Abstract
Kanamycin resistance gene was introduced into tobacco and Atropa belladonna cells by binary vectors, based on Agrobacterium, by means of inoculation of seedlings. The plasmid pGA472, which carries chimaeric kanamycin resistance gene expressed in plants was introduced by transformation into A. tumefaciens Bo542, harbouring pTiBo542 plasmid and A. rhizogenes 8196, carrying pRi8196 plasmids and the resulting two strains were used as binary vectors. Tobacco tumors induced by A. tumefaciens Bo542(pGA472) grew as undifferentiated, kanamycin resistant tissues. Those induced by A. rhizogenes 8196(pGA472) differentiated into transformed plants. When cultivated in vitro on 200 μg ml-1 kanamycin medium, they showed yellow green sectoring, which was not selected out during vegetative propagation. Atropa belladonna tissues transformed by both A. tumefaciens Bo542(pGA472) and A. rhizogenes 8196(pGA472) differentiated plants which grew well on 200 μg ml-1 kanamycin as green, non-sectoring plants; sensitive cells obviously did not divide at all. Selection of Atropa belladonna transformed tissues on kanamycin medium is much more efficient than selection of transformed tobacco tissues with introduced kanamycin resistance gene.
Similar content being viewed by others
References
An, G., Watson, B. D., Stachel, S., Gordon, M. P., Nesteb, E. W.: New cloning vehicles for transformation of higher plants. - EMBO J.4: 277–284, 1985.
Casse, E., Bouchkeb, C., Julliot, J. B., Michel, M., Denaeie, J.: Identification and characterization of large plasmids in Rhizobium meliloti using agarose gel electrophoresis.- J. gen. Microbiol.113: 229–242, 1979.
De Block, M., Herrera-Estbella, L., Schell, J., Zambryski, P.: Expression of foreign genes in regenerated plants and their progeny. - EMBO J.3: 1681–1689, 1984.
De Fbamond, A. J., Babton, K. A., Chilton, M. D.: MiniTi: a new vector strategy for plant genetic engineering. - Biotechnology1: 262–269, 1983.
Fraley, R. T., Rogers, S. G., Hobsch, R. B., Sandebs, P. R., Flick, J. S., Adams, P. S., Bittneb, M. L., Bband, L. A., Fink, C. L., Woo, S. C.: Expression of bacterial genes in plant cells. - Proc. nat. Acad. Sci. USA80: 4803–4807, 1983.
Herreea-Estrella, L., Depickeb, A., Van Montagu, M., Scheix, J.: Expression of chimaeric genes transformed into plant cells using a Ti plasmid vector. - Nature303: 209–213, 1983a.
Herrera-Estrella, L., De Block, M., Messens, E., Hernalsteens, J. P., Van Montagu, M., Schell, J.: Chimerio genes as dominant selectable markers in plant cells. - EMBO J.2: 987–995, 1983b.
Hokkema, A., Hirsch, P. R., Hooykaas, P. J. J., Schiukroort, R. A.: A binary plant vector strategy based on separation of vir-and T-region of the Agrobacterium tumefaciens Ti plasmid. — Nature303: 179–180, 1983.
Holstebs, M., De Waele, P., Depickbb, A., Messens, E., Van Montagu, M., Scheix, J.: Transfection and transformation of Agrobacterium tumefaciens. — Mol. gen. Genet.163: 181 to 187, 1978.
Koziel, M. G., Adams, T. L., Hazlet, M. A., Adam, D., Miller, J., Dahlbeck, D., Jayne, S., Staskawicz, B. J.: A cauliflower mosaic virus promoter directs expression of kanamycin resistance in morphogenic transformed plant cells. — J. mol. appl. Genet.3: 549–562, 1984.
Langley, R. A., Kado, C. I.: Studies on Agrobacterium tumefaciens: Conditions for mutagenesis by N-methyl-N’-nitro-N-nitrosoquanidine and relationship of A. tumefaciens mutants to crown gall induction. — Mutation Res.14: 277–286, 1972.
Murashige, T., Skoog, F.: A. revised medium for rapid growth and bioaasays with tobacco tissue cultures. - Physiol. Plant,15: 473–479, 1962.
Ondřej, M., Bísková, R., Vlasák, J.: Binary plant vector based on Ri plasmid and part of T-DNA of the Ti plasmid. - Biol. Plant.28: 265–269, 1986.
Ondřej, M.,Protivá, J.: In vitro culture of crown gall and hairy root tumors of Atropa belladonna: Differentiation and alkaloid production. — Biol. Plant.29, in press, 1987.
Petit, A., David, C., Dahl, G. A., Ellis, J. G., Guyon, P., Casse-Delbart, F., Tempe, J.: Further extension of the opine concept; plasmids in Agrobacterium rhizogenes cooperate for opine degradation. - Mol. gen. Genet.190: 204–211, 1983.
Reiss, R., Sprenoel, R., Will, H., Schaller, H.: A new sensitive method for qualitative and quantitative analysis of neomycin phosphotransferase in crude cell extracts. — Gene30: 217–223, 1984.
Wang, K., Herreba-Estrella, L., Van Montagu, M., Zambkyski, P.: Right 25 bp terminus sequence of the nopaline T-DNA is essential for and determines the direction of T-DNA transfer from Agrobacterium to plant cell genome. — Cell38: 455–462, 1984.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ondrej, M., Vlasák, J. Expression of Kanamycin resistance introduced byAgrobacteriutn binary vector intoNicotiana tabacum andAtropa belladonna . Biol Plant 29, 161–166 (1987). https://doi.org/10.1007/BF02876820
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02876820