Abstract
Seven days after anthesis, the shoot apical meristem of immature embryos of wheat (Triticum aestivum L.) is not yet covered by the coleoptile or leaf primordia and provides an optimal target for ballistic micro-targeting. Gold particles 1.2 μm in diameter at a concentration of 5·105 particles per μl and propelled by 110-bar nitrogen penetrated up to four cell layers into embryo apical meristems but produced no deleterious effects on germination. The use of diaphragms with internal diameters of 100 or 200 μm restricted bombardment to meristem cells or also included surrounding tissues, respectively. The results of transient-expression experiments indicated successful delivery of foreign DNA into meristem cells. Cells of the central zone of the meristem or pro-meristem transiently expressed foreign genes driven by the Cauliflower mosaic virus (CaMV) 35S and rice actin1-D constitutive promoters. Partial plasmolysis before bombardment and slow recovery of normal turgor pressure increased transient-expression frequencies. Meristem cells transiently expressed foreign genes at frequencies 10-fold less than surrounding tissues under identical conditions. Transgenic sectors were observed in both coleoptiles and leaf primordia.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- CaMV:
-
cauliflower mosaic virus
- DPA:
-
days post anthesis
- GUS:
-
β-glucuronidase
- MSE:
-
Murashige Skoog medium
References
Avery Jr., G.S. (1930) Comparative anatomy and morphology of embryos and seedlings of maize, oats, and wheat. Bot. Gazette 89, 1–39
Bidney, D., Scelonge, C., Martich, J., Burrus, M., Sims, L., Huffman, G. (1992) Microprojectile bombardment of plant tissues increases transformation frequency by Agrobacterium tumefaciens. Plant Mol. Biol. 18, 301–313
Bruns, E. (1892) Der Grasembryo. Flora 76, 1–33
Christou, P. (1992) Genetic transformation of crop plants using microprojectile bombardment. Plant J. 2, 275–281
Christou, P., Swain, W.F., Yang, N.S., McCabe, D.E. (1989) Inheritance and expression of foreign genes in transgenic soybean plants. Proc. Natl. Acad. Sci. USA 86, 7500–7504
Chen, D.F., Dale, P.J. (1992) A comparison of methods for delivering DNA to wheat: the application of wheat dwarf virus DNA to seeds with exposed apical meristems. Transgenic Res. 1, 93–100
Cone, K., Burr, F., Burr, B. (1986) Molecular analysis of the maize anthocyanin regulatory locus C1. Proc. Natl. Acad. Sci. USA 83, 9631–9635
Goff, S.A., Klein, T., Roth, B.A., Fromm, M., Cone, K., Radicella, J.P., Chandler, V. (1990) Transactivation of anthocyanin biosynthetic genes following transfer of B regulatory genes into maize tissues. EMBO J. 9, 2517–2522
Hermann, R., Müller, M. (1991) High resolution biological scanning electron microscopy: a comparative study of low temperature metal coating techniques. J. Electr. Microsc. Techn. 18, 440–449
Hussey, G., Johnson, R.D., Warren, S. (1989) Transformation of meristematic cells in the shoot apex of cultured pea shoots by Agrobacterium tumefaciens and A. rhizogenes. Protoplasma 148, 101–105
Jefferson, R.A., Kavanagh, T.A., Bevan, M.W. (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6, 3901–3907
Jegla, D.E., Sussex, I.M. (1989) Cell lineage patterns in the shoot meristem of the sunflower embryo in the dry seed. Dev. Biol. 131, 215–225
Johansen, D.A. (1935) Dehydration and infiltration. Science 82, 253–254
Klein, T.M., Roth, B.A., Fromm, M.E. (1989) Advances in direct gene transfer into cereals. In: Genetic engineering, vol. 2, pp. 13–31, Nijkamp, H.J.J., van der Plas, L.H.W., van Aartrijk, J., eds. Plenum Publishing Corporation
