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Publications (4)13.42 Total impact

  • Article: Plant mutants with altered responses to cytokinins
    E. Miklashevichs, R. Walden
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    ABSTRACT: Compared with other phytohormones, relatively little is known about the biosynthesis and the molecular action of cytokinins. A number of plant mutants with changed response to, or requirement for cytokinins are now available. It is likely that the biochemical and molecular genetic analysis of these mutants will contribute considerably to our understanding of the nature of cytokinin action.
    Physiologia Plantarum 04/2006; 100(3):528 - 533. · 3.11 Impact Factor
  • Article: T-DNA tagging reveals a novel cDNA triggering cytokinin- and auxin-independent protoplast division.
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    ABSTRACT: Activation T-DNA tagging was used to generate four cytokinin-independent (cyi1-4) tobacco cell lines. Plants regenerated from the mutant lines displayed similar phenotypes: reduced apical dominance, poorly developed roots, delayed growth and flowering, and male and female sterility. Tissue culture experiments demonstrated that the mutations in the different lines uncouple cell proliferation from the effects of both cytokinin and auxin. No significant increase of cytokinin or auxin was found in transgenic calli in comparison with untransformed callus. The functional plant sequence tagged in one of the mutant lines, cyi1, was used to isolate an active cDNA, cyi1a, that was able to trigger cytokinin- and auxin-independent protoplast division. Northern analysis shows that the transcript corresponding to cyi1a accumulates to high levels in the untransformed protoplasts shortly before the onset of cell division, and that these levels decrease when protoplasts reach maximum rates of cell division. A small putative open reading frame, starting with the first ATG in cyi1a and encoding a 22 amino acid peptide, has the same activity in tobacco protoplasts as the whole cDNA. This activity is destroyed by a frame shift mutation. Apparently cyi1a encodes a peptide which participates in the events downstream of a joint point of cytokinin and auxin action leading to cell division.
    The Plant Journal 10/1997; 12(3):489-98. · 6.16 Impact Factor
  • Article: Activation tagging: a means of isolating genes implicated as playing a role in plant growth and development.
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    ABSTRACT: Activation T-DNA tagging has been used to generate a variety of tobacco cell lines selected by their ability to grow either in the absence of auxin or cytokinin in the culture media, or under selective levels of an inhibitor of polyamine biosynthesis. The majority of the cell lines studied in detail contain single T-DNA inserts genetically co-segregating with the selected phenotype. While most of the plants regenerated from the mutant cell lines appear phenotypically normal, several display phenotypes which could be inferred to result from disturbances in the content, or the metabolism, of auxins and cytokinins, or polyamines. The tagging vector is designed to allow the isolation of tagged plant genes by plasmid rescue. Confirmation that the genomic sequence responsible for the selected phenotype has indeed been isolated is provided by PEG-mediated protoplast DNA uptake of rescued plasmids followed by selection for protoplast growth under the original selective conditions. Several plasmids have been rescued from the mutant lines which confer on transfected protoplasts the ability to grow either in the absence of auxin or cytokinin in the culture media, or under selective levels of an inhibitor of polyamine biosynthesis. This review describes the background to activation tagging and our progress in characterizing the genes that have been tagged in the mutant lines we have generated.
    Plant Molecular Biology 01/1995; 26(5):1521-8. · 4.15 Impact Factor
  • Article: Re-evaluation of phytohormone-independent division of tobacco protoplast-derived cells
    Plant Journal 17 (1999) 5.