title: 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.
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.
authors.affiliation: School of Biosciences, Cardiff University, PO Box 915, Cardiff CF10 3TL, UK.
authors.forename: D
authors.initials: D
authors.lastname: Chrimes
authors.forename: H J
authors.initials: HJ
authors.lastname: Rogers
authors.forename: D
authors.initials: D
authors.lastname: Francis
authors.forename: H D
authors.initials: HD
authors.lastname: Jones
authors.forename: C
authors.initials: C
authors.lastname: Ainsworth
mesh_terms: D018506:Gene Expression Regulation, Plant; D051599:Glucose-1-Phosphate Adenylyltransferase; D009713:Nucleotidyltransferases / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D030821:Plants, Genetically Modified; D011058:Pollen / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012098:Reproduction / Q000235:genetics / Q000502:physiology; D014908:Triticum / Q000235:genetics / Q000502:physiology*; D020687:cdc25 Phosphatases / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*
chemical_list: D011993:Recombinant Fusion Proteins; D009713:Nucleotidyltransferases; D051599:Glucose-1-Phosphate Adenylyltransferase; D020687:cdc25 Phosphatases