Klein, T.M., Goff, S.A., Roth, B.A., Fromm, M.E. (1990) Applications of the particle gun in plant biology. In: Progress in plant cellular and molecular biology, pp. 56–66, Setlow, J.K., ed. Kluwer Academic Publishers, The Netherlands
Klein, T.M., Arenzen R., Lewis P.A., Fitzpatrick-McElligott, S. (1992) Transformation of microbes, plants and animals by particle bombardment. Bio/Technology 10, 286–291
Maniatis, T., Fritsch, E.F., Sambrook, J. (1982) Molecular cloning (A laboratory manual). Cold Spring Harbor Laboratory, New York
McCabe, D.E., Swain, W.F., Martinell, B.J., Christou, P. (1988) Stable transformation of soybean (Glycine max) by particle acceleration. Bio/Technology 6, 923–926
McCall, M.A. (1934) Developmental anatomy and homologies in wheat. J. Agric. Res. 48, 283–321
McDaniel, C.N., Poethig, S.R. (1988) Cell-lineage patterns in the shoot apical meristem of the germinating maize embryo. Planta 175, 13–22
McElroy, D., Zhang, W., Cao, J., Wu, R. (1990) Isolation of an efficient actin promoter for use in rice transformation. Plant Cell 2, 163–171
Medford, J.I. (1992) Vegetative apical meristems. Plant Cell 4, 1029–1039
Mendel, R.R., Müller, B., Schulze, J., Kolesnikov, V., Zelenin, A. (1989) Delivery of foreign genes to intact barley cells by highvelocity microprojectiles. Theor. Appl. Genet. 78, 31–34
Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–497
Nörner, C. (1881) Beitrag zur Embryoentwicklung der Gramineen. Flora W, 257–285
Perl, A., Wess, H., Blumenthal, A., Galili, G., Galun, E. (1992) Improvement of plant regeneration and GUS expression in scutellar wheat calli by optimization of culture conditions and DNA-microprojectile delivery procedures. Mol. Gen. Genet. 235, 279–284
Potrykus, I. (1990) Gene transfer to cereals: an assessment. Bio/Technology 8, 535–542
Roth, I. (1955) Zur morphologischen Deutung des Grasembryos und verwandter Embryotypen. Flora 142, 564–600
Sautter, C., Waldner, H., Neuhaus-Url, G., Galli, A., Neuhaus, G., Potrykus, I. (1991) Micro-targeting: high efficiency gene transfer using a novel approach for the acceleration of micro-projectiles. Bio/Technology 9, 1080–1085
Sussex, I.M. (1989) Developmental programming of the shoot meristem. Cell 56, 225–229
Steeves, T.A., Sussex, I.M. (1989) Patterns in plant development. Cambridge University Press
Sussex, I.M., Steeves, T.A. (1967) Apical initials and the concept of pro-meristem. Phytomorphology 17, 387–391
van Tieghem, P. (1872) Observations anatomiques sur le cotyledon des graminées. Ann. Sci. Nat. Bot. 5, 236–276
Vasil, V., Castillo, A.M., Fromm, M.E., Vasil, I.K. (1992) Herbicide resistant fertile transgenic wheat plants obtained by microprojectile bombardment of regenerable embryogenic callus. Bio/Technology 10, 667–674
Ye, G.N., Daniell, H., Sanford, J.C. (1990) Optimization of delivery of foreign DNA into higher-plant chloroplasts. Plant Mol. Biol. 15, 809–819
Author information
Authors and Affiliations
Additional information
We are indebted to Elke Fenner for invaluable technical assistance, to Ted Klein (DuPont Agric. Products, Wilmington, Del., USA) and Ray Wu (Section of Biochemistry, Cornell University, Ithaca, N.Y., USA) for providing the plasmids pBC17 and pAct1-D, respectively, and to Pat King for language editing of the manuscript.
Rights and permissions
About this article
Cite this article
Iglesias, V.A., Gisel, A., Bilang, R. et al. Transient expression of visible marker genes in meristem cells of wheat embryos after ballistic micro-targeting. Planta 192, 84–91 (1993). https://doi.org/10.1007/BF00198696
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00198696