Summary
Six octopine tumour lines incited by pTiB6S3, pTiAch5 and pTiA6 on tobacco, Arabidopsis and Petunia were studied by the Southern blotting hybridisation technique in order to define accurately the dimensions of the segments of plasmid origin transferred to the tumourous cell and their organisation in the plant genome. Emphasis has been put on the comparison between octopine and nopaline T-DNAs and on the lines presented here compared with those studied previously (Thomashow et al. 1980).
The lenght of the transferred DNA segment does not depend on the plasmids used, nor on the host plants. The octopine T-DNA organisation in the cell nucleus is significantly different from that of nopaline T-DNAs: tandem arrangements of T-DNA segments could not be detected and the T-DNA itself is much shorter.
The tumour lines described here can be compared to some extent with those studied by another group (Thomashow et al. 1980) by the same technique. However, some differences were observed. The transferred DNA was seen as a unique stretch of about 11 kb present only once per cell. No amplification of any part was noticed in any of these six lines.
Examination of the restriction patterns presented by the boundary fragments of the T-DNA in these lines suggested that some of them were of common origin.
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References
Bomhoff G, Klapwijk PM, Kester HCM, Schilperoort RA, Hernalsteens JP, Schell J (1976) Octopine and nopaline synthesis and breakdown genetically controlled by a plasmid of Agrobacterium tumefaciens. Mol Gen Genet 145:177–181
Chater KJ (1977) A site-specific endodeoxyribonuclease from Streptomyces albus CMI 52766 sharing site specificity with Providencia stuartii endonuclease PstI. Nucl Acids Res 4:1989–1998
Chilton MD, Drummond HJ, Merlo DJ, Sciaky D, Montoya AL, Gordon MP, Nester EW (1977) Stable incorporation of plasmid DNA into higher plant cells: the molecular basis of crown gall tumorigenesis. Cell 11:263–271
Chilton M-D, Drummond MH, Merlo DJ, Sciaky D (1978) Highly conserved DNA of Ti-plasmids overlaps T-DNA maintained in plant tumors. Nature (London) 275:147–149
De Beuckeleer M, De Block M, De Greve H, Depicker A, De Vos R, De Vos G, De Wilde M, Dhaese P, Dobbelaere MR, Engler G, Genetello C, Hernalsteens JP, Holsters M, Jacobs A, Schell J, Seurinck J, Silva B, Van Haute E, Van Montagu M, Van Vliet F, Villarroel R, Zaenen I (1978) The use of the Ti-plasmid as a vector for the introduction of foreign DNA into plants. Proc IVth Int Conf on Plant Pathogenic Bacteria, Angers, I.N.R.A., 115–126
Depicker A, Van Montagu M, Schell J (1978) Homologous DNA sequences in different Ti-plasmids are essential for oncogenicity. Nature (London) 275:150–153
De Vos G, De Beuckeleer M, Van Montagu M, Schell J (1981) Restriction endonuclease mapping of the octopine tumor inducing pTiAch5 of Agrobacterium tumefaciens. Plasmid 6:249–253
Drummond MH, Chilton M-D (1978) Tumor-inducing (Ti) plasmids of Agrobacterium share extensive regions of DNA homology. J Bacteriol 136:1178–1183
Drummond MH, Gordon MP, Nester EW, Chilton M-D (1977) Foreign DNA of bacterial plasmid origin is transcribed in crown gall tumours. Nature (London) 269:535–536
Engler G, Van Montagu M, Zaenen I, Schell J (1977) Homology between Ti-plasmids of Agrobacterium tumefaciens: hybridisation studies using electron microscopy. Biochem Soc Trans 5:930–931
Engler G, Depicker A, Maenhaut R, Villarroel-Mandiola R, Van Montagu M, Schell J (1981) Physical mapping of DNA base sequence homologies between an octopine and a nopaline Ti-plasmid of Agrobacterium tumefaciens. J Mol Biol, in press
