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  • 1
    Electronic Resource
    Electronic Resource
    Analysis of carbohydrate metabolism of CPD antisense plants and the brassinosteroid-deficient cbb1 mutant (2002)
    Oxford, UK : Blackwell Science, Ltd
    Plant, cell & environment 25 (2002), S. 0 
    Details
    ISSN: 1365-3040
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Brassinosteroids (BRs) are essential regulators of growth and development. BR-deficient mutants such as cpd/cbb3 and dwf4 display extreme dwarfism due to a failure in cell elongation. To avoid the severe pleiotropic effects caused by the extreme growth defect, transgenic Arabidopsis lines carrying a construct for antisense inhibition of CPD gene expression were established and subjected to physiological analysis. The CPD-antisense (α-CPD) lines display characteristic phenotypic alterations of BR-deficient plants such as reduced stem and petiole growth, smaller leaves, and a slightly delayed development. The observed changes are intermediate between the corresponding loss-of-function mutant (cbb3) and wild-type plants. In the present study, the primary carbon metabolism of the transgenic lines as well as the BR-deficient cbb1 (dwf1-6/dim) mutant was analysed. Gas exchange measurements indicated a reduced assimilatory capacity of the α-CPD plants. Soil-grown α-CPD as well as cbb1 (dwf1-6) mutant plants show a clear reduction in starch content. The metabolic alterations are accompanied by altered enzyme activities such as reduced invertase and cytosolic β-amylase activity, and altered expression patterns of genes such as Atbfruct1, Asus1, and ct-Bmy (encoding a cell wall invertase, sucrose synthase, and plastidic β-amylase, respectively). The impaired carbon assimilation, as well as the altered enzyme activities and gene expression patterns in the α-CPD and cbb1 (dwf1-6) plants, demonstrate the necessity of normal CPD and DIM expression for proper carbon uptake and metabolism and may point to an essential function of BRs. The impaired growth of BR-deficient plants may be (at least in part) due to reduced photosynthesis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1046/j.1365-3040.2002.00860.x
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  • 2
    Electronic Resource
    Electronic Resource
    Molecular cloning and expression analyses of mitochondrial and plastidic isoforms of cysteine synthase (O-acetylserine(thiol)lyase) fromArabidopsis thaliana (1999)
    Springer
    Amino acids 16 (1999), S. 113-131 
    Details
    ISSN: 1438-2199
    Keywords: Amino acids ; Targeting ; Mitochondria ; Chloroplasts ; Cysteine synthase ; Transit peptide ; Transgenic plants ; Processing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Cysteme synthase, the key enzyme for fixation of inorganic sulfide, catalyses the formation of cysteine from O-acetylserine and inorganic sulfide. Here we report the cloning of cDNAs encoding cysteine synthase isoforms fromArabidopsis thaliana. The isolated cDNA clones encode for a mitochondrial and a plastidic isoform of cysteine synthase (O-acetylserine (thiol)-lyase, EC 4.2.99.8), designated cysteine synthase C (AtCS-C, CSase C) and B (AtCS-B; CSase B), respectively.AtCS-C andAtCS-B, having lengths of 1569-bp and 1421-bp, respectively, encode polypeptides of 430 amino acids (∼45.8 kD) and of 392 amino acids (∼ 41.8 kD), respectively. The deduced amino acid sequences of the mitochondrial and plastidic isoforms exhibit high homology even with respect to the presequences. The predicted presequence of AtCS-C has a N-terminal extension of 33 amino acids when compared to the plastidic isoform. Northern blot analysis showed thatAtCS-C is higher expressed in roots than in leaves whereas the expression ofAtCS-B is stronger in leaves. Furthermore, gene expression of both genes was enhanced by sulfur limitation which in turn led to an increase in enzyme activity in crude extracts of plants. Expression of theAtCS-B gene is regulated by light. The mitochondrial, plastidic and cytosolic (Hesse and Altmann, 1995) isoforms of cysteine synthase ofArabidopsis are able to complement a cysteine synthasedeficient mutant ofEscherichia coli unable to grow on minimal medium without cysteine, indicating synthesis of functional plant proteins in the bacterium. Two lines of evidence proved thatAtCS-C encodes a mitochondrial form of cysteine synthase; first, import ofin vitro translation products derived from AtCS-C in isolated intact mitochondria and second, Western blot analysis of mitochondria isolated from transgenic tobacco plants expressing AtCS-C cDNA/c-myc DNA fusion protein.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01321531
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  • 3
    Electronic Resource
    Electronic Resource
    Establishment of a gene tagging system in Arabidopsis thaliana based on the maize transposable element Ac (1992)
    Springer
    Theoretical and applied genetics 84 (1992), S. 371-383 
    Details
    ISSN: 1432-2242
