caa a22 4500 467894396 CHVBK 20180406152758.0 cr unu---uuuuu 170328e20060601xx s 000 0 eng 10.1007/s11103-006-0012-0 doi (NATIONALLICENCE)springer-10.1007/s11103-006-0012-0 Cloning, Functional Identification and Sequence Analysis of Flavonoid 3′-hydroxylase and Flavonoid 3′,5′-hydroxylase cDNAs Reveals Independent Evolution of Flavonoid 3′,5′-hydroxylase in the Asteraceae Family [Elektronische Daten] [Christian Seitz, Christian Eder, Bettina Deiml, Sandra Kellner, Stefan Martens, Gert Forkmann] Flavonoids are ubiquitous secondary plant metabolites which function as protectants against UV light and pathogens and are involved in the attraction of pollinators as well as seed and fruit dispersers. The hydroxylation pattern of the B-ring of flavonoids is determined by the activity of two members of the vast and versatile cytochrome P450 protein (P450) family, the flavonoid 3′-hydroxylase (F3′H) and flavonoid 3′,5′-hydroxylase (F3′5′H). Phylogenetic analysis of known sequences of F3′H and F3′5′H indicated that F3′5′H was recruited from F3′H before the divergence of angiosperms and gymnosperms. Seven cDNAs were isolated from species of the Asteraceae family, all of which were predicted to code for F3′Hs based on their sequences. The recombinant proteins of four of the heterologously in yeast expressed cDNAs exhibited the expected F3′H activity but surprisingly, three recombinant proteins showed F3′5′H activity. Phylogenetic analyses indicated the independent evolution of an Asteraceae-specific F3′5′H. Furthermore, sequence analysis of these unusual F3′5′H cDNAs revealed an elevated rate of nonsynonymous substitutions as typically found for duplicated genes acquiring new functions. Since F3′5′H is necessary for the synthesis of 3′,4′,5′-hydroxylated delphinidin-derivatives, which normally provide the basis for purple to blue flower colours, the evolution of an Asteraceae-specific F3′5′H probably reflects the adaptive value of efficient attraction of insect pollinators. Springer, 2006 Asteraceae nationallicence cytochrome P450 nationallicence evolution of flavonoid 3′-hydroxylase and flavonoid 3′,5′-hydroxylase nationallicence flavonoid biosynthesis nationallicence gene duplication nationallicence P450 : cytochrome P450 protein nationallicence DHK : dihydrokaempferol nationallicence DHM : dihydromyricetin nationallicence DHQ : dihydroquercetin nationallicence ERI : eriodictyol nationallicence F3′H : flavonoid 3′-hydroxylase nationallicence F3′5′H : flavonoid 3′,5′-hydroxylase nationallicence Ka : rate of nonsynonymous substitutions per nonsynonymous site nationallicence Ks : rate synonymous substitutions per synonymous site nationallicence NAR : naringenin nationallicence PHF : pentahydroxyflavanone nationallicence TLC : thin layer chromatography nationallicence Seitz Christian Chair of Floriculture Crops and Horticultural Plant Breeding, Technical University Munich, Am Hochanger 4, 85350, Freising, Germany aut Eder Christian Competence Network Malignant Lymphomas, University Cologne, 50924, Cologne, Germany aut Deiml Bettina Chair of Floriculture Crops and Horticultural Plant Breeding, Technical University Munich, Am Hochanger 4, 85350, Freising, Germany aut Kellner Sandra Institute of Pharmaceutical Biology, Philipps-University Marburg, D-35037, Marburg/Lahn, Germany aut Martens Stefan Institute of Pharmaceutical Biology, Philipps-University Marburg, D-35037, Marburg/Lahn, Germany aut Forkmann Gert Chair of Floriculture Crops and Horticultural Plant Breeding, Technical University Munich, Am Hochanger 4, 85350, Freising, Germany aut Plant Molecular Biology Kluwer Academic Publishers 61/3(2006-06-01), 365-381 0167-4412 61:3<365 2006 61 11103 https://doi.org/10.1007/s11103-006-0012-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11103-006-0012-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Seitz Christian Chair of Floriculture Crops and Horticultural Plant Breeding, Technical University Munich, Am Hochanger 4, 85350, Freising, Germany aut NATIONALLICENCE 700 1- Eder Christian Competence Network Malignant Lymphomas, University Cologne, 50924, Cologne, Germany aut NATIONALLICENCE 700 1- Deiml Bettina Chair of Floriculture Crops and Horticultural Plant Breeding, Technical University Munich, Am Hochanger 4, 85350, Freising, Germany aut NATIONALLICENCE 700 1- Kellner Sandra Institute of Pharmaceutical Biology, Philipps-University Marburg, D-35037, Marburg/Lahn, Germany aut NATIONALLICENCE 700 1- Martens Stefan Institute of Pharmaceutical Biology, Philipps-University Marburg, D-35037, Marburg/Lahn, Germany aut NATIONALLICENCE 700 1- Forkmann Gert Chair of Floriculture Crops and Horticultural Plant Breeding, Technical University Munich, Am Hochanger 4, 85350, Freising, Germany aut NATIONALLICENCE 773 0- Plant Molecular Biology Kluwer Academic Publishers 61/3(2006-06-01), 365-381 0167-4412 61:3<365 2006 61 11103 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer