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Sunflower (Helianthus annuus) fatty acid synthase complex: enoyl-[acyl carrier protein]-reductase genes

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Abstract

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.

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Abbreviations

ACP:

Acyl carrier protein

DAF:

Days after flowering

DAG:

Days after germination

ENR:

Enoyl-ACP reductase

EST:

Expressed sequence tags

FAS:

Fatty acid synthase

KAS:

Ketoacyl-ACP synthase

KAR:

Ketoacyl-ACP reductase

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Acknowledgments

We thank Dr. Alicia M. Muro-Pastor for help with Southern blot analysis and A. González-Callejas and B. Lopez-Cordero for skilful technical assistance. This work was supported by the "Ministerio de Economia y Competitividad" and FEDER, project AGL2011-23187. IGT was supported by a JAE-CSIC contract in part financed by European Social Fund.

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Authors and Affiliations

  1. Instituto de la Grasa (CSIC), Av. Padre García Tejero 4, 41012, Seville, Spain

    Irene González-Thuillier, Mónica Venegas-Calerón, Rafael Garcés & Enrique Martínez-Force

  2. Biology Department, Copenhagen University, Ole Maaloees Vej 5, 2200, Copenhagen N, Denmark

    Penny von Wettstein-Knowles

Authors
  1. Irene González-Thuillier
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  2. Mónica Venegas-Calerón
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  3. Rafael Garcés
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  4. Penny von Wettstein-Knowles
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  5. Enrique Martínez-Force
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Correspondence to Mónica Venegas-Calerón.

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González-Thuillier, I., Venegas-Calerón, M., Garcés, R. et al. Sunflower (Helianthus annuus) fatty acid synthase complex: enoyl-[acyl carrier protein]-reductase genes. Planta 241, 43–56 (2015). https://doi.org/10.1007/s00425-014-2162-7

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