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Transient expression of GUS and anthocyanin constructs in intact maize immature embryos following electroporation

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Abstract

Electroporation was used for the delivery and subsequent expression of GUS and anthocyanin reporter genes into intact maize immature embryos. The optimal conditions consisted of culturing immature embryos for 4 days on N6 1-100-25-Ag medium prior to electroporation (375 V/cm; 960 µF capacitance) in EPR buffer containing DNA and 0.07 M sodium glutamate at room temperature (22°C) after a 10 min heat shock at 37°C. Under these conditions, over 40 spots of GUS transient activity were observed per immature embryo. Transient gene expression after electroporation was further demonstrated using an anthocyanin construct, which is specific for expression in plant cells.

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Abbreviations

EPR:

electroporation buffer

GUS:

β-glucuronidase

2,4-D:

2,4-dichlorophenoxyacetic acid

References

  • Abdul-Baki AA, Saunders JA, Matthews BF & Pittarelli GW (1990) DNA uptake during electroporation of germinating pollen grains. Plant Sci. 70: 181–190

    Article  Google Scholar 

  • Armstrong CL & Green CE (1985) Establishment and maintenance of friable, embryogenic maize callus and the involvement ofl-proline. Planta 164: 207–214

    Article  Google Scholar 

  • Dekeyser RA, Claes B, De Rycke RMU, Habets ME, Van Montagu MC & Caplan AB (1990) Transient gene expression in intact and organized rice tissues. The Plant Cell 2: 591–602

    Article  PubMed  Google Scholar 

  • Fromm ME, Morrish F, Armstrong C, Williams R, Thomas J & Klein TM (1990) Inheritance and expression of chimeric genes in the progeny of transgenic maize plants. Biotechnol. 8: 833–839

    Article  Google Scholar 

  • Goff SA, Klein TM, Roth BA, Fromm ME, Cone KC, Radicella JP & Chandler VL (1990) Transactivation of anthocyanin biosynthetic genes following transfer of B regulatory genes into maize tissues. Embo J. 9: 2517–2522

    PubMed  Google Scholar 

  • Gordon-Kamm WJ, Spencer TM, Mangano ML, Adams TR, Daines RJ, Start WG, O'Brien JV, Chambers SA, Adams Jr. WR, Willetts NG, Rice TB, Mackey CJ, Krueger RW, Kausch AP & Lemaux PG (1990) Transformation of maize cells and regeneration of fertile transgenic plants. The Plant Cell 2: 603–618

    Article  PubMed  Google Scholar 

  • Jefferson RA, Kavanagh TA & Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6: 3901–3907

    PubMed  Google Scholar 

  • Kaeppler HF, Weining G, Somers DA, Rines HW & Cockburn AF (1990) Silicon carbide fiber-mediated DNA delivery into plant cells. Plant Cell Rep. 9: 415–418

    Article  Google Scholar 

  • Kartha KK, Chibbar RN, Georges F, Leung N, Caswell K, Kendall E & Qureshi J (1990) Transient expression of chloramphenicol acetyltransferase (CAT) gene in barley cell cultures and immature embryos through microprojectile bombardment. Plant Cell Rep. 8: 429–432

    Article  Google Scholar 

  • Klein TM, Wolf ED, Wu R & Sanford JC (1987) High-velocity microprojectiles for delivering nucleic acids into living cells. Nature 327: 70–73

    Article  Google Scholar 

  • Klein TM, Gradziel T, Fromme ME & Sanford JC (1988a) Factors influencing gene delivery intoZea mays cells by high-velocity microprojectiles. Biotechnol. 6: 559–563

    Article  Google Scholar 

  • Klein TM, Fromm M, Weissinger A, Tomes D, Schaff S, Sletten M & Sanford JC (1988b) Transfer of foreign genes into intact maize cells with high-velocity microprojectiles. Proc. Natl. Acad. Sci. 85: 4305–4309

    Google Scholar 

  • Li BJ, Xu XP, Shi HP & Ke XY (1991) Introduction of foreign genes into the seed embryo cells of rice by electroinjection and the regeneration of transgenic rice plants. Sci. China (Series B) 34: 923–931

    Google Scholar 

  • Matthews BF, Abdul-Baki AA & Saunders JA (1990) Expression of a foreign gene in electroporated pollen grains of tobacco. Sex. Plant Reprod. 3: 147–151

    Article  Google Scholar 

  • Oard JH, Paige DF, Simmonds JA & Gradziel TM (1990) Transient gene expression in maize, rice, and wheat cells using an airgun apparatus. Plant Physiol. 92: 334–339

    Google Scholar 

  • Senaratna T, McKersie BD, Kasha KJ & Procunier JD (1991) Direct DNA uptake during imbibition of dry cells. Plant Sci. 79: 223–228

    Article  Google Scholar 

  • Shillito RD, Saul MW, Paszkowski J, Muller M & Potrykus I (1985) High efficiency direct gene transfer to plants. Biotechnol. 3: 1099–1103

    Article  Google Scholar 

  • Songstad DD, Armstrong CL & Petersen WL (1991) AgNO3 increases type II callus production from immature embryos of maize inbred B73 and its derivatives. Plant Cell Rep. 9: 699–702

    Article  Google Scholar 

  • Tada Y, Sakamoto M & Fujimura T (1990) Efficients gene introduction into rice by electroporation and analysis of transgenic plants: use of electroporation buffer lacking chloride ions. Theor. Appl. Genet. 80: 475–480

    Article  Google Scholar 

  • Topfer R, Gronenborn B, Schell J & Steinbiss HH (1989) Uptake and transient expression of chimeric genes in seed-derived embryos. The Plant Cell 1: 133–139

    Article  PubMed  Google Scholar 

  • Van Wert SL & Saunders JA (1992) Reduction of nuclease activity released from germinating pollen under conditions used for pollen electrotransformation. Plant Sci. 84: 11–16

    Article  Google Scholar 

  • Wang YC, Klein TM, Fromm M, Cao J, Sanford JC & Wu R (1988) Transient expression of foreign genes in rice, wheat and soybean cells following particle bombardment. Plant Mol. Biol. 11: 433–439

    Article  Google Scholar 

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Author notes

  1. D. D. Songstad

    Present address: Pioneer Hi-Bred International, 7300 NW 62nd Ave., 50131, Johnston, Iowa, USA

Authors and Affiliations

  1. Monsanto Company, 700 Chesterfield Parkway North, 63198, St. Louis, MO, USA

    D. D. Songstad, F. G. Halaka, D. L. DeBoer, C. L. Armstrong, M. A. W. Hinchee, C. G. Ford-Santino, S. M. Brown, M. E. Fromm & R. B. Horsch

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  1. D. D. Songstad
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  2. F. G. Halaka
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  3. D. L. DeBoer
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  4. C. L. Armstrong
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  5. M. A. W. Hinchee
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  6. C. G. Ford-Santino
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  7. S. M. Brown
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  8. M. E. Fromm
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  9. R. B. Horsch
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Songstad, D.D., Halaka, F.G., DeBoer, D.L. et al. Transient expression of GUS and anthocyanin constructs in intact maize immature embryos following electroporation. Plant Cell Tiss Organ Cult 33, 195–201 (1993). https://doi.org/10.1007/BF01983234

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  • DOI: https://doi.org/10.1007/BF01983234

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