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Plant Physiol, February 2002, Vol. 128, pp. 439-453
Functional Analysis of the Early Steps of Carotenoid Biosynthesis
in Tobacco
Marco
Busch,
Anja
Seuter, and
Rüdiger
Hain*
Bayer AG, Agricultural Division Research, Molecular Target Research
and Biotechnology, 51368 Leverkusen, Germany
Carotenoids contribute to energy transduction in the light
harvesting complexes and serve in protection from excess light fluence.
Because of the importance of carotenoids, the genes encoding enzymes of
carotenoid biosynthesis in higher plants are potential targets for
herbicides. To obtain further insight into tobacco carotenoid
biosynthesis and to investigate and prioritize potential herbicide
targets in the pathway, the effects of changed phytoene synthase (PSY)
and phytoene desaturase (PDS) gene expression were studied in
transgenic tobacco (Nicotiana tabacum Petit Havana SR1)
plants. Genes for both enzymes were cloned from tobacco, and
surprisingly two functional PSY genes were found. Transgenic tobacco
plants constitutively expressing these genes in both sense and
antisense orientations were examined regarding phenotype, carotenoid
content and transcript levels of carotene biosynthesis genes.
Overexpression of either psy gene resulted in severe
phenotypic effects including dwarfism, altered leaf morphology, and
pigmentation. A correlation among phenotype, transcript level, and
metabolic profile was demonstrated by comparison of hemizygous and
homozygous plants from the same transformation event. Antisense
expression of PSY and PDS also caused lethal phenotypes. Transcript
levels of other carotene biosynthesis genes remained unaltered in the transgenic mutant. Phytoene accumulated in plants expressing antisense RNA to pds. However, elevated levels of phytoene were
detected suggesting an increase in metabolic flux into this pathway.
*
Corresponding author; e-mail ruediger.hain.rh{at}bayer-ag.de;
fax 0049-0-2173-38-3520.
© 2002 American Society of Plant Physiologists
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