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  • 1
    Electronic Resource
    Electronic Resource
    Root contraction in hyacinth. II. Changes in tubulin levels, microtubule number and orientation associated with differential cell expansion (1988)
    Springer
    Planta 174 (1988), S. 446-452 
    Details
    ISSN: 1432-2048
    Keywords: Cell expansion ; Hyacinthus ; Microtubule ; Root contraction ; Tubulin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Root contraction in hyacinth (Hyacinthus orientalis L.) is marked by reoriented cell growth in the cortex of the contractile region. Cellular volume of the inner cortex enlarges fourfold during root contraction. This is associated with large increases in the radial and tangential dimensions and decreases in the longitudinal dimension of the cells. In order to determine the possible role of microtubules (MTs) in these changes we compared tubulin levels and MT numbers and orientation in contracted and non-contracted regions of hyacinth roots. Tubulin content was analysed by a radioimmunoassay; MT numbers and orientation were analyzed by counting profiles in sectioned material using transmission electron microscopy. Contracted tissue was found to have significantly higher levels of tubulin on a per-cell basis than non-contracted tissue, and also increased tubulin levels relative to total protein. The spatial MT frequencies were the same in contracted and non-contracted tissues, indicating a proportional increase in MT numbers in the expanded cells. Although the absolute spatial frequency of MTs was constant, the orientation, as determined by morphometric analysis of MT profiles, was not. While in the longitudinal section plane 42% of the MTs in the non-contracted cells were oblique, in the contracted cells the percentage of MTs presenting oblique profiles increased to 87%. Additionally, a qualitative difference in MTs was observed in contracted cells; electron-opaque material was seen peripherally associated with the MTs of the inner cortex. The changes in tubulin levels and in MT numbers as well as the qualitative differences in the MTs of contracted and non-contracted root regions indicate that, in hyacinth, reoriented cellular enlargement associated with root contraction cannot be explained simply by shifts in the arrangement of preexisting cortical MT arrays, but involves more complex changes in the cytoskeleton.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00634472
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  • 2
    Electronic Resource
    Electronic Resource
    Microtubule-binding proteins from carrot (1989)
    Springer
    Planta 177 (1989), S. 245-260 
    Details
    ISSN: 1432-2048
    Keywords: Cytoskeleton ; Daucus (microtubules) ; Microtubule-associated proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Microtubules (MTs) participate in several processes of fundamental importance to growth and development in higher plants, yet little is known about the proteins with which they associate. Information about these molecules is important because they probably play a role in mediating functional and structural differences between various MT arrays. As a first step in gaining insight into this problem, we have isolated, from suspension-cultured cells of carrot (Daucus carota L.), non-tubulin proteins which bind to and affect microtubules (MTs) in vitro. These proteins were isolated using taxol-stabilized neuronal MTs as an affinity substrate. They cause MT bundling at substoichiometric concentrations, support the assembly of tubulin in vitro, and at low concentrations, decorate single MTs in a periodic fashion. The bundled MTs formed in vitro share similarities with those seen in situ in a variety of plant cells, including a center-center spacing of 34 nm, cold stability, resistance to anti-microtubule drugs, and sensitivity to calcium. The bundling activity is specific; other cationic proteins, as well as poly-L-lysine, do not behave in a similar manner. The bundling activity is insensitive to ATP. By assaying bundling activity with dark-field microscopy and employing standard biochemical procedures, a small number of polypeptides involved in the bundling process were identified. Affinity-isolated antibodies to one of these polypeptides (Mr=76000) were found to co-localize with MTs in the cortical array of protoplasts. Our findings are discussed with reference to the importance of these proteins in the cell and to their relationship to microtubule-associated proteins in other eukaryotes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00392813
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  • 3
    Electronic Resource
    Electronic Resource
    Microtubule reorganization in tobacco BY-2 cells stably expressing GFP-MBD (2000)
    Springer
    Planta 210 (2000), S. 502-509 
    Details
    ISSN: 1432-2048
    Keywords: Key words: Copper-inducible promoter – Cytoskeleton – Green fluorescent protein (GFP) – Microtubule-associated protein (MAP4) – Microtubule –Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract.  Microtubule organization plays an important role in plant morphogenesis; however, little is known about how microtubule arrays transit from one organized state to another. The use of a genetically incorporated fluorescent marker would allow long-term observation of microtubule behavior in living cells. Here, we have characterized a Nicotiana tabacum L. cv. Bright Yellow 2 (BY-2) cell line that had been stably transformed with a gfp-mbd construct previously demonstrated to label microtubules (J. Marc et al., 1998, Plant Cell 10: 1927–1939). Fluorescence levels were low, but interphase and mitotic microtubule arrays, as well as the transitions between these arrays, could be observed in individual gfp-mbd-transformed cells. By comparing several attributes of transformed and untransformed cells it was concluded that the transgenic cells are not adversely affected by low-level expression of the transgene and that these cells will serve as a useful and accurate model system for observing microtubule reorganization in vivo. Indeed, some initial observations were made that are consistent with the involvement of motor proteins in the transition between the spindle and phragmoplast arrays. Our observations also support the role of the perinuclear region in nucleating microtubules at the end of cell division with a progressive shift of these microtubules and/or nucleating activity to the cortex to form the interphase cortical array.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004250050037
