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BIOCHEMICAL PROCESSES AND MACROMOLECULAR STRUCTURES

Seed-Specific Overexpression of an Endogenous Arabidopsis Phytoene Synthase Gene Results in Delayed Germination and Increased Levels of Carotenoids, Chlorophyll, and Abscisic Acid¹

Department of Plant Biology, Swedish University of Agricultural Science, Box 7080, 750 07 Uppsala, Sweden

Phytoene synthase catalyzes the dimerization of two molecules of geranylgeranyl pyrophosphate to phytoene and has been shown to be rate limiting for the synthesis of carotenoids. To elucidate if the capacity to produce phytoene is limiting also in the seed of Arabidopsis (Wassilewskija), a gene coding for an endogenous phytoene synthase was cloned and coupled to a seed-specific promoter, and the effects of the overexpression were examined. The resulting transgenic plants produced darker seeds, and extracts from the seed of five overexpressing plants had a 43-fold average increase of -carotene and a total average amount of -carotene of approximately 260 µg g^–1 fresh weight. Lutein, violaxanthin, and chlorophyll were significantly increased, whereas the levels of zeaxanthin only increased by a factor 1.1. In addition, substantial levels of lycopene and -carotene were produced in the seeds, whereas only trace amounts were found in the control plants. Seeds from the transgenic plants exhibited delayed germination, and the degree of delay was positively correlated with the increased levels of carotenoids. The abscisic acid levels followed the increase of the carotenoids, and plants having the highest carotenoid levels also had the highest abscisic acid content. Addition of gibberellic acid to the growth medium only partly restored germination of the transgenic seeds.

¹ This work was supported by Stiftelsen för Lantbruksforskning, Ulla och Curt Nicolins Stipendiefond, and AstaCaroteneAB.

² These authors contributed equally to this paper.

^* Corresponding author; e-mail kjells{at}mail1.slu.se; fax 46–18–673279.

Received November 3, 2002; returned for revision December 16, 2002; accepted January 26, 2003.

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