Summary
We have used microprojectile bombardment of tobacco pollen to study the DNA sequences involved in the expression of pollen-expressed genes. Promoter-reporter gene fusions constructed with the promoters of three different pollen-expressed genes from tomato (LAT52, LAT56, and LAT59) and either theβ-glucuronidase or luciferase reporter genes were assayed by bombarding hydrated tobacco pollen with the gene constructs precipitated onto tungsten microprojectiles. Reporter gene expression can be assayed within 30 min, with the maximal level of expression between 6 and 12 h after bombardment. By constructing and assaying promoter deletion derivatives, we have been able to delimit regions of the promoters that are necessary for high level expression in pollen. We also demonstrate that results with this transient expression system parallel the expression levels seen in pollen from stably transformed transgenic plants. The microprojectile bombardment assay can be used to rapidly test constructs for pollen expression beforeAgrobacterium-mediated plant transformation. Furthermore, it may be possible to adapt the microprojectile bombardment technique to achieve stable transformation of pollen.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Booy, G.; Krens, F. A.; Huizing, H. J. Attempted pollen-mediated transformation of maize. J. Plant Physiol., 135:319–324; 1989.
Bruce, W. B.; Christensen, A. H.; Klein, T., et al. Photoregulation of a phytochrome gene promoter from oat transferred into rice by particle bombardment. Proc. Natl. Acad. Sci. USA 86:9692–9696; 1989.
Hamilton, D. A.; Bashe, D. M.; Stinson, J. R., et al. Characterization of a pollen-specific genomic clone from maize. Sex Plant Reprod. 2:208–212; 1989.
Hanson, D. D.; Hamilton, D. A.; Travis, J. L., et al. Characterization of a pollen-specific cDNA clone fromZea mays and its expression. Plant Cell 1:173–179; 1989.
Jefferson, R. A.; Kavanagh, T. A.; Bevan, M. W.β-Glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6:3901–3907; 1987.
Johnston, S. A.; Riedy, M.; De Vit, M. J., et al. Biolistic transformation of animal tissue. In Vitro Cell. Dev. Biol. 27P:Add when known; 1991.
Klein, T. M.; Gradziel, T.; Fromm, M. E., et al. Factors influencing gene delivery intoZea mays cells by high-velocity microprojectiles. Bio/technology 6:559–566; 1988.
Loewus, F. A.; Baldi, B. G.; Franceschi, V. R., et al. Pollen sporoplasts: dissolution of pollen walls. Plant Physiol. 78:652–654; 1985.
Mascarenhas, J. P. The male gametophyte of flowering plants. Plant Cell 1:657–664; 1989.
McCormick, S. Tomato transformation withAgrobacterium tumefaciens. In: Lindsey, K., ed. Plant tissue culture manual: fundamentals and applications. Dordrecht, The Netherlands: Kluwer Academic Publishers, in press; 1990.
Ow, D. W.; Wood, K. V.; DeLuca, M., et al. Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Science 234:856–859; 1986.
Roeckel, P.; Heinzmann, P.; DuBois, M., et al. Attempts to transformZea mays via pollen grains. Effect of pollen and stigma nuclease activities. Sex Plant Reprod. 1:156–163; 1988.
Russell, J. A.; Roy, M. K.; Sanford, J. C. Cell injury as a limiting factor in stable biolistic plant transformation. In Vitro Cell. Dev. Biol. 26:43A; 1990.
Twell, D.; Wing, R. A.; Yamaguchi, J., et al. Isolation and expression of an anther-specific gene from tomato. Mol. Gen. Genet. 217:240–245; 1989a.
Twell, D.; Klein, T. M.; Fromm, M. E., et al. Transient expression of chimeric genes delivered into pollen by microprojectile bombardment. Plant Physiol. 91:1270–1274; 1989b.
Twell, D.; Yamaguchi, J.; McCormick, S. Pollen-specific gene expression in transgenic plants: coordinate regulation of two different tomato gene promoters during microsporogenesis. Development 109:705–713; 1990.
Twell, D.; Klein, T. M.; McCormick, S. Transformation of pollen by particle bombardment. In: Lindsey, K., ed. Plant tissue culture manual: fundamentals and applications. Dordrecht, The Netherlands:Kluwer Academic Publishers, in press; 1990.
Ursin, V. M.; Yamaguchi, J.; McCormick, S. Gametophytic and sporophytic expression of anther-specific genes in developing tomato anthers. Plant Cell 1:727–736; 1989.
van Tunen, A. J.; Hartman, S. A.; Mur, L. A., et al. Regulation of chalcone isomerase (CHI) gene expression inPetunia hybrida: the use of alternative promoters in corolla, anthers and pollen. Plant. Mol. Biol. 12:539–551; 1989.
van Tunen, A. J.; Mur, L. A.; Brouns, G. S., et al. Pollen-and anther-specificchi promoters from petunia: tandem promoter regulation of thechiA gene. Plant Cell 2:393–401; 1990.
Wing, R. A. Yamaguchi, J.; Larabell, S. K., et al. Molecular and genetic characterization of two pollen-expressed genes that have sequence similarity to pectate lyases of the plant pathogenErwinia. Plant Mol. Biol. 14:17–28; 1989.
Author information
Authors and Affiliations
Additional information
Presented in the Session-in-Depth Genetic Transformation and Genetic Analysis Using Microprojectile Bombardment at the 41st Annual Meeting of the Tissue Culture Association, Houston, Texas, June 10–13, 1990.
This work was supported by the U.S. Department of Agriculture, Washington, DC, ARS CRIS 5335-22230-002-00D, and by the NSF Center for Plant Developmental Biology, UC-Berkeley, DIR-8719933.
Rights and permissions
About this article
Cite this article
McCormick, S., Yamaguchi, J. & Twell, D. Deletion analysis of pollen-expressed promoters. In Vitro Cell Dev Biol - Plant 27, 15–20 (1991). https://doi.org/10.1007/BF02632056
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02632056