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Anthocyanin accumulation naturally occurs in the ‘blood orange’ (Citrus sinensis (L.) Osbeck) and is controlled by a retrotransposon that is inserted in the promoter of the Ruby transcription factor upon cold stress. Several enzymes work sequentially for anthocyanin biosynthesis following the activation of Ruby. We analyzed the juice vesicles of non-transgenic and the transgenic ‘Mexican’ lime that overexpressed the Ruby transcription factor to understand the regulatory mechanism of anthocyanin biosynthesis in transgenic fruits. Cyanidin and petunidin anthocyanins were detected using liquid chromatography in the juice extracted from Ruby transgenic ‘Mexican’ limes. Additionally, we identified a positive correlation between the overexpression of anthocyanin and the total content of flavonoids and phenolic compounds. In contrast, overexpression of Ruby had no significant effect on the ascorbic acid content or …