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  • 4
    Electronic Resource
    Electronic Resource
    The isolation, characterization and sequence of two divergent β-tubulin genes from soybean (Glycine max L.) (1987)
    Springer
    Plant molecular biology 10 (1987), S. 171-184 
    Details
    ISSN: 1573-5028
    Keywords: soybean β-tubulins ; genomic sequence ; genomic sequence ; protein sequence ; interspecies sequence comparison ; hydropathy analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two divergent β-tubulin genes (designated Sβ-1 and Sβ-2) were isolated by screening a soybean genomic library with a Chlamydomonas reinhardtii β-tubulin cDNA probe. Restriction fragment analysis of the clones recovered, and of soybean genomic DNA, indicated that these represent two unique classes of structurally different β-tubulin genes in the soybean genome. However, it is possible that unidentified members of these classes or additional highly divergent classes of β-tubulin genes (thus far undetected) exist in the soybean genome. The Sβ-1 and Sβ-2 genomic clones were sequenced, revealing that both are potentially functional genes which would encode β-tubulins of 445 and 449 amino acids, respectively. A comparison of their derived amino acid sequences with β-tubulins from several organisms showed that they are most homologous to Chlamydomonas β-tubulin (85–87%), with lesser degrees of homology to β-tubulins of vertebrate species (79–83%), Trypanosoma brucei (80–81%) and Saccharomyces cerevisiae (66–68%). The amino acid sequences of Sβ-1 and Sβ-2 are as divergent from each other as they are from the Chlamydomonas β-tubulin. The amino acids at the diverged positions in Sβ-2 are nearly all conservative substitutions while in Sβ-1, 18 of the 69 substitutions were non-conservative. Both soybean β-tubulin genes contain two introns in exactly the same positions. The first soybean intron is located in the same position as the third intron of the Chlamydomonas β-tubulin genes. Codon usage in the two soybean β-tubulins is remarkably similar (D 2=0.87), but differs from codon usage in other soybean genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00016154
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  • 5
    Electronic Resource
    Electronic Resource
    Characterization of a monoclonal antibody prepared against plant actin (1994)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 29 (1994), S. 339-344 
    Details
    ISSN: 0886-1544
    Keywords: microfilament ; phalloidin ; immunoblotting ; immunocytochemistry ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Anti-actin monoclonal antibodies were prepared using phalloidin-stabilized actin that was purified from pea roots by DNase I affinity chromatography. One monoclonal antibody, designated mAb3H11, bound plant actin in preliminary screenings and was further analyzed. Immunoblot analysis showed that this antibody had a high affinity for plant actin in crude and purified preparations but a low affinity for rabbit muscle actin. In immunoblots of plant extracts separated on two-dimensional gels it appeared to bind all actin isoforms recognized by the JLA20 anti-chicken actin antibody. Using immunofluorescent cytochemistry, the antibody was used to observe actin filaments in aldehyde-fixed and methanol-treated tobacco protoplasts. These results indicate that mAb3H11 should be a useful reagent for the study of plant actins. © 1994 Wiley-Liss, Inc.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970290406
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  • 6
    Electronic Resource
    Electronic Resource
    The expression of a chimeric soybean beta-tubulin gene in tobacco (1987)
    Springer
    Molecular genetics and genomics 209 (1987), S. 633-633 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00331178
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  • 7
    Electronic Resource
    Electronic Resource
    The expression of a chimeric soybean beta-tubulin gene in tobacco (1987)
    Springer
    Molecular genetics and genomics 207 (1987), S. 328-334 
    Details
    ISSN: 1617-4623
    Keywords: Tubulin ; Cell transformation ; Gene expression ; Ti plasmid ; Agrobacterium tumefaciens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A chimeric tubulin gene has been constructed by the fusion of a genomic sequence containing a truncated soybean beta-tubulin gene with 2 kb of upstream DNA to the 3′ untranslated region containing the polyadenylation signal from transcription unit 7 of the octopine Ti plasmid pGV117. The chimeric gene has been incorporated into the Ti plasmid transformation vector pGV3850::pAP2034, along with a selectable marker gene active in plants (the neomycin phosphotransferase II gene, conferring kanamycin resistance). Strains of Agrobacterium tumefaciens haboring the plasmids were used to transform Nicotiana tabacum by the leaf disk method and plants were regenerated from transformed cells. Transgenic plants were self fertilized and the segregation of kanamycin resistance was assayed. DNA and RNA were extracted from transgenic plants, fractionated by agarose gel electrophoresis, blotted to nitrocellulose and hybridized to a probe specific for the chimeric gene to assess its structure and expression in transgenic plants. The chimeric gene was stably integrated into the tobacco genome without rearrangements and it was expressed as a polyadenylated RNA of 1.7 kb in the transformants. Genetic analysis revealed that the kanamycin-resistant phenotype was inherited in a Mendelian fashion over two generations.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00331597
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