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Keywords:

  • ADP-glucose pyrophosphoryase;
  • endosperm;
  • fission yeast;
  • Spcdc25;
  • transgenic wheat (Triticum aestivum)

Summary

  • 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.