Abstract
Fertile transgenicTriticale ( ×Triticosecale Wittmack) plants expressing theβ-glucuronidase (uidA) and phosphinothricin acetyltransferase (bar) genes were obtained after microprojectile bombardment of scutellar tissue with the plasmid pDB1 containing theuidA gene under the control of the actin-1 promoter (Act1) from rice and the selectable marker genebar under the control of the CaMV 35S promoter. From 465 bombarded scutella about 4000 plantlets were regenerated; 300 plants survived the selection. These regenerants were screened for enzyme activity by the histological GUS assay and by spraying the plants with a herbicide (Basta). Twenty-five regenerants showed GUS activity and survived repeated Basta spraying. Southern blot analysis showed the presence of both marker genes introduced into the genome of analysed plants.
All transgenic plants were fertile. They were grown to maturity and set seed. Pollen and progeny analyses provided evidence for inheritance of the introduced genes to the next generation.
Similar content being viewed by others
References
Barcelo P, Hagel Ch, Becker D, Martin A, Lörz H: Transgenic cereal (Tritordeum) plants obtained at high efficiency by microprojectile bombardment of inflorescence tissue. Plant J 5: 583–592 (1994).
Barton KA, Brill WJ: Prospects in plant genetic engineering. Science 219: 671–675 (1983).
Becker D, Brettschneider R, Lörz H: Fertile transgenic wheat from microprojectile bombardment of scutellar tissue. Plant J 5: 299–307 (1994).
Birch RG, Franks T: Development and optimalisation of microprojectile systems for plant genetic transformation. Aust J Plant Physiol 18: 453–469 (1991).
Budar F, Thia-Toong L, Van Montagu M, Hernalsteens JP:Agrobacterium-mediated gene transfer results mainly in transgenic plants transmitting T-DNA as a single Mendelian factor. Genetics 114: 303–313 (1986).
Cao J, Duan X, McElroy D, Wu R: Regeneration of herbicide resistant transgenic rice plants following microprojectile mediated transformation of suspension culture cells. Plant Cell Rep 11: 586–591 (1992).
Christou P, Ford TL, Kofron M: Production of transgenic rice (Oryza sativa L.) plants following microprojectile mediated transformation of suspension culture cells. Plant Cell Rep 11: 586–591 (1991).
Czernilowsky AP, Hein R, Becker B, Wirtz U: Studies of the structure and functional organization of foreign DNA integrated into the genome ofNicotiana tabacum. DNA 5: 476–482 (1986).
Davey MR, Rech EL, Mulligan BJ: Direct DNA transfer to plant cells. Plant Mol Biol 13: 273–285 (1989).
Dellaporta SL, Woodand J, Hicks JB: A plant DNA minipreparation: Version II. Plant Mol Biol Rep 4: 19–21 (1983).
Gordon-Kamm WJ, Spencer TM, Mangano ML, Adams TR, Daines RJ, Start WG, O'Brian JV, Chambers SA, Adams JWR, Willetts NG, Rice TB, Mackey CJ, Krueger W, Kausch AP, Lemaux PG: Transformation of maize cells and regeneration of fertile transgenic plants. Plant Cell 2: 603–618 (1990).
D'Halluin K, Bonne E, Bossut M, De Beuckeleer M, Leemans J: Transgenic maize plants by tissue electroporation. Plant Cell 4: 1495–1505 (1992).
Hobbs SLA, Warkentin TD, DeLong CMO: Transgene copy number can be positively or negatively associated with transgene expression. Plant Mol Biol 21: 17–26 (1993).
Hooykaas-Van Slogteren GMS, Hooykaas PJJ, Schilperoort RA: Expression of Ti plasmid genes in monocotyledonous plants infected withAgrobacterium tumefaciens. Nature 311: 763–764 (1984).
Jähne A, Becker D, Lörz H: Regeneration of transgenic, microspore derived, fertile barley. Theor Appl Genet 89: 525–533 (1994).
Li L, Rongda Q, de Kochko A, Faquet C, Beachy RN: An improved rice transformation system using the biolostic method. Plant Cell Rep 12: 250–255 (1993).
