Expression of the BnmNAP subfamily of napin genes coincides with the induction of Brassica microspore embryogenesis |
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Authors: | Kim A. Boutilier María-Jesús Ginés Janice M. DeMoor Bin Huang Chris L. Baszczynski V. N. Iyer Brian L. Miki |
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Affiliation: | (1) Biology Department, University of Ottawa, K1N 6N5 Ottawa, Ontario, Canada;(2) Plant Research Centre, Agriculture Canada, K1A 0C6 Ottawa, Ontario, Canada;(3) Department of Biology, Carleton University, K1S 5B6 Ottawa, Ontario, Canada;(4) Plant Breeding Division, Pioneer Hi-Bred Production Ltd, L7G 4S7 Georgetown, Ontario, Canada;(5) Plant Breeding Division, Department of Biotechnology Research, Pioneer Hi-Bred International, Inc., 50131-0038 Iowa Johnston, Iowa, USA;(6) Present address: Department of Molecular Biology, Agricultural University of Wageningen, Dreijenlaan 3, 6703 HA Wageningen, Netherlands;(7) Present address: Experimental Oncology, London Regional Cancer Centre, N6A 4L6 London, Ontario, Canada |
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Abstract: | Brassica napus cv. Topas microspores can be diverted from pollen development toward haploid embryo formation in culture by subjecting them to a heat stress treatment. We show that this switch in developmental pathways is accompanied by the induction of high levels of napin seed storage protein gene expression. Changes in the plant growth or microspore culture conditions were not by themselves sufficient to induce napin gene expression. Specific members of the napin multigene family were cloned from a cDNA library prepared from microspores that had been induced to undergo embryogenesis. The majority of napin clones represented three members (BnmNAP2, BnmNAP3 and BnmNAP4) that, along with a previously isolated napin genomic clone (BngNAP1), constitute the highly conserved BnmNAP subfamily of napin genes. Both RNA gel blot analysis, using a subfamily-specific probe, and histochemical analysis of transgenic plants expressing a BngNAP1 promoter- -glucuronidase gene fusion demonstrated that the BnmNAP subfamily is expressed in embryogenic microspores as well as during subsequent stages of microsporic embryo development. |
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Keywords: | Brassica napus microspore embryogenesis napin |
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