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

D. Chrimes, H. J. Rogers, D. Francis, H. D. Jones, C. Ainsworth

Research output: Contribution to journalArticlepeer-review

16 Citations (SciVal)

Abstract

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 T-1 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 T-0 or T-1.

The AGP2 promoter is not tissue-specific and Spcdc25 expression disrupted reproduction.

Original languageEnglish
Pages (from-to)185-192
Number of pages8
JournalNew Phytologist
Volume166
Issue number1
DOIs
Publication statusPublished - Apr 2005

Keywords

  • ADP-glucose pyrophosphoryase
  • endosperm
  • fission yeast
  • Spcdc25
  • transgenic wheat (Triticum aestivum)
  • CELL-DIVISION
  • PROTEIN
  • KINASE
  • CYCLE
  • MITOSIS
  • PLANTS
  • GENES
  • TRANSFORMATION
  • PHOSPHATASE
  • FREQUENCY

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