Regeneration of Transgenic Rose (Rosa hybrida) Plants from Embryogenic Tissue

Friable embryogenic tissues (FET) were recovered from filament cultures of rose, Rosa hybrida cv. Royalty, and cocultured with Agrobacterium tumefaciens or A. rhizogenes harboring a vector containing neomycin phosphotransferase II (NPTII) and -glucuronidase (GUS) or firefly luciferase (LUC) genes. Putative transformed colonies were selected on kanamycin. Fifty−60 transformed embryogenic callus lines were reproducibly obtained from each gram of FET inoculated with Agrobacterium. Transformed embryogenic calli were transferred to maturation media to form somatic embryos, which subsequently produced flowering plants. The transgenic nature of the plants was confirmed by enzyme assays, polymerase chain reaction, and Southern hybridization. Over 100 transgenic plants have been established in soil and flowered in the greenhouse. This procedure facilitates introduction of desirable genes, especially those controlling flower color, into commercial cultivars of rose.

1. Van der Krol, A., Lenting, P.E., Veenstra, J., Van der Meer, I.M., Koes, R.E., Gerats, A.G.M., Mol, J.N.M. and Stuitje, A.R. 1988. An anti-sense chalcone synthase gene in transgenic plants inhibits flower pigmentation. Nature 333: 866−869. | Article | ChemPort |
2. Smith, C.J.S., Watson, C.F., Ray, J., Bird, C.R., Morris, P.C., Schuch, W. and Grierson, D. 1988. Antisense RNA inhibition of polygalacturonase gene expression in transgenic tomatoes. Nature 334: 724−726. | Article | ISI | ChemPort |
3. Broglie, K., Chet, I., Holliday, M., Cressman, R., Biddle, P., Knowlton, S., Mauvais, C.J. and Broglie, R. 1991. Transgenic plants with enhanced resistance to the fungal pathogen Rhizoctonia solani. Science 254: 1194−1197. | ISI | ChemPort |
4. Robinson, K. and Firoozabady, E. 1993. Transformation of floricultural crops. Scientia Horticulturae 55: 83−99. | Article | ISI | ChemPort |
5. Broertjes, C. and van Harten, A.M. 1978. Roses, p. 178−185. In: Developments in Crop Science, (2). Elsevier, Amsterdam.
6. Noriega, C. and Söndahl, M. 1991 Somatic embryogenesis in hybrid tea roses. Bio/Technology 9: 991−993. | ISI |
7. de Wit, J.C., Esendam, H.F., Honkanen, J.J. and Tuominen, U. 1990. Somatic embryogenesis and regeneration of flowering plants in rose. Plant Cell Reports 9: 456−458. | ChemPort |
8. Burger, D.W., Liu, L., Zary, K.W. and Lee, C.I. 1990. Organogenesis and plant regeneration from immature embryo of Rosa hybrida L. Plant Cell, Tissue and Organ Culture 21: 147−152. | ISI |
9. Ishioka, N. and Tanimoto, S. 1990. Plant regeneration from Bulgarian rose callus. Plant, Cell, Tissue and Organ Culture 22: 197−199. | ISI |
10. Lloyd, D., Roberts, A.V. and Short, K.C. 1988. The induction in vitro of adventitious shoots in Rosa. Euphytica 37: 31−36. | ISI |
11. Hoekema, A., Hirsch, P.R., Hooykaas, P.J.J. and Shilperoort, R.A. 1983. A binary plant vector strategy based on separation of vir- and T-region of the Agrobaaerium tumefaciens Ti-plasmid. Nature 303: 179−180. | ISI | ChemPort |
12. Birot, A-M., Bouchez, D., Casse-Delbart, F., Durand-Tardif, M., Jouanin, L., Pautot, V., Robaglia, C., Tepfer, D., Tepfer, M., Tourneur, J. and Vilaine, F. 1987. Studies and uses of the Ri plasmids of Agrobacterium rhizogenes. Plant Physiol. Biochem. 25: 323−335. | ISI | ChemPort |
13. Lemieux, C.S., Firoozabady, E. and Robinson, K.E.P.R. 1990. Agrobacterium-mediated transformation of chrysanthemum, p. 150−155. Proc. Eucarpia Symp. on Integration of in vitro techniques in ornamental plant breeding. Jong, J. de (Ed.). (CPO Ctr. Pl. Breed. Res., Wageningen, The Netherlands).
14. Hood, E.E., Jen, G., Kayes, L., Kramer, J., Fraley, R.T. and Chilton, M.-D. 1984. Restriction endonucleasc map of pTiBo542, a potential Ti plasmid vector for genetic engineering of plants. Bio/Technology 2: 702−709. | ISI | ChemPort |
15. Jefferson, R.A., Kavanagh, T.A. and Bevan, M.W. 1987. GUS fusions: -glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6: 3901−3907. | PubMed  | ISI | ChemPort |
16. Lu, C.-Y., Nugent, G., Wardley-Richardson, T., Chandler, S.F., Young, R. and Dalling, M.J. 1991. Agrobacterium-mediated transformation of carnation (Dianthus caryophyllus L.). Bio/Technology 9: 864−868. | ISI | ChemPort |
17. McGranahan, G.H., Leslie, C.A., Uratsu, S.L., Martin, L.A. and Dandekar, A.M. 1988. Agrobacterium-mediated transformation of walnut somatic embryos and regeneration of transgenic plants. Bio/Technology 6: 800−804. | ISI | ChemPort |
18. Mathews, H., Litz, R.E., Wilde, D.E., Merkle, S.A., Wetzstein, H.Y. 1992. Stabe integration and expression of -glucuronidase and NPT II genes in mango somatic embryos. In Vitro 28P: 172−178. | ISI | ChemPort |
19. D'Halluin, K., Bossut, M., Bonne, E., Mazur, B., Leemans, J. and Botterman, J. 1992. Transformation of sugarbeet (Beta vulgaris L.) and evaluation of herbicide resistance in transgenic plants. Bio/Technology 10: 309−314. | ChemPort |
20. Firoozabady, E. and Galbraith, D.W. 1984. The effects of cell wall and cell cycle on crown gall tumorigenesis. HortScience 18: 618. | ISI |
21. Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A. and Struhl, K. 1992. Current Protocols in Molecular Biology. John Wiley & Sons, Media PA.
22. Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15: 473−497. | ISI | ChemPort |
23. Chu, C.C., Wang, J.J. and Sun, J.S. 1975. Establishment of an efficient medium for anther culture of rice through comparative experiments of the nitrogen sources. Scientia Sinica 18: 659−668. | ISI |
24. Ow, D.W., Wood, K.V., DeLuca, M., de Wet, J.R., Helsinki, D.R. and Howell, S.H. 1986. Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Science 234: 856−859. | ISI | ChemPort |
25. Ditta, G., Stanfeld, S., Corbin, D. and Helenski, D.R. 1980. Broad host range DNA cloning system for gram-negative bacteria: Construction of a gene bank of Rhizobium meliloti. Proc. Nat. Acad. Sci. USA 77: 7347−7351. | PubMed  | ChemPort |

	Top

	Top