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  • 1
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Kidney vetch seedlings were induced to form hairy roots by inoculating their mesocotyls with the wild-type strain 15834 of Agrobacterium rhizogenes or with the A. tumefaciens strain C58C1 containing a binary vector system (the pRiA4b as a helper and the vector pCB1346 bearing a pTiC58-derived isopentenyl transferase gene (ipt, cytokinin biosynthetic gene) under control of its native regulatory sequences). Transgenic lines of three distinct phenotypes were selected: (i) Typically, the pRi15834-transformed tissues were stabilized in vitro and maintained for long periods as aseptic, fast-growing, hormone-independent, plagiotropic hairy root cultures which never regenerated shoots and lost the ability to synthesize opines. Their genomic DNA contained both the TL- and the TR-DNA. (ii) One of the HR-lines transgenic for the T-DNA of pRi15834 (named 52AV34) started to regenerate spontaneously into teratomous shoots. The shoots were found to produce opines and both the TL and TR parts of T-DNA were found to be partly deleted and/or rearranged. They contained phytohormones in similar levels as those found in seed-born shoots. (iii) A practically identical morphogenic response as in the line 52AV34 was observed in the clone 27AV46. However, its shooty, dark-green, slow-growing teratomas were proven to be kanamycin-resistant, opine-producing, and double-transformed by the pRiA4b sequences and the ipt gene. They over-produced auxins as well as cytokinins (mainly indoleacetylaspartic acid and ribosides of zeatin and isopentenyladenine).
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5044
    Keywords: Nicotiana tabacum L. ; plant nuclease ; callus tissue ; plant regeneration ; callus enzymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract It was determined using electrophoresis in polyacrylamide gels containing native DNA or RNA that sugar non-specific nuclease active at pH 5.2 was expressed in tobacco callus. The nuclease had a relative molecular mass of about 34.6 kDaltons and degraded substrates in the following order of decreasing rate: denaturated DNA〉poly dA〉UV-irradiated native DNA〉native DNA〉alkylated native DNA〉apurinated native DNA〉poly dG≳poly dC. The nuclease activity changed during callus growth and plant regeneration, but no developmental changes in electrophoretic patterns were detected. The increase in specific DNAse activity of nuclease was maximal in the exponential phase of callus growth on both growth and regeneration media, except for activity in the cytokinin-independent cell strain grown on growth medium. The specific DNAse activity of nuclease decreased during the bud formation period, while total DNAse activity calculated per mg of dry weight was slightly higher in vegetative buds (9.1U) than in undifferentiated tissue of callus (8.5U). Specific DNAse activity was, on the average, several hundred-fold lower in the vegetative tissues of flowering tobacco plants than in calluses in the exponential phase of growth.
    Type of Medium: Electronic Resource
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