Abstract
Immature tobacco (Nicotiana tabacum L.) pollen has been isolated from anthers in three distinct stages of development, including the microspore stage. In in-vitro cultures, fully functional, mature pollen was obtained. In a germination medium, this pollen produced pollen tubes. After application to stigmas in situ, the in-vitro-matured pollen fertilized ovules, and seeds were produced. Genetic tests with seedlings obtained from pollinations with in-vitro-matured pollen from a transgenic plant revealed normal Mendelian segregation of two marker genes, the neomycin-phosphotransferase II gene and the nopaline-synthase gene. These results are of interest with respect to the control of self-incompatibility, cytoplasmic male sterility and pollen-allergen formation, and it offers an alternative route for gene transfer in those plants which cannot be regenerated in vitro.
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Abbreviations
- cms:
-
cytoplasmic male sterility; AMGLU, MS, M2S, MR26
- GK:
-
culture media, see Material and methods
References
Brewbaker, J.L., Kwack, B.H. (1963) The essential role of calcium ion in pollen germination and pollen tube growth. Am. J. Bot. 50, 859–865
Dunwell, J.M, Sunderland, N.J. (1974) Pollen ultrastructure in anther cultures of Nicotiana tabacum I. Early stages of culture. Exp. Bot. 25, 352–361
Heberle-Bors, E. (1984) Genotypic control of pollen embryogenesis in Nicotiana tabacum L. theor. Appl. Genet. 68, 475–479
Heberle-Bors, E., Charvat, B., Thompson, D., Schernthaner, J.P., Barta, A., Matzke, A.J.M., Matzke, M.A. (1988) Genetic analysis of T-DNA insertions into the tobacco genome. Plant Cell Rep., in press
Heberle-Bors, E., Reinert, J. (1980) Isolated pollen cultures and pollen dimorphism. Naturwissenschaften 67, 311
Horsch, R.B., Fry, J.E., Hoffman, N.L., Eichholtz, D., Rogers, S.G., Fraley, R.T. (1985) A simple and general method for transferring genes into plants. Science 227, 1229–1231
Ito, M., Stern, H. (1967) Studies of meiosis in vitro I. In vitro culture of meiotic cells. Dev. Biol. 16, 36–53
Kyo, M., Harada, H. (1986) Control of the development pathway of tobacco pollen in vitro. Planta 168, 427–432
Mascarenhas, J.P., Eisenberg, A., Stinson, J.S., Willing, R.P., Pé, M.E. (1985) Genes expressed during pollen development. In: Plant cell/cell interactions, pp. 19–23, Sussex, I., Ellingboe, A., Crouch, M., Malmberg, R., eds. Cold Spring Harbor Laboratory
Matzke, A.J.M., Matzke, M.A. (1986) A set of novel Ti plasmid-derived vectors for the production of transgenic plants. Plant Mol. Biol. 7, 357–365
Murashige, T., Skoog, E. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–497
Otten, E.A.B.M., Schilperoort, R.A. (1978) A rapid microscale method for the detection of lysopine and nopaline dehydrogenase activities. Biochem. Biophys. Acta 527, 497–500
Pareddy, D.R., Greyson, R.I., Walden, D.B. (1987) Fertilization and seed production with pollen from in-vitro-cultured maize tassels. Planta 170, 141–143
Sangwan, R.S., Sangwan-Norreel, B.S. (1987) Ultrastructural cytology of plastids in pollen grains of certain androgenic and nonandrogenic plants. Protoplasma 138, 11–22
Takegami, M.H., Yoshioka, M., Tanaka, I., Ito, M. (1981) Cltaracteristics of isolated microsporocytes from liliaceous plants for studies of the meiotic cell cycle in vitro. Plant Cell Physiol. 22, 1–10
Tanaka, I., Ito, M. (1980) Induction of typical cell division in isolated microspores of Lilium longiflorum and Tulipa gesneriana Plant Sci Lett 17, 279–285
Tupy, J. (1982) Alterations in polyadenylated RNA during pollen maturation and germination. Biol. Plant. 24, 331–340
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Moreno, R.M.B., Macke, F., Alwen, A. et al. In-situ seed production after pollination with in-vitro-matured, isolated pollen. Planta 176, 145–148 (1988). https://doi.org/10.1007/BF00392439
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DOI: https://doi.org/10.1007/BF00392439