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Metabolism of glucose-6-phosphate and utilization of multiple metabolites for fatty acid synthesis by plastids from developing oilseed rape embryos

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Abstract

The aim of this work was to investigate the partitioning of imported glucose 6-phosphate (Glc6P) to starch and fatty acids, and to CO2 via the oxidative pentose phosphate pathway (OPPP) in plastids isolated from developing embryos of oilseed rape (Brassica napus L.). The ability of the isolated plastids to utilize concurrently supplied substrates and the effects of these substrate combinations on the Glc6P partitioning were also assessed. The relative fluxes of carbon from Glc6P to starch, fatty acids, and to CO2 via the OPPP were close to 2∶1∶1 when Glc6P was supplied alone. Under these conditions NADPH generated via the OPPP was greater than that required by the concurrent rate of fatty acid synthesis. Fatty acid synthesis was unaffected by the presence or absence of exogenous NADH and/or NADPH and the requirement of fatty acid synthesis for reducing power is therefore met entirely by intraplastidial metabolism. When Glc6P was supplied in the presence of either pyruvate or pyruvate and acetate, the total flux from these metabolites to fatty acids was up to threefold greater than that from either Glc6P or pyruvate when they were supplied singly. In these experiments there was little competition between Glc6P and pyruvate in fatty acid synthesis and the flux to starch was unchanged. This implies that the starch and fatty acid biosynthesis pathways did not compete for the exogenously supplied ATP on which they were strongly dependent. When Glc6P and pyruvate were provided together, the NADPH generated by the OPPP pathway was less than that required by the concurrent rate of fatty acid synthesis. This suggests that the metabolism of exogenous Glc6P via the OPPP can contribute to the NADPH demand created during fatty acid synthesis but it also indicates that other intraplastidial sources of reducing power must be available under the in-vitro conditions used.

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Abbreviations

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

Glc6P:

glucose-6-phosphate

OPPP:

oxidative pentose phosphate pathway

PIM:

plastid isolation medium

Pyr:

pyruvate

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Additional information

The authors are grateful to Drs A.M. Smith and K. Denyer for helpful and stimulating discussions throughout this study. We thank Drs D.J. Murphy and M.J. Hills for comments on the manuscript. Mrs S. Fincham is also thanked for glasshouse assistance. Seed of B. napus cv. Topas was kindly provided by Dalgety Agriculture (Essex, UK). This work was supported by the Biotechnology and Biological Sciences Research Council through grant-in-aid to the John Innes Centre and by a Cambridge Laboratory studentship to Fan Kang.

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Kang, F., Rawsthorne, S. Metabolism of glucose-6-phosphate and utilization of multiple metabolites for fatty acid synthesis by plastids from developing oilseed rape embryos. Planta 199, 321–327 (1996). https://doi.org/10.1007/BF00196575

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Key words

  • Brassica
  • Embryo
  • Carbon partitioning
  • Fatty acid synthesis
  • Plastid
  • Starch synthesis