Molecular Analysis of Carotenoid Biosynthesis in Plants: Characterizing the Genes Psy, Pds and CrtL-e

In this research we have studied the molecular biology of carotenoid biosynthesis in tomato. The investigations focused on the genes Pds and Psy, encoding desaturase and phytoene synthase, respectively, which are key enzymes in the biosynthetic pathway of lycopene and b-carotene. In addition, we have investigated the genes for lycopene cyclase. We have cloned from tomato and characterized the cDNA of CrtL-e, which encodes the lycopene e-cyclase, and analyzed its expression during fruit development. The results establish a paradigm for the regulation of carotenoid pigment biosynthesis during the ripening process of fruits. It is concluded that transcriptional regulation of genes that encode carotenoid-biosynthesis enzymes is the major mechanism that governs specific pigment accumulation. During the ripening of tomato fruits transcription of the genes encoding the enzymes phytoene synthase and phytoene desaturase is up-regulated, while the transcription of the genes for both lycopene cyclases decreases and thus the conversion of lycopene to subsequent carotenoids is inhibited. These findings support the working hypothesis of the molecular approach to manipulating carotenogenesis by altering gene expression in transgenic plants, and offer obvious strategies to future application in agriculture. The molecular and physiological knowledge on carotenogenesis gained in this project, suggest a concept for manipulating gene expression that will alter carotenoid composition in fruits and flowers.