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Genetic Manipulation of Condensed Tannins in Higher Plants1
II. Analysis of Birdsfoot Trefoil Plants Harboring Antisense Dihydroflavonol Reductase Constructs

Mark P. Robbins*, Adrian D. Bavage2, Catherine Strudwicke, and Phillip Morris

Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, Ceredigion SY23 3EB, United Kingdom

We have produced and analyzed transgenic birdsfoot trefoil (Lotus corniculatus L.) plants harboring antisense dihydroflavonol reductase (AS-DFR) sequences. In initial experiments the effect of introducing three different antisense Antirrhinum majus L. DFR constructs into a single recipient genotype (S50) was assessed. There were no obvious effects on plant biomass, but levels of condensed tannins showed a statistical reduction in leaf, stem, and root tissues of some of the antisense lines. Transformation events were also found, which resulted in increased levels of condensed tannins. In subsequent experiments a detailed study of AS-DFR phenotypes was carried out in genotype S33 using pMAJ2 (an antisense construct comprising the 5' half of the A. majus cDNA). In this case, reduced tannin levels were found in leaf and stem tissues and in juvenile shoot tissues. Analysis of soluble flavonoids and isoflavonoids in tannin down-regulated shoot tissues indicated few obvious default products. When two S33 AS-DFR lines were outcrossed, there was an underrepresentation of transgene sequences in progeny plants and no examples of inheritance of an antisense phenotype were observed. To our knowledge, this is the first report of the genetic manipulation of condensed tannin biosynthesis in higher plants.

1   The Institute of Grassland and Environmental Research is grant funded by the Biotechnology and Biological Sciences Research Council (BBSRC). This work was supported by the BBSRC Plant Molecular Biology Initiative (parts 1 and 2).
2   Present address: John Innes Research Centre, Colney Lane, Norwich NR4 7UH, UK.
*   Corresponding author; e-mail mark.robbins{at}bbsrc.ac.uk; fax 44-1970-828357.

Plant Physiol. (1998) 116: 1133-1144
Copyright Clearance Center:   0032-0889/98/116/1133/12
© 1998 American Society of Plant Physiologists




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