Abstract
Many plant genetic engineering applications require spatial expression of genes which in turn depends upon the availability of specific promoters. In cereals, genetic modification of flowering and grain setting to influence yield and grain quality is of significant interest. PsEND1 is a pea promoter that displays expression in the epidermis, connective tissue, endothecium and middle layers during different stages of anther development. No homeologous sequence of this promoter or its coding sequence has been found in cereals. This present work aimed at the characterization of the pea PsEND1 promoter driving the expression of the gusA gene in transgenic wheat. Nine transgenic lines were produced by particle bombardment and analyzed for the expression of the gusA gene throughout development by histochemical GUS staining and by RT-PCR in vegetative and reproductive tissues and organs. Expression of the gusA gene was first detected during pollen development, in microspores at binucleate stage. Activity of the gusA gene was also found in mature pollen, after anthesis. Following pollen grain germination, expression of the gusA gene was seen from an early stage of pollen tube formation until advanced stages, approaching the ovary. No further expression of the gusA gene was detected after fertilization, nor during seed development. The results reported here show that the PsEND1 promoter is functional in wheat and its patterns of expression may be of interest for the application of genetic modification in wheat breeding.
This is a preview of subscription content, log in via an institution to check access.
References
Barcelo P, Lazzeri PA (1995) Transformation of cereals by microprojectile bombardment of immature inflorescence and scutellum tissues. In: Jones H (ed) Methods in molecular biology: plant gene transfer and expression protocols. Humana Press Inc., Totowa, NJ, pp 113–123
Barro F, Cannell ME, Lazzeri PA, Barcelo P (1998) The Influence of auxins on transformation of wheat and tritordeum and analysis of transgene integration patterns in transformants. Theor Appl Genet 97:684–695
Christensen AH, Quail PH (1996) Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants. Transgenic Res 5:213–218
Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158
Gomez M, Beltran JP, Cañas L (2004) The pea END1 promoter drives anther-specific gene expression in different plant species. Planta 219:967–981
Hunt LA, Vanderpoorten G, Pararajasingham S (1991) Postanthesis temperature effects on duration and rate of grain filling in some winter and spring wheats. Can J Plant Sci 71:609–617
Lamacchia C, Shewry PR, Di Fonzo N, Forsyth JL, Harris N, Lazzeri PA, Napier JA, Halford NG, Barcelo P (2001) Endosperm-specific activity of a storage protein gene promoter in transgenic wheat seed. J Exp Bot 52:243–250
Mascarenhas JP (1990) Gene activity during pollen development. Annu Rev Plant Physiol Plant Mol Biol 41:317–338
McCormick S (2004) Control of male gametophyte development. Plant Cell 16:S142–S153
Reynolds TL (1997) Pollen embryogenesis. Plant Mol Biol 33:1–10
Rogers HJ, Bate N, Combe J, Sullivan J, Sweetman J, Swan C, Lonsdale DM, Twell D (2001) Functional analysis of cis-regulatory elements within the promoter of the tobacco late pollen gene g10. Plant Mol Biol 45:577–585
Roque E, Gómez M, Ellul P, Wallbraun M, Madueño F, Beltrán J-P, Cañas L (2007) The PsEND1 promoter: a novel tool to produce genetically engineered male-sterile plants by early anther ablation. Plant Cell Rep 26:313–325
Schäffner AR, Sheen J (1991) Maize rbcS promoter activity depends on sequence elements not found in dicot rbcS promoters. Plant Cell 3:997–1012
Stacey J, Isaac PG (1994) Isolation of DNA from plants. In: Isaac PG (ed) Methods in molecular biology: protocols for nucleic acid analysis by nonradioactive probes. Humana Press Inc., Totowa, New Jersey, pp 9–15
Stoger E, Williams S, Keen D, Christou P (1999) Constitutive versus specific expression in transgenic wheat: temporal and spatial control. Transgenic Res 8:73–82
Stone PJ, Nicolas ME (1994) Wheat cultivars vary widely in their responses of grain-yield and quality to short periods of postanthesis heat-stress. Aus J Plant Physiol 21:887–900
Twell D, Yamaguchi J, Wing RA, Ushiba J, McCormick S (1991) Promoter analysis of genes that are coordinately expressed during pollen development reveals pollen-specific enhancer sequences and shared regulatory elements. Genes Dev 5:496–507
Ueng P, Galili G, Sapanara V, Goldbrough PB, Dube P, Beachy RN, Larkins BA (1988) Storage protein gene in petunia plants is not restricted to seeds. Plant Physiol 86:1281–1285
Wardlaw IF, Moncur L (1995) The response of wheat to high-temperature following anthesis. 1. The rate and duration of kernel filling. Aus J Plant Physiol 22:391–397
Zheng Z, Kawagoe Y, Xiao S, Li Z, Okita T, Hau TL, Lin A, Murai N (1993) 5′ distal and proximal cis-acting regulator elements are required for developmental control of a rice seed storage protein glutelin gene. Plant J 4:357–366
Acknowledgements
The authors acknowledge funding by the Spanish C.I.C.Y.T. (project AGL2007-65685-C02-01/AGR, and PTR1995-0979-OP-03-03). The authors thank to Dr. Paul Lazzeri (Agrasys, SL) for the reading of the manuscript. Technical assistance by Ana García is also acknowledged.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Pistón, F., García, C., de la Viña, G. et al. The pea PsEND1 promoter drives the expression of GUS in transgenic wheat at the binucleate microspore stage and during pollen tube development. Mol Breeding 21, 401–405 (2008). https://doi.org/10.1007/s11032-007-9133-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11032-007-9133-7