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Expression of a chimeric stilbene synthase gene in transgenic wheat lines
Authors:Fettig  Sebastian  Hess  Dieter
Institution:(1) IACR-Rothamsted, Harpenden, Herts, AL5 2JQ, U.K;(2) DuPont Wheat Transformation Laboratory, IACR-Rothamsted, Harpenden, Herts, AL5 2JQ, U.K;(3) IACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, Long Ashton, Bristol, BS41 9AF, U.K
Abstract:A chimeric stilbene synthase (sts)dashgene was transferred into wheat. Stilbene synthases play a role in the defence against fungal diseases in some plant species (e.g. groundnut or grapevine) by producing stilbenedashtype phytoalexins like resveratrol. Resveratrol is also claimed to have positive effects to human health. Embryogenic scutellar calli derived from immature embryos of the two commercial German spring wheat cultivars lsquoCombirsquo and lsquoHannorsquo were used as target tissue for codashtransformation by microprojectile delivery. The selectable marker/reporter gene constructs contained the bardashgene either driven by the ubiquitindashpromoter from maize (pAHC 25, also containing the uidAdashgene driven by the ubiquitindashpromoter), or by the actindashpromoter (pDM 302) from rice. The codashtransferred plasmid pStil 2 consisted of a grapevine stsdashcoding region driven by the ubiquitin promoter. Eight transgenic lsquoCombirsquo and one lsquoHannorsquo TOdashplant were obtained and, except one lsquoCombirsquo TOdashplant, found to be codashtransformants due to the integration of both the stsdashgene and the selectable marker or reporter genes. Expression of the stsdashgene was proven by RTdashPCR, and, for the first time, by detection of the stilbene synthase product resveratrol by HPLC and mass spectrometry. The stsdashgene was expressed in four of the seven transgenic lsquoCombirsquo T_odashplants. Two of the respective T1dashprogenies segregated in a Mendelian manner were still expressing the gene. Investigations into methylation of the stsdashgene showed that in three nondashexpressing progenies inactivation was paralleled by methylation.
Keywords:methylation  resveratrol  stilbene synthase  transgenic wheat
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