Clones from a shooty tobacco crown gall tumor II: irregular T-DNA structures and organization, T-DNA methylation and conditional expression of ... View Full Text


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Article Info

DATE

1986-07

AUTHORS

R. Peerbolte, K. Leenhouts, G. M. S. Hooykaas-van Slogteren, G. J. Wullems, R. A. Schilperoort

ABSTRACT

Transformed clones from a shooty tobacco crown gall tumor, induced byAgrobacterium tumefaciens strain LBA1501, having the auxin locus of the TL-region inactivated by a Tn1831 insertion, were investigated for their T-DNA structure and expression. It has been described previously (28) that in addition to clones with an expected phenotype (phytohormone independent growth in tissue culture (Aut+), shoot regeneration (Reg+) and octopine synthesis (Ocs+)), clones were obtained with an aberrant phenotype. One of these clones, TSO38, is Aut+Reg+ but shows little or no octopine synthesis activity (Ocs-). Subclones of TSO38, however, are either Ocs- or Ocs+. Ocs- shoots become Ocs+ under certain states of differentiation, indicating that the octopine synthase gene is present. The fact that in the Ocs- subclones the octopine synthase gene is not expressed, is probably due to DNA methylation (29). The present paper describes that shoots derived from both an Ocs+ and an Ocs- subclone of TSO38, which were negative for the presence of mannopine (Mas-) and agropine (Ags-), became Mas+Ags+ after culturing on medium containing the hypomethylating agent 5-azacytidine. This means that both in the Ocs- line and in the Ocs+ line expression of TR-DNA opine genes most likely was hampered by DNA methylation. The T-DNA structures of an Ocs- and an Ocs+ TSO38 subclone proved to be identical and surprisingly complex. No intact copy of Tn1831 was present. TL-DNA and TR-DNA segments, present in high copy numbers, were truncated; several T-DNA segments existed in tandem arrangements. When DNA from an Ocs+ and an Ocs- subclone of TSO38 were compared for cleavability by the methylation sensitive restriction enzymes HpaII and MspII, differences were detected, but it became also clear that both lines contained methylated T-DNA segments. This indicates that the Ocs- and the Ocs+ TSO38 subclones differ only quantitatively in respect to degree of T-DNA methylation. More... »

PAGES

285-299

References to SciGraph publications

  • 1983-06. Nopaline T-DNA is maintained during regeneration and generative propagation of transformed tobacco plants in MOLECULAR GENETICS AND GENOMICS
  • 1984-11. Silent T-DNA genes in plant lines transformed by Agrobacterium tumefaciens are activated by grafting and by 5-azacytidine treatment in PLANT MOLECULAR BIOLOGY
  • 1981-08. Sequence specificity of methylation in higher plant DNA in NATURE
  • 1982-02. Agrobacterium rhizogenes inserts T-DNA into the genomes of the host plant root cells in NATURE
  • 1985-07. Transformation of plant protoplasts with DNA: cotransformation of non-selected calf thymus carrier DNA and meiotic segregation of transforming DNA sequences in PLANT MOLECULAR BIOLOGY
  • 1986-07. Clones from a shooty tobacco crown gall tumor I: deletions, rearrangements and amplifications resulting in irregular T-DNA structures and organizations in PLANT MOLECULAR BIOLOGY
  • 1973-07. Streptomycin-resistant Plants from Callus Culture of Haploid Tobacco in NATURE
  • 1983-11. Nucleotide sequence of the T-DNA region from theA grobacterium tumefaciens octopine Ti plasmid pTi15955 in PLANT MOLECULAR BIOLOGY
  • 1983-11. Clonal analysis of heterogeneous crown gall tumor tissues induced by wild-type and shooter mutant strains ofAgrobacterium tumefaciens-expression of T-DNA genes in PLANT MOLECULAR BIOLOGY
  • 1984-11. Changes in T-DNA methylation and expression are associated with phenotypic variation and plant regeneration in a crown gall tumor line in MOLECULAR GENETICS AND GENOMICS
  • 1984-06. Agrobacterium tumefaciens T-DNA in the yeast Saccharomyces cerevisiae in MOLECULAR GENETICS AND GENOMICS
  • 1980-09. The expression of tumour markers in intraspecific somatic hybrids of normal and crown gall cells from Nicotiana tabacum in THEORETICAL AND APPLIED GENETICS
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    http://dx.doi.org/10.1007/bf00752901

    DIMENSIONS

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    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/24302370


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