Abstract
Agrobacterium tumefaciens binary vectors carrying kanamycin resistance gene and either C58 T-DNA gene 4 for cytokinin synthesis or genes 1 and 2 for auxin synthesis were constructed and used for transformation of a short-day tobacco Maryland Mammoth. Kanamycin resistant plants were regenerated from a small fraction of transformed tissue and the presence of T-DNA in their genome was verified by Southern blotting. The level of endogenous cytokinin in plants transgenic for gene 4 and the level of endogenous IAA in those transgenic for genes 1 and 2 increased by more than 100 %. A number of morphological characteristics distinguish them from untransformed controls.
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An, G., Watson, B. D., Stachel, S., Gordon, M. P., Nester, E. W.: New cloning vehicles for transformation of higher plants. - EMBO J.4 : 227–284, 1985.
Barry, G. R., Rogers, S. G., Fraley, R. T., Brand, L.: Identification of a cloned cytokinin biosynthetic gene. - Proc. nat. Acad. Sei USA81: 4776–4782, 1984.
Casse, F., Boucher, G., Julliot, J. S., Michel, N., Denarie, J.: Identification and characterization of large plasmids inRhizobium meliloti and characterization using gel electrophoresis. - J. gen. Microbiol.113 : 229–242, 1979.
Depicker, A., De Wilde, M., De Vos, G., Van Montagu, M., Schell, J.: Molecular cloning of overlapping segments of the nopaline Ti plasmid pTiC58 as a mean to restriction endonuclease mapping. - Plasmid3: 193–211, 1980.
Eder, J., Rovenská, J., Kutáček, M., Čermák, V.: HPLC analysis inAgrobacterium and transformed tobacco cells. - In : Kutáček, M., Bandurski, R. S., Krekule, J. (ed.) : Physiology and Biochemistry of Auxins in Plants. Pp. 389–390. Proc. Int. Symp. Liblice Czechoslovakia 1987. Academia Praha, SPB Acad. Publ., The Hague 1988.
Follin, A., Inzé, D., Genetello, C., Van Montagu, M., Schell, J.: Genetic evidence that the tryptophan- -2-monooxygenase gene ofPseudomonas sevastonii is functionally equivalent to one of the T-DNA genes involved in plant tumor formation byAgrobacterium tumefaciens. - Mol. gen. Genet.201 : 178–185, 1985.
Gelvin, S. B., Karcher, S. J., Di Rita, V. J.: Methylation of the T-DNA inAgrobacterium tumefaciens and in several crown gall tumors. - Nucl. Acids Res.11: 159–174, 1983.
Hoekema, A. Hirsch, P. R., Hooykaas, P. J. J., Schilperoort, R. A.: A binary plant vector strategy based on separation of vir - and T-region of theAgrobacterium tumefaciens Ti plasmid. - Nature303:179–180, 1983.
Holsters, M., De Waele, D., Depicker, A., Van Montagu, M., Schell, J.: Transfection and transformation ofAgrobacterium tumefaciens. - Mol. gen. Genet.163 :181–187, 1978.
Horsch, R. B., Fry, J. E., Hoffmann, N. L., Eichholz, D., Rogers, S. G., Fraley, R. T.: A simple and general method for transferring genes into plants. - Science227:1229–1231, 1985.
Maniatis, T., Fritsch, E. F., Sambrook, J.: Molecular Cloning : A Laboratory Manual. - Cold Spring Harbor Lab., Cold Spring Harbor, New York 1982.
Mazin, V. V., Shashkova, L. S., Andreev, L. I.: [Specificity ofAmaranthus response to kinetin.] In Russ.- Dokl. Akad. Nauk SSSR231: 506–509, 1976.
Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassay with tobacco tissue cultures. - Plant Physiol.15 : 473–497, 1962.
Odell, J. T., Nagy, F., Chua, N. H.: Variability in 35S promoter expression between independent transformants. - In : Key, J. L., Mc Intosh, L. (ed.) : Plant Gene Systems and their Biology. Pp. 321–329. A. R. Liss Inc., New York 1987.
Ondřej, M., Eder, J., Hrouda, M., Machácková, I., Vlasák, J.: Free auxin level and inheritance of introduced markers in tobacco transformed by binary vector based on A4 Ri plasmid. - Biol. Plant.31: 286–291, 1989a.
Ondřej, M., Hrouda, M., Karavajko, N. N., Matoušek, J., Mikulovic, T. P., Pavingerová, D., Vlasák, J.: Transformation byAgrobacterium vectors and the study of functions of plant hormones. - In : Krekule, J., Seidlová F. (ed.) : Signals in Plant Development. Proc. 14th Biochem. Congress, Praha 1988. Pp. 73–89. SPB Academic Publishing, The Hague 1989b.
Roger, S. O., Bendich, A. J.: Extraction of DNA from milligram amounts of fresh, herbacious and mummified plant tissues.- Plant mol. Biol.5 : 69–73, 1985.
Simpson, J., Timko, M. P., Cashmore, A. R., Schell, J., Van Montagu, M., Herrera-Estrella, L.: Light inducible and tissue specific expression of a chimeric gene under control of the 5′-flanking sequence of a pea chlorophylla/b protein binding gene. - EMBO J.4 : 2723–2729, 1985.
Thomashow, L. S., Reeves, S., Thomashow, M. F.: Crown gall oncogenesis : Evidence that a T-DNA gene from theAgrobacterium Ti plasmid pTiA6 encodes an enzyme that catalyzes synthesis of indoleacetic acid. - Proc. nat. Acad. Sei. USA81: 5071–5075, 1984.
Wettstein, D.: Chlorophyll - Letale und der submikroskopische Formwechsel der Piastiden. - Exp. Cell Res.12 : 427–506, 1957.
Willmitzer, L., Dhaese, P., Schreier, P. H., Schmalenbach, W., Van Montagu, M., Schell, J.: Site, localization and polarity of T-DNA encoded transcripts in nopaline crown gall tumors. Common transcripts in octopine and nopaline tumors. - Cell32 : 1045–1056, 1983.
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OndŘej, M., Bavrina, T.V., Dudko, N. et al. Transgenic Tobacco plants with T-DNA Phytohormone synthesis genes. Biol Plant 33, 40–48 (1991). https://doi.org/10.1007/BF02873786
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DOI: https://doi.org/10.1007/BF02873786