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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Callis J, Fromm M, Walbot V: Introns increase gene expression in cultured maize cells. Genes and Devel 1: 1183–1200 (1987).
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).
Fromm M, Callis J, Taylor LP, Walbot V: Electroporation of DNA and RNA into plant protoplasts. Meth Enzymol 153: 351–366 (1987).
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).
Gorman CM, Moffat LF, Howard BH: Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol 2: 1044–1051 (1982).
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).
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).
Jacobsen JV, Zwar JA and Chandler PM: Gibberellic-acid-responsive protoplasts from mature aleurone of Himalaya barley. Planta 163: 430–438 (1985).
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).
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).
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).
Lindsey K, Jones MGK: Transient gene expression in electroporated protoplasts and intact cells of sugar been. Plant Mol Biol 10: 43–52 (1987).
Marcotte WRJr., Bayley CC, Quatrano RS: Regulation of a wheat promoter by abscisic acid in rice protoplasts. Nature 335: 454–457 (1988).
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).
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).
Norman HA, Black M, Chapman JM: The induction of sensitivity to gibberellin in aleurone tissue of developing wheat grains. Planta 158: 264–271 (1983).
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).
Schernthaner JP, Matzke MA, Matzke AJM: Endosperm-specific activity of a zein gene promoter in transgenic tobacco plants. EMBO J 7: 1249–1255 (1988).
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).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00027332