    Keywords: Arabidopsis thaliana ; Ds transposition ; Transposon tagging ; Insertion mutagenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An Ac-derived, two-component transposable element system has been developed and analyzed with respect to its use in Arabidopsis thaliana. This system consists of an immobilized Ac element (“Ac clipped wing”, Accl) as the source of transactivating transposase and a nonautonomous “Ds” element, DsA, which is inserted into a chimaeric neomycinphosphotransferase gene used as excision marker. After separate introduction of Acc1 and DsA into Arabidopsis thaliana, progeny analysis of crosses between five different Accl lines and seven different DsA lines shows that: (1) different Accl lines differ greatly in their capacity to transactivate DsA; (2) different DsA lines do not differ significantly with respect to DsA transactivation by one Accl line; (3) reintegration of excised DsA elements, both at (genetically) linked and unlinked sites, occurs in about 50% of the excision events; and (4) plants with a high rate of somatic excisions can be used as source of new DsA transpositions, allowing the creation of a large number of independent DsA insertions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00229496
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  • 4
    Electronic Resource
    Electronic Resource
    Selection for enhanced germinal excision of Ac in transgenic Arabidopsis thaliana (1993)
    Springer
    Theoretical and applied genetics 86 (1993), S. 919-926 
    Details
    ISSN: 1432-2242
    Keywords: Ac transposition ; Gene tagging ; Germinal excision ; Transgenic Arabidopsis thaliana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Gene tagging in Arabidopsis thaliana using the autonomous Ac (Activator) transposable element has so far been hampered by low frequencies of germinal transposition events. Here we describe a procedure by which the frequency of independent germinal reinsertions has been much improved by a process of long-term selection on kanamycin for the continued growth of tissues in which somatic excisions have occurred. Growth on artificial media increased the somatic excision frequency, and the long-term selection procedure channelled somatic transposition events into the germline. This resulted in an overall germinal excision frequency in the progeny of longterm selected plants of 15%, as confirmed by Southern blotting, with 63% of the plants bearing excision events having detectable reinsertions of the Ac element. This compares with a germinal excision frequency of approximately 1% when no long-term selection is employed. However, offspring from individual plants tended to have identical germinal Ac reinsertion patterns, thus the critical parameter for evaluating the system for tagging purposes is the frequency of individual plants yielding offspring with reinsertions, which was 64%. This high frequency, when coupled to the enhanced germinal transposition rate overall, easily allows the generation of a large population of plants with independent reinsertions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00211042
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  • 5
    Electronic Resource
    Electronic Resource
    The Arabidopsis PHD-finger protein SHL is required for proper development and fertility (2000)
    Springer
    Molecular genetics and genomics 264 (2000), S. 363-370 
    Details
    ISSN: 1617-4623
    Keywords: PHD finger BAH domain Transcription factor Nuclear localization Arabidopsis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. The SHL gene from Arabidopsis thaliana encodes a small nuclear protein that contains a BAH domain and a PHD finger. Both domains are found in numerous (putative) transcriptional regulators and chromatin-remodeling factors. Different sets of transgenic lines were established to analyze the physiological relevance of SHL. SHL expression driven by the CaMV 35S promoter results in reduced growth, early flowering, early senescence, and impaired flower and seed formation. Antisense inhibition of SHL expression gives rise to dwarfism and delayed development. In-frame N-terminal fusion of the SHL protein to β-glucuronidase (GUS) directs GUS to the nucleus of stably transformed Arabidopsis plants. Thus, SHL encodes a novel putative regulator of gene expression, which directly or indirectly influences a broad range of developmental processes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004380000313
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  • 6
    Electronic Resource
    Electronic Resource
    Construction and characterization of the IGF Arabidopsis BAC library (1998)
    Springer
    Molecular genetics and genomics 258 (1998), S. 562-570 
    Details
    ISSN: 1617-4623
    Keywords: Key wordsArabidopsis thaliana ; Genomic library ; Bacterial artificial chromosome ; pBeloBAC-Kan ; Repetitive sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A bacterial artificial chromosome (BAC) library has been established for Arabidopsis thaliana (ecotype Col-0) covering about seven haploid nuclear genome equivalents. This library, called the Institut für Genbiologische Forschung (IGF) BAC library, consists of 10 752 recombinant clones carrying inserts (generated by partial EcoRI digestion) of an average size of about 100 kb in a modified BAC vector, pBeloBAC-Kan. Hybridization with organellar DNA and nuclear repetitive DNA elements revealed the presence of 1.1% clones with mitochondrial DNA, 0.2% clones with plastid DNA, 3.2% clones with the 180 bp paracentromeric repeat, 1.6% clones with 5S rDNA, and 10.8% clones with the 18S-25S rDNA repeat. With its extensive genome coverage, its rather uniformly sized inserts (80 kb 〈85% 〈120 kb) and low contamination with organellar DNA, this library provides an excellent resource for A. thaliana genomic mapping, map-based gene cloning, and genome sequencing.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004380050769
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