the organelles of the endomembrane-secretory pathway leading to the plasma membrane. The smaller protein was associated with membranes of lower isopycnic densities which were not identical to the endoplasmic reticulum. The larger protein displayed greater sensitivity than did the smaller to degradation in vivo by exogenously added protease. Immunofluorescence microscopy demonstrated that the VSVG isoforms were present both within the protoplasts and at the surface of the plasma membrane. The intracellular distribution was either punctate or reticulate. These results are consistent with the progressive and accurate glycosylation of the newly synthesized VSVG polypeptide during its passage through the endomembrane-secretory pathway, the access of the larger isoform to the cell surface, and the conversion of the larger to the small isoform by selective proteolysis.
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  • 10
    ISSN: 1573-5028
    Keywords: cloning ; methyltransferase ; phytosterol ; yeast complementation ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report the characterization of a higher-plant C-24 sterol methyltransferase by yeast complementation. A Zea mays endosperm expressed sequence tag (EST) was identified which, upon complete sequencing, showed 46% identity to the yeast C-24 methyltransferase gene (ERG6) and 75% and 37% amino acid identity to recently isolated higher-plant sterol methyltransferases from soybean and Arabidopsis, respectively. When placed under GAL4 regulation, the Z. mays cDNA functionally complemented the erg6 mutation, restoring ergosterol production and conferring resistance to cycloheximide. Complementation was both plasmid-dependent and galactose-inducible. The Z. mays cDNA clone contains an open reading frame encoding a 40 kDa protein containing motifs common to a large number of S-adenosyl-L-methionine methyltransferases (SMTs). Sequence comparisons and functional studies of the maize, soybean and Arabidopsis cDNAs indicates two types of C-24 SMTs exist in higher plants.
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