Abstract
Zea mays transformants produced by particle bombardment of embryogenic suspension culture cells of the genotype A188 × B73 and selected on kanamycin or bialaphos were characterized with respect to transgene integration, expression, and inheritance. Selection on bialaphos, mediated by thebar orpat genes, was more efficient than selection on kanamycin, mediated by thenptII gene. Most transformants contained multicopy, single locus, transgene insertion events. A transgene expression cassette was more likely to be rearranged if expression of that gene was not selected for during callus growth. Not all plants regenerated from calli representing single transformation events expressed the transgenes, and a non-selectable gene (uidA) was expressed in fewer plants than was the selectable transgene. Mendelian inheritance of transgenes consistent with transgene insertion at a single locus was observed for approximately two thirds of the transformants assessed. Transgene expression was typically, but not always, predictable in progeny plants-transgene silencing, as well as poor transgene transmission to progeny, was observed in some plant lines in which the parent plants had expressed the transgene.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Armstrong CL, Green CE: Establishment and maintenance of friable, embryogenic maize callus and the involvement of L-proline. Planta 164: 207–214 (1985).
Bellini C, Guerche P, Spielmann A, Goujard J, Lesaint C, Caboche M: Genetic analysis of transgenic tobacco plants obtained by liposome-mediated transformation: absence of evidence for the mutagenic effect of inserted sequences in sixty characterized transformants. J Hered 80: 361–367 (1989).
Bennetzen JL: Covalent DNA modifications and the regulation ofMutator element transposition in maize. Mol Gen Genet 208: 45–51 (1987).
Chomet PS, Wessler S, Dellaporta SL: Inactivation of the maize transposable elementActivator (Ac) is associated with its DNA modification. EMBO J 6: 295–302 (1987).
Chu CC, Wang CC, Sun CS, Hsu C, Yin KC, Chy CY, Bi FY: Establishment of an efficient medium for anther culture of rice through comparative experiments on the nitrogen source. Sci Sin (Peking) 18: 659–668 (1975).
Deroles SC, Gardner RC: Expression and inheritance of kanamycin resistance in a large number of transgenic petunias generated byAgrobacterium-mediated transformation. Plant Mol Biol 11: 355–364 (1988).
Deroles SC and Gardner RC: Analysis of the T-DNA structure in a large number of transgenic petunias generated byAgrobacterium-mediated transformation. Plant Mol Biol 11: 365–377 (1988).
Coe EH, Neuffer MG, Hoisington DA: The genetics of corn. In: Sprague GF, Dudley JW (eds) Corn and Corn Improvement, 3rd ed, pp. 83–258. American Society of Agronomy Press, Madison (1988).
D'Halluin K, Bonne E, Bossut M, DeBruekeleer M, Leemans J: Transgenic maize plants by tissue electroporation. Plant Cell 4: 1495–1505 (1992).
Edwards K, Johnstone C, Thompson C: A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucl Acids Res 19: 1349 (1991).
Federoff N, Banks JA, Masson P: Molecular genetic analysis of the maizeSuppressor-mutator element's epigenetic developmental regulatory mechanism. Genome 31: 973–979 (1989).
Feinberg AP, Vogelstein B: A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Analyt Biochem 132: 6–13 (1983).
Fromm ME, Morrish F, Armstrong C, Williams R, Thomas J, Klein TM: Inheritance and expression of chimeric genes in the progeny of transgenic maize plants. Bio/Technology 8: 833–839 (1990).
Gordon-Kamm WJ, Spencer TM, Mangano ML, Adams TR, Daines RJ, Start WG, O'Brien JV, Chambers SA, Adams WR, Willets NG, Rice TB, Mackey CJ, Krueger RW, Kausch AP, Lemaux PG: Transformation of maize cells and regeneration of fertile transgenic plants. Plant Cell 2: 603–618 (1990).
Hamilton DA, Bashe DM, Stinson JR, Mascarhenas JP: Characterization of a pollen-specific genomic clone from maize. Sex Plant Reprod 2: 208–212 (1989).
Hamilton DA, Roy M, Ruenda J, Sindhu RK, Sanford J, Mascarhenas JP: Dissection of a pollen-specific promoter from maize by transient transformation assays. Plant Mol Biol 18: 211–218 (1992).
Hauptmann RM, Vasil V, Ozias-Aikins P, Tabaeizadeh Z, Rogers SG, Fraley RT, Horsch RB, Vasil IK: Evaluation of selectable markers for obtaining stable transformants in the Gramineae. Plant Physiol 86: 602–608 (1988).
Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405 (1987).