Lukaszewski AJ, Curtis Ch: Transfer of theGlu-D1 gene from chromosome 1D of breadwheat to chromosome 1A in hexaploidTriticale. Plant Breed 112: 177–182 (1994).
McCabe DE, Swain WF, Martinell BJ, Christou P: Stable transformation of soybean (Glycine max) by particle acceleration. Bio/technology 6: 923–926 (1988).
McElroy D, Zhang W, Cao J, Wu R: Isolation of an efficient actin promoter for use in rice transformation. Plant Cell 2: 163–171 (1990).
Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).
Nehra NS, Chibbar RN, Leung N, Caswell K, Mallard C, Steinhauer L, Boga M, Kartha KK: Self-fertile transgenic wheat plants regenerated from isolated scutellar tissues following microprojectile bombardment with two distinct gene constructs. Plant J 5: 285–297 (1994).
Neuhaus-Url G, Neuhaus G: The use of the nonradioactive digoxigenin chemiluminescent technology for plant genomic Southern blot hybridisation: a comparison with radioactivity. Transgen Res 2: 115–120 (1993).
Pauk J, Manninen O, Mattila I, Salo Y, Pulli S: Androgenesis in hexaploid spring wheat F2 populations and their parents using a multiple-step regenerationsystem. Plant Breed 107: 18–27 (1991).
Potrykus I: Gene transfer to plants: assessment and perspectives. Physiol Plant 79: 125–134 (1990).
Rathore KS, Chowdhury VK, Hodges TK: Use ofbar as a selectable marker gene and for the production of herbicide-resistent rice from protoplasts. Plant Mol Biol 21: 871–884 (1993).
Sanford JC, Klein TM, Wolf ED, Allen N: Delivery of substances into cells and tissues using a particle bombardment process. J Part Sci Technol 5: 27–37 (1987).
Somers DA, Rines HW, Gu W, Kaeppler HF, Bushnell WR: Fertile transgenic oat plants. Bio/technology 10: 1589–1594 (1992).
Spencer TM, O'Brien JV, Start WG, Adams TR: Segregation of transgenes in maize. Plant Mol Biol 18: 201–210 (1992).
Stolarz A, Lörz H: Somatic embryogenesisin vitro manipulation and plant regeneration from immature embryos of hexaploidTriticale ( ×Triticosecale Wittmack). Z Pflanzenzüchtung 96: 353–362 (1986).
Stolarz A: Cell and protoplast culture Somatic embryogenesis and transformation studies in different formes of ×Triticosecale Wittmack. In: Proceedings of the Second InternationalTriticale Symposium, Passo Fundo, Brazil, pp. 286–289 (1990).
Tomes D, Weissinger A, Ross M, Higgins R, Drummond B, Schaaf S, Malone-Schoneberg J, Stabell M, Flynn P, Anderson J, Hovard J: Transgenic tobacco plants and their progeny derived by microprojectile bombardment of tobacco leaves. Plant Mol Biol 14: 261–268 (1990).
Vasil V, Castillo AM, Fromm ME, Vasil IK: Herbicide resistant fertile transgenic wheat plants obtained by microprojectile bombardment of regenerable embryogenic callus. Bio/technology 10: 667–674 (1992).
Walters DW, Vetsch CS, Potts DE, Lundquist RC: Transformation and inheritance of hygromycin phosphotransferase gene in maize plants. Plant Mol Biol 18: 189–200 (1992).
Wan Y, Lemaux PG: Generation of large numbers of independently transformed fertile barley plants. Plant Physiol 104: 37–48 (1994).
Weeks JT, Anderson OD, Blecchl AE: Rapid production of multiple independant lines of fertile transgenic wheat (Triticum aestivum). Plant Physiol 102: 1077–1084 (1993).
Zimny J, Lörz H: High frequency of somatic embryogenesis and plant regeneration of rye (Secale cereale L.). Z Pflanzenzüchtung 102: 89–100 (1989).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Zimny, J., Becker, D., Brettschneider, R. et al. Fertile, transgenicTriticale ( ×Triticosecale Wittmack). Mol Breeding 1, 155–164 (1995). https://doi.org/10.1007/BF01249700
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01249700