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      Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize.

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          Abstract

          Combinatorial nuclear transformation is a novel method for the rapid production of multiplex-transgenic plants, which we have used to dissect and modify a complex metabolic pathway. To demonstrate the principle, we transferred 5 carotenogenic genes controlled by different endosperm-specific promoters into a white maize variety deficient for endosperm carotenoid synthesis. We recovered a diverse population of transgenic plants expressing different enzyme combinations and showing distinct metabolic phenotypes that allowed us to identify and complement rate-limiting steps in the pathway and to demonstrate competition between beta-carotene hydroxylase and bacterial beta-carotene ketolase for substrates in 4 sequential steps of the extended pathway. Importantly, this process allowed us to generate plants with extraordinary levels of beta-carotene and other carotenoids, including complex mixtures of hydroxycarotenoids and ketocarotenoids. Combinatorial transformation is a versatile approach that could be used to modify any metabolic pathway and pathways controlling other biochemical, physiological, or developmental processes.

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          Author and article information

          Journal
          Proc Natl Acad Sci U S A
          Proceedings of the National Academy of Sciences of the United States of America
          Proceedings of the National Academy of Sciences
          1091-6490
          0027-8424
          Nov 25 2008
          : 105
          : 47
          Affiliations
          [1 ] Departament de Producció Vegetal i Ciència Forestal, Universitat de Lleida, Avenue Alcalde Rovira Roure, 191, 25198 Lleida, Spain.
          Article
          0809737105
          10.1073/pnas.0809737105
          2587607
          19011084
          91a0c77b-0832-4a0c-a61e-a474d7bdea37
          History

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