Firmin JL, Fenwick GR (1978) Agropine — a major new plasmid-determined metabolite in crown gall tumours. Nature (London) 276:842–844
Gurley WB, Kemp JD, Albert MJ, Sutton DW, Callis J (1979) Transcription of Ti plasmid-derived sequences in three octopine-type crown gall tumor lines. Proc Natl Acad Sci USA 76:2828–2832
Guyon P, Chilton M-D, Petit A, Tempé J (1980) Agropine in “null type” crown gall tumors: evidence for the generality of the opine concept. Proc Natl Acad Sci USA 77:2693–2697
Hamilton RH, Fall MZ (1971) The loss of tumor initiating ability in Agrobacterium tumefaciens by incubation at high temperature. Experientia 27:229–230
Hepburn AG, Hindley J (1979) Regions of DNA sequence homology between an octopine and nopaline Ti plasmid of A. tumefaciens. Mol Gen Genet 169:163–172
Holsters M, Silva B, Van Vliet F, Genetello C, De Block M, Dhaese P, Depicker A, Inzé D, Engler G, Villarroel R, Van Montagu M, Schell J (1980) The functional organization of the nopaline A. tumefaciens plasmid pTiC58. Plasmid 3:212–230
Koekman BP, Ooms G, Klapwijk PM, Schilperoort RA (1979) Genetic map of an octopine Ti-plasmid. Plasmid 2:347–357
Lemmers M, De Beuckeleer M, Holsters M, Zambryski P, Depicker A, Hernalsteens JP, Van Montagu M, Schell J (1980) Internal organization, boundaries and integration of Ti-plasmid DNA in nopaline crown gall tumours. J Mol Biol 144:353–376
Merlo DJ, Nutter RC, Montoya AL, Garfinkel DJ, Drummond MH, Chilton M-D, Gordon MP, Nester EW (1980) The boundaries and copy numbers of Ti plasmid T-DNA vary in crown gall tumors. Mol Gen Genet 177:637–643
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plantarum 15:473–497
Petit A, Delhaye S, Tempé J, Morel G (1970) Recherches sur les guandines des tissus de crown gall. Mise en évidence d'une relation biochimique spécifique entre les souches d'Agrobacterium et les tumeurs qu'elles induisent. Physiol Vég 8:205–213
Rigby PWW, Dieckmann M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251
Sciaky D, Montoya AL, Chilton M-D (1978) Fingerprints of Agrobacterium Ti plasmids. Plasmid 1:238–253
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–518
Thomashow MF, Nutter R, Montoya AL, Gordon MP, Nester EW (1980) Integration and organization of Ti-plasmid sequences in crown gall tumors. Cell 19:729–739
Willmitzer L, De Beuckeleer M, Lemmers M, Van Montagu M, Schell J (1980) DNA from Ti plasmid present in nucleus and absent from plastids of crown gall plant cells. Nature (London) 287:359–361
Willmitzer L, Otten L, Simons G, Schmalenbach W, Schröder J, Schröder G, Van Montagu M, De Vos G, Schell J (1981) Nuclear and polysomal transcripts of T-DNA in octopine crown gall suspension and callus cultures. Mol Gen Genet 182:255–262
Yadav NS, Postle K, Saiki RK, Thomashow MF, Chilton M-D (1980) T-DNA of a crown gall teratoma is covalently joined to host plant DNA. Nature (London) 287:458–461
Yang F, Montoya AL, Merlo DJ, Drummond MH, Chilton M-D, Nester EW, Gordon MP (1980) Foreign DNA sequences in crown gall teratomas and their fate during the loss of the tumorous traits. Mol Gen Genet 177:707–714
Zambryski P, Holsters M, Kruger K, Depicker A, Schell J, Van Montagu M, Goodman HM (1980) Tumor DNA structure in plant cells transformed by A. tumefaciens. Science 209:1385–1391
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Communicated by G. Melchers
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De Beuckeleer, M., Lemmers, M., De Vos, G. et al. Further insight on the transferred-DNA of octopine crown gall. Molec. Gen. Genet. 183, 283–288 (1981). https://doi.org/10.1007/BF00270630
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DOI: https://doi.org/10.1007/BF00270630