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  • 1
    Electronic Resource
    Electronic Resource
    Glyphosate selection of gene amplification in suspension cultures of 3 plant species (2001)
    Copenhagen : Munksgaard International Publishers
    Physiologia plantarum 112 (2001), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Stepwise selection was carried out with increasing glyphosate concentrations to produce suspension cultures of Medicago sativa L. (alfalfa), Glycine max L. (Merr.) (soybean) and Nicotiana tabacum L. (tobacco) (two lines) that were at least 100-fold more resistant than the original culture as measured by the I50. The selection process required from 8 to 11 transfers to fresh medium over a total period from 161 to 312 days. The alfalfa and soybean lines contained 62- and 21-fold higher activity levels of the glyphosate target enzyme, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), respectively. The tobacco lines had EPSPS enzyme activity levels more than 800-times higher than the original cultures. The EPSPS gene copy number and mRNA were increased in all of the lines as measured by southern and northern hybridization, respectively. Thus, as has been found before with most glyphosate-resistant suspension cultures, the resistance is caused by high EPSPS enzyme activity due to EPSPS gene amplification. Alfalfa and soybean EPSPS gene amplification and the very high EPSPS enzyme activity increases found in the tobacco cultures have not been reported before. These studies show that EPSPS gene amplification can occur in many plant species to confer glyphosate tolerance.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1034/j.1399-3054.2001.1120411.x
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  • 2
    Electronic Resource
    Electronic Resource
    The use of amino acid analog resistance and plant regeneration ability to select somatic hybrids between Nicotiana tabacum and N. glutinosa (1983)
    Springer
    Molecular genetics and genomics 192 (1983), S. 235-240 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protoplasts isolated from a 5-methyltryptophan resistant nonregenerable Nicotiana tabacum (L.) Xanthi suspension culture were fused via the dextran method (Kameya et al. 1981) with protoplasts isolated from N. glutinosa L. leaf mesophyll cells. Prospective somatic hybrids were selected by their ability to produce chlorophyll and to grow on medium containing 5-methyltryptophan and eventually to regenerate into complete plants. Twenty-eight plants regenerated from selected colonies were classified into three groups: (1) Four plants had intermediate morphology, were sterile, had hybrid isozyme patterns for four different enzymes, and contained fraction 1 protein large subunit patterns indicative of a N. glutinosa chloroplast origin. (2) One plant had N. tabacum morphology, was fertile, had two hybrid isozyme patterns, and contained fraction 1 protein large subunits indicative of the N. tabacum chloroplast type. Although this hybrid contained both parental small subunit patterns of fraction 1 protein, two-dimensional gel electrophoresis showed the N. glutinosa-type small subunit to consist of several molecular weights suggestive of errors in the processing steps. Progeny of the Type 2 hybrid (selfed) contained no N. glutinosa characteristics. (3) Twenty-three plants were N. glutinosa escapes. Chromosome numbers ranged from 60 (some chromosome loss) in two hybrids to 34–35 (extensive chromosome loss) in three hybrids. These results suggest that amino acid analog resistance and regeneration capability can be utilized as complementing markers for selecting and identifying somatic hybrids after protoplast fusion. Mesophyll protoplasts from N. glutinosa can also be regenerated into morphologically normal fertile plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00327672
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  • 3
    Electronic Resource
    Electronic Resource
    Characterization of sand as a support for immobilized enzymes (1976)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 18 (1976), S. 527-543 
    Details
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The potential of sand as a support for immobilized enzymes was investigated by preparing alkylamine sand and devising methods to measure the total number of amine groups present and the fraction available for immobilization of enzymes. Alcohol dehydrogenase (alcohol:NAD oxidoreductase, EC 1.1.1.1) and lactate dehydrogenase (L-lactate: NAD oxidoreductase, EC 1.1.1.27) were immobilized on alkylamine sand, and the stability of the immobilized protein and dehydrogenase activity was measured. Urease (urea amidohyrdrolase, EC 3.5.1.5) was also immobilized on sand to test the applicability of these methods to larger scale immobilizations. Results suggest that sand shows promise as a support for immobilized enzymes.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260180407
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