Expression of fission yeast cdc25 driven by the wheat ADP-glucose pyrophosphorylase large subunit promoter reduces pollen viability and prevents transmission of the transgene in wheat
Article first published online: 17 JAN 2005
Volume 166, Issue 1, pages 185–192, April 2005
How to Cite
Chrimes, D., Rogers, H. J., Francis, D., Jones, H. D. and Ainsworth, C. (2005), Expression of fission yeast cdc25 driven by the wheat ADP-glucose pyrophosphorylase large subunit promoter reduces pollen viability and prevents transmission of the transgene in wheat. New Phytologist, 166: 185–192. doi: 10.1111/j.1469-8137.2004.01299.x
- Issue published online: 17 JAN 2005
- Article first published online: 17 JAN 2005
- Received: 6 October 2004 Accepted: 26 October 2004
- ADP-glucose pyrophosphoryase;
- fission yeast;
- transgenic wheat (Triticum aestivum)
- • Cell number was to be measured in wheat (Triticum aestivum) endosperm expressing Spcdc25 (a fission yeast cell-cycle regulator) controlled by a supposedly endosperm-specific promoter, AGP2 (from the large subunit of ADP glucose pyrophosphorylase).
- • Wheat was transformed by biolistics either with AGP2::GUS or AGP2::Spcdc25. PCR and RT–PCR checked integration and expression of the transgene, respectively.
- • In cv. Chinese Spring, AGP2::GUS was unexpectedly expressed in carpels and pollen, as well as endosperm. In cv. Cadenza, three AGP2::Spcdc25 plants, AGP2::Spcdc25.1, .2 and .3, were generated. Spcdc25 expression was detected in mature leaves of AGP2::Spcdc25.1/.3 which exhibited abnormal spikes, 50% pollen viability and low seed set per plant; both were small compared with the nonexpressing and normal AGP2::Spcdc25.2. Spcdc25 was not transmitted to the T1 in AGP2::Spcdc25.1 or .3, which developed normally. Spcdc25 was PCR-positive in AGP2::Spcdc25.2, using primers for a central portion, but not with primers for the 5′ end, of the ORF, indicating a rearrangement; Spcdc25 was not expressed in either T0 or T1.
- • The AGP2 promoter is not tissue-specific and Spcdc25 expression disrupted reproduction.