Jorgensen R: Silencing of plant genes by homogous transgenes. AgBiotech News Inf 4: 265N-273N (1991).
Jorgensen R: The germinal inheritance of epigenetic information in plants. Phil Trans R Soc Lond B 339: 173–181 (1993).
Klein TM, Gradziel T, Fromm ME, Sanford JC: Factors influencing gene delivery intoZea mays cells by high-velocity microprojectiles. Bio/Technology 6: 559–563 (1988).
Klein TM, Kornstein L, Sanford JC: Genetic transformation of maize by particle bombardment. Plant Physiol 91: 440–444 (1989).
Koziel MG, Beland GL, Bowman C, Carozzi NB, Crenshaw R, Crossland L, Dawson J, Desai N, Hill M, Kadwell S, Launis K, Lewis K, Maddox D, McPherson K, Meghji MR, Merlin E, Rhodes R, Warren GW, Wright M, Evola SV: Field performance of elite transgenic maize plants expressing an insecticidal protein derived fromBacillus thuringiensis. Bio/Technology 11: 194–200 (1993).
Mettler IJ: A simple and rapid method for minipreparation of DNA from tissue cultured plant cells. Plant Mol Biol Rep 5: 346–349 (1987).
Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).
Omirulleh S, Abraham M, Golovkin M, Stefanov I, Karabaev MK, Mustardy L, Morocz S, Dudits D: Activity of a chimeric promoter with the doubled CaMV 35S enhancer dlement in protoplast-derived cells and transgenic plants in maize. Plant Mol Biol 21: 415–428 (1993).
Potrykus I, Paskowski J, Saul MW, Petruske J, Shillito R: Molecular and general genetics of a hybrid foreign gene introduced into tobacco by direct gene transfer. Mol Gen Genet 199: 169–177 (1985).
Riggs CD, Bates GW: Stable transformation of tobacco by electroporation: evidence for plasmid concatenation. Proc Natl Acad Sci USA 83: 5602–5606 (1986).
Rogers SO, Bendich AJ: Extraction of DNA from plant tissues. In: Gelvin SB, Schilperoort R (eds) Plant Molecular Biology Manual, pp. A6 1–10. Kluwer Academic Publishers, Dordrecht, Netherlands (1988).
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).
Shillito RD, Carswell GK, Johnson CM, DiMaio JJ, Harms C: Regeneration of fertile plants from protoplasts of elite inbred maize. Bio Technology 7: 581–587 (1989).
Spencer TM, O'Brien JV, Start WG, Adams TR, Gordon-Kamm WJ, lemaux PG: Segregation of transgenes in maize. Plant Mol Biol 18: 201–210 (1992).
Tomes DT, Weissinger AK, Ross M, Higgins R, Drummond BJ, Schaaf S, Malone-Schoneberg J, Staebel M, Flynn P, Anderson J, Howard J: Transgenic tobacco plants and their progeny derived by microprojectile bombardment of tobacco leaves. Plant Mol Biol 14: 261–268 (1990).
Torne JM, Santos MA, Blanco JL: Methods of obtaining maize totipotent tissues. II Atrophic tissue culture. Plant Sci Lett 38: 317–325 (1984).
Walters DA, Vetsch CS, Potts DE, Lundquist RC: Transformation and inheritance of a hygromycin phosphotransferase gene in maize plants. Plant Mol Biol 18: 189–200 (1992).
Wilson HM, Bullock WP, Dunwell JM, Ellis JR, Frame B, RegisterIII JC, Thompson JA: Maize transformation. In: VanMontague M, Wang K, Herrera-Estrella A (eds) Transformation of Plants and Soil Microorganisms. Cambridge University Press, Cambridge, UK (1994).
Wohlleben W, Arnold W, Broer I, Hillemann D, Strauch E, Puhler A: A nucleotide sequence of the phosphinothricin N-acetyltransferase gene fromStreptomyces viridochromogenes Tu494 and its expression inNicotiana tabacum. Gene 70: 25–37 (1988).
Wright SY, Suner M-M, Bell PJ, Vaudin M, Greenland AJ: Isolation and characterization of male flower cDNAs from maize. Plant J 3: 41–49 (1993).
Yenofsky RL, Fine M, Pellow JW: A mutant neomycin phosphotransferase II gene reduces the resistance of transformants to antibiotic selection pressure. Proc Natl Acad Sci USA 87: 3435–349 (1990).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Register, J.C., Peterson, D.J., Bell, P.J. et al. Structure and function of selectable and non-selectable transgenes in maize after introduction by particle bombardment. Plant Mol Biol 25, 951–961 (1994). https://doi.org/10.1007/BF00014669
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00014669