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Published on November 10, 2006; 10.1104/pp.106.090886

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Received October 6, 2006
Accepted November 2, 2006

Ectopic Expression of a bHLH Gene Transactivates Parallel Pathways of Proanthocyanidin (PA) Biosynthesis. Structure, Expression Analysis and Genetic Control of LAR and ANR Genes in Lotus corniculatus

Francesco Paolocci *, Mark P. Robbins , Laura Madeo , Sergio Arcioni , Stefan Martens , and Francesco Damiani

National Research Council, Plant Genetics Institute, Perugia, CNR Via Madonna Alta, 130-06128 Perugia, Italy
Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, Ceredigion SY23 3EB, UK
Philipps Universität Marburg, Institut für Pharmazeutische Biologie, Deutschhausstr. 17A, 35037 Marburg/Lahn, Germany

* Corresponding author; email: francesco.paolocci{at}igv.cnr.it.

Proanthocyanidins (PAs) are plant secondary metabolites and are composed primarily of catechin and epicatechin units in higher plant species. Owing to the ability of PAs to bind reversibly with plant proteins to improve digestion and reduce bloat, engineering this pathway in leaves is a major goal for forage breeders. Here we report the cloning and expression analysis of anthocyanidin reductase (ANR) and leucoanthocyanidin 4-reductase (LAR), two genes encoding enzymes committed to epicatechin and catechin biosynthesis respectively, in Lotus corniculatus. We show the presence of two LAR gene families (LAR1 and LAR2) and that the steady state levels of ANR and LAR1 genes correlate with the levels of PAs in leaves of wild type and transgenic plants. Interestingly, ANR and LAR1, but not LAR2, genes produced active proteins following heterologous expression in E. coli and are affected by the same bHLH transcription factor that promotes PA accumulation in cells of palisade and spongy mesophyll. This study provides direct evidence that the same subclass of transcription factor can mediate the expression of the structural genes of both branches of PA biosynthesis.




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