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Transient gene expression in aleurone protoplasts isolated from developing caryopses of barley and wheat

Plant Molecular Biology volume 13, pages 21–29 (1989)Cite this article

Abstract

Methods have been developed for the isolation of aleurone protoplasts from developing caryopses of Hordeum vulgare and Triticum aestivum in order to study transient expression of introduced genes. Chimaeric gene constructs were introduced into aleurone protoplasts by polyethylene glycol (PEG). Transient expression directed by the 35S promoter from cauliflower mosaic virus (CaMV) of the reporter gene encoding chloramphenicol acetyl transferase (CAT) was detected in aleurone protoplasts from developing barley and wheat grains. Using a similar construct, CAT activity increased when the alcohol dehydrogenase intron 1 fragment from maize was ligated between the 35S promoter and the CAT coding region. The demonstration of transient expression in protoplasts from developing aleurone layers indicates that they may be useful for investigating tissue and developmental control of genes coding for cereal seed proteins.

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References

  1. Callis J, Fromm M, Walbot V: Introns increase gene expression in cultured maize cells. Genes and Devel 1: 1183–1200 (1987).

    Google Scholar 

  2. Colot V, Robert L, Kavanagh T, Bevan M, Thompson R: Localization of sequences in wheat endosperm protein genes which confer tissue-specific expression in tobacco. EMBO J 6: 3559–3564 (1987).

    PubMed  Google Scholar 

  3. Fromm M, Callis J, Taylor LP, Walbot V: Electroporation of DNA and RNA into plant protoplasts. Meth Enzymol 153: 351–366 (1987).

    Google Scholar 

  4. Fromm M, Taylor LP, Walbot V: Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci USA 82: 5822–5828 (1985).

    Google Scholar 

  5. Gorman CM, Moffat LF, Howard BH: Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol 2: 1044–1051 (1982).

    PubMed  Google Scholar 

  6. Hauptmann RM, Ozias-Akins P, Vasil V, Tabaeizadeh Z, Rogers SG, Horsch RB, Vasil IK, Fraley RT: Transient expression of electroporated DNA in monocotyledonous and dicotyledonous species. Plant Cell Rep 6: 265–270 (1987).

    Article  Google Scholar 

  7. Hooley R: Protoplasts isolated from aleurone layers of wild oat (Avena fatua L.) exhibit the classic response to gibberellic acid. Planta 154: 29–40 (1982).

    Google Scholar 

  8. Jacobsen JV, Zwar JA and Chandler PM: Gibberellic-acid-responsive protoplasts from mature aleurone of Himalaya barley. Planta 163: 430–438 (1985).

    Google Scholar 

  9. Junker B, Zimney J, Lührs R, Lörz H: Transient expression of chimaeric genes in dividing and non-dividing cereal protoplasts after PEG-induced DNA uptake. Plant Cell Rep 6: 329–332 (1987).

    Article  Google Scholar 

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

    Google Scholar 

  11. Kreis M, Williamson MS, Forde J, Schmutz D, Clark J, Buxton B, Pywell J, Marris C, Henderson J, Harris N, Shewry PR, Forde BG, Miflin BJ: Differential gene expression in the developing barley endosperm. Phil Trans R Soc London B 314: 355–365 (1986).

    Google Scholar 

  12. Lindsey K, Jones MGK: Transient gene expression in electroporated protoplasts and intact cells of sugar been. Plant Mol Biol 10: 43–52 (1987).

    Google Scholar 

  13. Marcotte WRJr., Bayley CC, Quatrano RS: Regulation of a wheat promoter by abscisic acid in rice protoplasts. Nature 335: 454–457 (1988).

    Article  Google Scholar 

  14. Marris C, Gallois P, Copley J, Kreis M: The 5′ flanking region of a barley B hordein gene controls tissue and developmental specific CAT expression in tobacco plants. Plant Mol Biol 10: 359–366 (1988).

    Google Scholar 

  15. Negrutiu I, Shillito R, Potrykus I, Biasini G, Sala F: Hybrid genes in the analysis of transformation conditions I. Setting up a simple method for direct gene transfer in plant protoplasts. Plant Mol Biol 8: 363–373 (1987).

    Google Scholar 

  16. Norman HA, Black M, Chapman JM: The induction of sensitivity to gibberellin in aleurone tissue of developing wheat grains. Planta 158: 264–271 (1983).

    Google Scholar 

  17. Ou-Lee T-M, Turgeon R, Wu R: Expression of a foreign gene linked to either a plant-virus or a Drosophila promoter, after electroporation of protoplasts of rice, wheat, and sorghum. Proc Natl Acad Sci USA 83: 6815–6819 (1986).

    Google Scholar 

  18. Schernthaner JP, Matzke MA, Matzke AJM: Endosperm-specific activity of a zein gene promoter in transgenic tobacco plants. EMBO J 7: 1249–1255 (1988).

    Google Scholar 

  19. Walker JC, Howard EA, Dennis ES, Peacock WJ: DNA sequences required for anaerobic expression of the maize alcohol dehydrogenase 1 gene. Proc Natl Acad Sci USA 84: 6624–6628 (1987).

    Google Scholar 

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Affiliations

  1. Rothamsted Experimental Station, AFRC Institute of Arable Crops Research, AL5 2JQ, Harpenden, Herts, U.K.

    Bruce Lee, Kate Murdoch, Jennifer Topping, Martin Kreis & Michael G. K. Jones

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  1. Bruce Lee
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  2. Kate Murdoch
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  3. Jennifer Topping
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  4. Martin Kreis
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  5. Michael G. K. Jones
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Lee, B., Murdoch, K., Topping, J. et al. Transient gene expression in aleurone protoplasts isolated from developing caryopses of barley and wheat. Plant Mol Biol 13, 21–29 (1989). https://doi.org/10.1007/BF00027332

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  • DOI: https://doi.org/10.1007/BF00027332

Key words

  • aleurone
  • barley
  • protoplasts
  • transient expression
  • wheat