Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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Published on November 11, 2009; 10.1104/pp.109.147322

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Received September 11, 2009
Accepted November 3, 2009

Biochemical and molecular analysis of pink tomatoes: deregulated expression of the gene encoding transcription factor SlMYB12 leads to pink tomato fruit colour

Ana-Rosa Ballester , Jos Molthoff , Ric de Vos , Bas te Lintel Hekkert , Diego Orzaez , Josefina-Patricia Fernandez-Moreno , Pasquale Tripodi , Silvana Grandillo , Cathie Martin , Jos Heldens , Marieke Ykema , Antonio Granell , and Arnaud Bovy *

Plant Research International, PO Box 16, 6700 AA Wageningen, The Netherlands; Centre for Biosystems Genomics, PO Box 98, 6700 PB Wageningen, The Netherlands; CNR-IGV, Institute of Plant Genetics, National Research Council, Via Universita 133, 80055 Portici, Italy; John Innes Centre, Norwich Research Park, Colney, Norwich, NR4 7UH, UK; Enza Zaden Research & Development B.V., P.O Box 7, 1600 AA Enkhuizen, The Netherlands; Instituto de Biologia Molecular y Celular de Plantas, Consejo Superior de Investigaciones Cientificas, Universidad Politecnica de Valencia, Camino de Vera s/n, 46022 Valencia, Spain

* Corresponding author; email: arnaud.bovy{at}wur.nl.

The colour of tomato fruit is mainly determined by carotenoids and flavonoids. Phenotypic analysis of an introgression line (IL) population derived from a cross between Solanum lycopersicum cv Moneyberg and the wild species Solanum chmielewskii revealed three ILs with a pink fruit colour. These lines had a homozygous S. chmielewskii introgression on the short arm of chromosome 1, consistent with the position of the y mutation known to result in colourless epidermis, and hence pink-coloured fruit, when combined with a red flesh. Metabolic analysis showed that pink fruit lack the ripening-dependent accumulation of the yellow-coloured flavonoid naringenin chalcone in the fruit peel, while carotenoid levels are not affected. The expression of all genes encoding biosynthetic enzymes involved in the production of the flavonol rutin from naringenin chalcone were down-regulated in pink fruit, suggesting that the candidate gene underlying the pink phenotype encodes a regulatory protein such as a transcription factor rather than a biosynthetic enzyme. Of 26 MYB and bHLH transcription factors putatively involved in regulating transcription of genes in the phenylpropanoid and/or flavonoid pathway, only the expression level of the MYB12 gene correlated well with the decrease in the expression of structural flavonoid genes in peel samples of pink- and red-fruited genotypes during ripening. Genetic mapping and segregation analysis showed that MYB12 is located on chromosome 1 and segregates perfectly with the characteristic pink fruit colour. Virus-induced gene silencing (VIGS) of SlMYB12 resulted in a decrease in the accumulation of naringenin chalcone, a phenotype consistent with the pink-coloured tomato fruit of IL1b. In conclusion, biochemical and molecular data, gene mapping, segregation analysis and VIGS experiments demonstrated that the MYB12 transcription factor plays an important role in regulating the flavonoid pathway in tomato fruit and suggest strongly that SlMYB12 is a likely candidate for the y mutation.






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