Abstract
Stable transformation of perennial ryegrass (Lolium perenne L.) was achieved by biolistic bombardment of a non embryogenic cell suspension culture, using the hpt and gusA gene. The transformation yielded on the average 5 callus lines per bombardment (1.4×106 cells). Stable integration of the genes into the plant genome was demonstrated by Southern analysis of DNA, isolated from hygromycin-resistant callus lines. The gusA reporter gene, which was regulated by the constitutive promoter of the rice gene GOS2, was expressed in both transient and stable transformation assays, indicating that this promoter is suitable for expression of a transferred gene in perennial ryegrass. Long-term GUS expression was observed in ca. 40% of the callus lines, whereas the other callus lines showed instability after 6 months and 1 year of culture.
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References
Bower R, Birch RG: Transgenic sugarcane plants via microprojectile bombardment. Plant J 2: 409–416 (1992).
Christou P, Ford TL, Kofron M: Production of transgenic rice (Oryza sativa L.) plants from agronomically important Indica and Japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zygotic embryos. Bio/technology 9: 957–962 (1991).
Creemers-Molenaar J, van derValk P, Loeffen JPM, Zaal MACM: Plant regeneration from suspension cultures and protoplasts of Lolium perenne L. Plant Sci 63: 167–176 (1989).
dePater BS, van derMark F, Rueb S, Katagiri F, Chua NH, Schilperoort RA, Hensgens LAM: The promoter of the rice gene GOS2 is active in various different monocot tissues and binds rice nuclear factor ASF-1. Plant J 2: 837–844 (1992).
Feinberg AP, Vogelstein B: A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13 (1983).
Finer JJ, McMullen MD: Transformation of cotton (Gossypium hirsutum L.) via particle bombardment. Plant Cell Rep. 8: 586–589 (1990).
Fitch MMM, Manshardt RM, Gonsalves D, Slightom JL, Sanford JC: Stable transformation of papaya via microprojectile bombardment. Plant Cell Rep 9: 189–194 (1990).
Fujiwara T, Lessard PA, Beachy RN: Inactivation of the nopaline synthase gene by double transformation: reactivation by segregation of the induced DNA. Plant Cell Rep. 12: 133–138 (1993).
Gordon-Kamm WJ, Spencer TM, Mangano ML, Adams TR, Daines RJ, Start WG, O'Brien JV, Chambers SA, Adams WRJr, 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).
Gritz L, Davies J: Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phototransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae. Gene 25: 179–188 (1983).
Hensgens LAM, Fornerod MWJ, Rueb S, Winkler AA, van derVeen S, Schilperoort RA: Translation controls the expression level of a chimeric reporter gene. Plant Mol Biol 20: 921–928 (1992).
Hensgens LAM, Rueb S, de Bakker EPHM, van Os-Ruygrok EP, Rueb S, van der Mark F, van der Maas HM, van der Veen S, Kooman-Gersmann M, 't Hart L, Schilperoort RA: Transient and stable expression of gusA fusions with rice genes in rice, barley and perennial expression. Plant Mol Biol, in press.
Jefferson RA, Kavanagh TA, Bevan MW: GUS fusions: β-glucuronidase as sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).
Klein TM, Wolf ED, Wu R, Sanford JC: High velocity microprojectiles for delivering nucleic acids into living cells. Nature 327: 70–73 (1987).
Klein TM, Kornstein L, Sanford JC, Fromm ME: Genetic transformation of maize cells by particle bombardment. Plant Physiol 91: 440–444 (1989).
Linn F, Heidmann I, Saedler H, Meyer P: Epigenetic changes in the expression of the maize A1 gene in Petunia hybrida: role of numbers of integrated gene copies and state of methylation. Mol Gen Genet 222: 329–336 (1990).
Matzke MA, Primig M, Trnovsky J, Matzke AJM: Reversible methylation and inactivation of marker genes in sequentially transformed tobacco plants. EMBO J 8: 643–649 (1989).
McCabe DE, Swain WF, Martinell BJ, Christou P: Stable transformation of soybean (Glycine max) by particle acceleration. Bio/technology 6: 923–926 (1988).
Meijer EGM, Schilperoort RA, Rueb S, vanOs-Ruygrok EP, Hensgens LAM: Transgenic rice cell lines and plants: expression of transferred chimeric genes. Plant Mol Biol 16: 807–820 (1991).
Mettler IJ: A simple and rapid method for minipreparation of DNA from tissue cultured plant cells. Plant Mol Biol Rep 5: 346–349 (1987).
Rueb S, Hensgens LAM: Improved histochemical staining for β-D-glucuronidase activity in monocotyledonous plants. Rice Genet Newsl 6: 168–169 (1989).
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).
Potrykus IP, Saul MW, Petruska J, Paszkowski J, Shillito RD: Direct gene transfer to cells of a gramineous monocol. Mol Gen Genet 199: 183–188 (1985).
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van der Maas, H.M., de Jong, E.R., Rueb, S. et al. Stable transformation and long-term expression of the gusA reporter gene in callus lines of perennial ryegrass (Lolium perenne L.). Plant Mol Biol 24, 401–405 (1994). https://doi.org/10.1007/BF00020178
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DOI: https://doi.org/10.1007/BF00020178