caa a22 4500 60620864X CHVBK 20210128101023.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s00425-014-2162-7 doi (NATIONALLICENCE)springer-10.1007/s00425-014-2162-7 Sunflower ( Helianthus annuus ) fatty acid synthase complex: enoyl-[acyl carrier protein]-reductase genes [Elektronische Daten] [Irene González-Thuillier, Mónica Venegas-Calerón, Rafael Garcés, Penny von Wettstein-Knowles, Enrique Martínez-Force] Main conclusion : Enoyl-[acyl carrier protein]-reductases from sunflower. Abstract: A major factor contributing to the amount of fatty acids in plant oils are the first steps of their synthesis. The intraplastidic fatty acid biosynthetic pathway in plants is catalysed by type II fatty acid synthase (FAS). The last step in each elongation cycle is carried out by the enoyl-[ACP]-reductase, which reduces the dehydrated product of β-hydroxyacyl-[ACP] dehydrase using NADPH or NADH. To determine the mechanisms involved in the biosynthesis of fatty acids in sunflower (Helianthus annuus) seeds, two enoyl-[ACP]-reductase genes have been identified and cloned from developing seeds with 75% identity: HaENR1 (GenBank HM021137) and HaENR2 (HM021138). The two genes belong to the ENRA and ENRB families in dicotyledons, respectively. The genetic duplication most likely originated after the separation of di- and monocotyledons. RT-qPCR revealed distinct tissue-specific expression patterns. Highest expression of HaENR1 was in roots, stems and developing cotyledons whereas that of H a ENR2 was in leaves and early stages of seed development. Genomic DNA gel blot analyses suggest that both are single-copy genes. In vivo activity of the ENR enzymes was tested by complementation experiments with the JP1111 fabI(ts) E. coli strain. Both enzymes were functional demonstrating that they interacted with the bacterial FAS components. That different fatty acid profiles resulted infers that the two Helianthus proteins have different structures, substrate specificities and/or reaction rates. The latter possibility was confirmed by in vitro analysis with affinity-purified heterologous-expressed enzymes that reduced the crotonyl-CoA substrate using NADH with different V max. Springer-Verlag Berlin Heidelberg, 2014 Enoyl-[acyl carrier protein]-reductase (ENR) nationallicence Fatty acid synthase (FAS) nationallicence Helianthus nationallicence Oil biosynthesis nationallicence Substrate specificity nationallicence ACP : Acyl carrier protein nationallicence DAF : Days after flowering nationallicence DAG : Days after germination nationallicence ENR : Enoyl-ACP reductase nationallicence EST : Expressed sequence tags nationallicence FAS : Fatty acid synthase nationallicence KAS : Ketoacyl-ACP synthase nationallicence KAR : Ketoacyl-ACP reductase nationallicence González-Thuillier Irene Instituto de la Grasa (CSIC), Av. Padre García Tejero 4, 41012, Seville, Spain aut Venegas-Calerón Mónica Instituto de la Grasa (CSIC), Av. Padre García Tejero 4, 41012, Seville, Spain aut Garcés Rafael Instituto de la Grasa (CSIC), Av. Padre García Tejero 4, 41012, Seville, Spain aut von Wettstein-Knowles Penny Biology Department, Copenhagen University, Ole Maaloees Vej 5, 2200, Copenhagen N, Denmark aut Martínez-Force Enrique Instituto de la Grasa (CSIC), Av. Padre García Tejero 4, 41012, Seville, Spain aut Planta Springer Berlin Heidelberg 241/1(2015-01-01), 43-56 0032-0935 241:1<43 2015 241 425 https://doi.org/10.1007/s00425-014-2162-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00425-014-2162-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- González-Thuillier Irene Instituto de la Grasa (CSIC), Av. Padre García Tejero 4, 41012, Seville, Spain aut NATIONALLICENCE 700 1- Venegas-Calerón Mónica Instituto de la Grasa (CSIC), Av. Padre García Tejero 4, 41012, Seville, Spain aut NATIONALLICENCE 700 1- Garcés Rafael Instituto de la Grasa (CSIC), Av. Padre García Tejero 4, 41012, Seville, Spain aut NATIONALLICENCE 700 1- von Wettstein-Knowles Penny Biology Department, Copenhagen University, Ole Maaloees Vej 5, 2200, Copenhagen N, Denmark aut NATIONALLICENCE 700 1- Martínez-Force Enrique Instituto de la Grasa (CSIC), Av. Padre García Tejero 4, 41012, Seville, Spain aut NATIONALLICENCE 773 0- Planta Springer Berlin Heidelberg 241/1(2015-01-01), 43-56 0032-0935 241:1<43 2015 241 425