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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 68 (1986), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The in vivo metabolism of [3H]-tryptophan (TRP), [14C]-indole-3-acetaldoxime (IAOX), and [14C]-indole-3-acetic acid (IAA) has been studied in non-transformed and Agrobacterium tumefaciens T 37 transformed cells of Nicotiana tabacum L. cv. White Burley. Metabolites were analyzed by thin-layer chromatography (TLC) and high pressure liquid chromatography (HPLC). TRP was converted to IAOX and indole-3-ethanol (IEOH) in both cell types, while indole-3-acetamide (IAAM) occurred exclusively in transformed cells. The formation of IEOH was higher in transformed cells. IAOX was metabolized to IEOH and IAA in both tissues, whereas only in transformed cells some conversion to IAAM occurred. Furthermore, the conversion of the aldoxime to IAA was higher in transformed cells. Labelled IAA became rapidly conjugated to indole-3-acetylaspartic acid (IAAsp) in both cell types, the transformed cells showing a 100% increased uptake of IAA. In vitro incubations of [14C]-IAOX with desalted extracts from transformed cells showed no significant enzymatic conversion to IAAM, whereas in the presence of flavin adenine dinucleotide (FAD) some chemical conversion was observed. The present results suggest that in an established line of transformed cells the integration of the T-DNA leads to a permanent transcription of its own auxin genes. Furthermore, the in vivo studies suggest that the host cell's capacity for IAA biosynthesis and conjugation may be altered quantitatively. The potential role of IAOX in both transformed and non-transformed cells is discussed.
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  • 2
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The transition from resting to mitotically active potato tissue (excised disks) is accompanied by and virtually dependent on transcription, translation and increased metabolic activity. Thus enhanced RNA- and protein-syntheses are well known features of potato tuber slices, allowing the cells to raise the level of several metabolic pathways (i.e., starch breakdown, pentose phosphate shunt, glycolysis, respiration and others) and ultimately leading to mitotic activity. Mitosis can be totally, yet reversibly suppressed by incubation of the tissue fragments in Tris (hydroxymethyl)-aminomethane buffer. Neither total respiration nor its sensitivity to several inhibitors is affected during blockage of mitosis. Fluctuations in the levels of glucose-6-phosphate, fructose-6-phosphatc, phospho-enolpyruvate and pyruvate are exactly the same in Tris-inhibited as in non-inhibited cells. Thus the Tris ion is not modifying glucose catabolism. The same is true for the time-course of activity of hexokinase, glucose-6-phosphate- and 6-phosphogluconate dehydrogenase, glucosephosphate isomerase, triosephosphate isomerase, enolase, phosphoglyceromutase and “malic enzyme”. Although the rapid decrease in activity of phosphoglucomutase, aldolase, pyruvate kinase and glutamic-pyruvic transaminase after slicing (characteristic for non-inhibited cells) does not occur in Tris-treated tissue, these small differences are not thought to be causative in inhibition of mitosis.The results indicate, that nucleic acid metabolism is more likely the target of the Tris-ion.
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  • 3
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Dormant tuber tissue of Jerusalem artichoke (Helianthus tuberosus L.) can be stimulated by wounding to initiate RNA and protein synthesis. No DNA synthesis or cell divisions occur unless an auxin is provided. Changes in polysomal profiles and levels of Poly(A)+-RNA in response to wounding and auxin treatment were studied. Polysomes were isolated at various times after excision and incubation of tissue in the presence or absence of 10−5M 2,4-dichlorophenoxyacetic acid. Polysomal profiles were studied by sucrose density gradient centrifugation. Dormant tissue contained ribosomes mainly in monosome form. Within 4 h of excision, a significant increase in the polysomal fraction was observed both in control and auxin-treated tissue. Increases in polysomes continued during the next 20 h. Poly(A)+-RNA was isolated from total polysomal RNA by oligo(dT)-cellulose column chromatography. There was a large increase in the amount of poly(A)+-RNA within 4 h of excision. During the first 43 h of incubation, levels of total polysomal RNA as well as poly(A)+-RNA in tissue treated with 2,4-dichlorophenoxyacetic acid were significantly higher than those in controls.
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 23 (1970), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: During investigations of their metabolism fragments of plant tissue are most commonly incubated in watery solutions in order to replace the conditions within the intact organ approximately. — This paper reports experiments performed to investigate the influence of a hypotonic medium on several parameters of importance in cell metabolism (protein synthesis, enzymatic activity, concentrations of metabolic intermediates, respiration and cell division). — It could be shown that protein synthesis is severely inhibited in hypotonically exposed potato tuber discs. As a consequence the activities of two dehydrogenases of the pentose phosphate shunt (glucose-6-phosphate- and 6-phosphogluconate-dehydrogenase) are very low as compared with tissue in air. Since the activated pentose shunt evidently contributes to the respiratory reaction after derepression of the tissue, lowered activities of these dehydrogenases must consequently result in lower respiratory activity. This is actually realized in washed tissue. Likewise the mitotic activity is inhibited to a considerable extent. — The destruction of permeability barriers within the cell as a consequence of slicing causes leakage of a variety of metabolites such as glucose, glucose-6-phosphate, fructose-6-phosphate and pyruvate. Thus leaching of metabolic active substances is certainly one of the causal factors in the complex reaction of tissue slices exposed to a hypotonic medium, although such important parameters as changed gas conditions and the various effects of altered hydration of cell cytoplasm must be considered.
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 63 (1985), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The in vivo metabolism of L-tryptophan in wound-activated and Agrobacterium tumefaciens, strain C 58, transformed tissues of white potato tubers (Solanum tuberosum L. cv. Saskia) was investigated. The following metabolites of L-tryptophan were identified in both tissues by co-chromatography with authentic standards in several thinlayer chromotography (TLC) and high pressure liquid chromatographic (HPLC) systems: indole-3-acetic acid (IAA), indole-3-acetaldehyde, indole-3-ethanol, indole-3-acetamide and tryptamine. Labelled indole-3-acetaldoxime was only found in transformed tissue. Crown gall tissue generally incorporated [14C]-L-tryptophan into precursors of IAA at a distinctly higher rate than did wound tissue. Tryptamine and indole-3-ethanol accumulated about ten-fold more label in crown gall cells than in cells from wounded tissue. The incorporation of radioactivity into indole-3-acetamide as determined by 2 consecutive TLC systems followed by HPLC analysis was rather low, though consistently observed in both tissues. An indole-3-acetamide hydrolyzing enzyme, the putative product of gene 2 on the T-DNA, could be extracted from the transformed tissue only. The indole-3-ethanol level was 4.3 nmol (g dry weight)−1 and 41 nmol (g dry weight)−1 for wounded tissue and primary crown gall tissue, respectively, as determined by HPLC with a [14C]-labelled internal standard. The experiments are critically discussed in relation to recent reports on a T-DNA encoded enzyme of IAA biosynthesis in crown gall tumors.
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  • 6
    ISSN: 1432-0983
    Keywords: DNA fingerprinting ; Synthetic oligodeoxynucleotides ; Simple repetitive sequences ; Fungal pathotypes ; Ascochyta rabiei
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ascomycete fungus Ascochyta rabiei, an important pathogen of the grain legume crop chickpea (Cicer arietinum L.) in the Mediterranean region, has not been adequately characterized in molecular terms. We therefore used DNA fingerprinting, with synthetic oligodeoxynucleotides complementary to simple repetitive sequences, to pathotype different isolates of the fungus. Six single-spored A. rabiei isolates were first categorized using a host differential set of nine chickpea genotypes. Seedlings were inoculated under controlled environmental conditions, and disease severity was recorded 9 days after inoculation. DNA was extracted from in vitro-grown mycelia of the six purified fungal isolates, restricted with EcoRI, HinfI, MboII and TaqI, and fingerprinted with radiolabeled (GATA)4, (GTG)5, (CA)8, and (TCC)5, respectively. High levels of polymorphism were detected with optimal enzyme/probe combinations that allow one to discriminate between the isolates. The potential of DNA fingerprinting with simple repetitive sequences can thus be expanded to the identification of fungal races and pathotypes. The characterization of the geographic distribution and genetic variability of pathotypes will facilitate the selection of suitable host cultivars to be grown in specific regions.
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  • 7
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Company
    Nature biotechnology 10 (1992), S. 1030-1035 
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] DNA oligonucleotide and amplification fingerprinting have been successfully used to detect genetic polymorphisms in 15 representative species and cultivars of the genus Musa, comprising AA, AAA, AAAA, AAB, ABB, and BB genotypes. In–gel–hybridization of Hinf I–digested genomic ...
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  • 8
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 327 (1987), S. 529-532 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] Fig. 1 Formation of crown gall tumour on a disk of Dioscorea bulbifera (Yam) bulbil tissue, 90 days after infection with induced Agrobacterium tumefaciens, strain C58 (a) and sterile tumour tissue culture on hormone-free medium 10 months after transfer (6). Tumours were ~1 cm (a) and 3 cm (b) in ...
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  • 9
    ISSN: 1432-0983
    Keywords: Ascochyta rabiei ; DNA fingerprinting ; Simple repetitive sequences ; Genetic diversity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Fifty isolates of Ascochyta rabiei (Pass.) Labr. were hierarchically sampled from four well-separated locations of a single chickpea field in Beja (Tunisia), and single-spored. DNA was isolated from in-vitro-grown mycelia, digested with HinfI or RsaI, and hybridized to a set of synthetic oligonucleotides complementary to simple repetitive sequences. According to the fingerprint patterns derived from the probes (CA)8, (CAA)5, (CAT)5 and (GATA)4, 12 different fungal haplotypes were found at various frequencies within the investigated field. Seven haplotypes were confined to one location only, four occurred at two, one at three, and none at all four locations. Most of the genetic variability originated from diversity within, rather than between, locations. In some cases, more than one haplotype was isolated from the same lesion of a single host plant. Genetic distances between isolates, as calculated from band-sharing data, varied between 0.05 and 0.22. Relatedness between the different haplotypes was evaluated by cluster analysis using UPGMA.
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  • 10
    ISSN: 1432-203X
    Keywords: Achillea ; Simple repetitive sequences ; Oligonucleotide fingerprinting ; RAPD analysis ; Clone identification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two different DNA fingerprinting techniques were applied to a set of Achillea samples (Asteraceae), comprising ten taxa of the medicinally important A. millefolium group and six related species. Field-grown as well as in vitro-micropropagated plants were individually screened for abundance and polymorphism of target sequences recognized by oligonucleotide fingerprinting with 13 different microsatellite-complementary probes. While most probes revealed a high level of intra- and interspecific variability, fingerprints proved to be somatically stable in vegetatively propagated plant material. Analysis of the same samples by polymerase chain reaction with arbitrary 10-mer primers yielded less polymorphic patterns. Because of its higher discriminatory ability, oligonucleotide fingerprinting offers itself as the method of choice for the identification and discrimination of A. asplenifolia and A. roseoalba clones, as well as for monitoring their stability during micropropagation.
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