Skip to main content
SpringerLink
Log in

Increased levels of glycerol-3-phosphate lead to a stimulation of flux into triacylglycerol synthesis after supplying glycerol to developing seeds of Brassica napus L. in planta

  • Original Article
  • Published:
Planta Aims and scope Submit manuscript

Abstract

Glycerol-3-phosphate (glycerol-3P) is a primary substrate for triacylglycerol synthesis. In the present study, changes in the levels of glycerol-3P during rape (Brassica napus L.) seed development and the influence of manipulating glycerol-3P levels on triacylglycerol synthesis were investigated. (i) Glycerol-3P levels were high in young seeds and decreased during seed development at 30 and 40 days after flowering (DAF), when lipid accumulation was maximal. (ii) To manipulate glycerol-3P levels in planta, various concentrations of glycerol were injected directly into 30-DAF seeds, which remained otherwise intact within their siliques and attached to the plant. Injection of 0–10 nmol glycerol led to a progressive increase in seed glycerol-3P levels within 28 h. (iii). Increased levels of glycerol-3P were accompanied by an increase in the flux of injected [14C]sucrose into total lipids and triacylglycerol, whereas fluxes to organic acids, amino acids, starch, protein and cell walls were not affected. (iv) When [14C]acetate was injected into seeds, label incorporation into total lipids and triacylglycerol increased progressively with increasing glycerol-3P levels. (v) There was a strong correlation between the level of glycerol-3P and the incorporation of injected [14C]acetate and [14C]sucrose into triacylglycerol. (v) The results provide evidence that the prevailing levels of glycerol-3P co-limit triacylglycerol synthesis in developing rape seeds.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1a, b
Fig. 2a, b
Fig. 3a, b
Fig. 4a–j
Fig. 5a–h
Fig. 6a–f
Fig. 7a, b

Similar content being viewed by others

Abbreviations

DAF:

Days after flowering

DAG:

Diacylglycerol

G3PAT:

Glycerol-3-phosphate acyltransferase

Glycerol-3P:

Glycerol-3-phosphate

PA:

Phosphatidic acid

PC:

Phosphatidylcholine

TAG:

Triacylglycerol,

References

  • Bafor M, Jonsson L, Stobart AK, Stymne S (1990) Regulation of triacylglycerol biosynthesis in embryos and microsomal preparations from the developing seeds of Cuphea lanceolata. Biochem J 272:31–38

    CAS  PubMed  Google Scholar 

  • Bao XM, Ohlrogge J (1999) Supply of fatty acid is one limiting factor in the accumulation of triacylglycerol in developing embryos. Plant Physiol 120:1057–1062

    Article  CAS  Google Scholar 

  • Bao XM, Focke M, Pollard M, Ohlrogge JB (2000) Understanding in vivo carbon precursor supply for fatty acid synthesis in leaf tissue. Plant J 22:39–50

    PubMed  Google Scholar 

  • Benning C, Sommerville CR (1992) Identification of an operon involved in sulfolipid biosynthesis in Rhodobacter sphaeroides. J Bacteriol 174:6479–6487

    CAS  Google Scholar 

  • Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Phys 37:911–917

    CAS  Google Scholar 

  • Brough CL, Coventry JM, Christie WW, Kroon AP, Brown AP, Barsby TL, Slabas AR (1996). Towards the genetic engineering of triacylglycerols of defined fatty acid composition: major changes in erucic acid content at the sn-2 position affected by the introduction of a 1-acyl-sn-glycerol-3-phosphate acyltransferases from Limnanthes douglasii into oil seed rape. Mol Breed 2:133–142

    CAS  Google Scholar 

  • Cao YZ, Huang AHC (1986) Diacylglycerol acyltransferases in maturing oilseeds of maize and others species. Plant Physiol 82:813–820

    CAS  Google Scholar 

  • Eastmond PJ, Rawsthorne S (2000) Coordinate changes in carbon partitioning and plastidial metabolism during the development of oilseed rape embryos. Plant Physiol 122:767–774

    Article  CAS  Google Scholar 

  • Eccleston VS, Harwood JL (1995) Solubilization partial-purification and properties of acyl-coA-glycerol-3-phosphate acyltransferases from avocado (Persea americana) fruit mesocarp. BBA-Lipid Lipid Met 1257:1–10

    Google Scholar 

  • Finlayson SA, Dennis DT (1980) NAD+-specific glycerol 3-phosphate dehydrogenase from developing castor bean endosperm. Arch Biochem Biophys 199:179–185

    CAS  Google Scholar 

  • Gardiner SE, Heinz E, Roughan PG (1984) Rates and products of long-chain fatty-acid synthesis from [1-C-14]-labeled acetate in chloroplasts isolated from leaves of 16-3 and 18-3 plants. Plant Physiol 74:890–896

    CAS  Google Scholar 

  • Gee R, Goyala A, Gerber D, Byerrum RU, Tolbert NE (1988) Isolation of dihydroxyacetone phosphate reductase from Dunaliella chloroplasts and comparison with isozymes from spinach leaves. Plant Physiol 88:896–903

    CAS  Google Scholar 

  • Geigenberger P, Reimholz R, Geiger M, Merlo L, Canale V, Stitt M (1997) Regulation of sucrose and starch metabolism in potato tubers in response to short-term water deficit. Planta 201:502–518

    Article  CAS  Google Scholar 

  • Gibon Y, Vigeolas H, Tiessen A, Geigenberger P, Stitt M (2002) Sensitive and high throughput metabolite assays for inorganic pyrophosphate, ADPGlc, nucleotide phosphates, and glycolytic intermediates based on a novel enzymic cycling system. Plant J 30:221–235

    Article  CAS  Google Scholar 

  • Heldt HW (1997) Plant biochemistry and molecular biology. Oxford University Press, New York

  • Hippmann H, Heinz E (1976) Glycerol kinase in leaves. Z Pflanzenphysiol 79:408–418

    CAS  Google Scholar 

  • Huang AHC (1975a) Enzymes of glycerol metabolism in storage tissues of fatty seedlings. Plant Physiol 55:555–558

    CAS  Google Scholar 

  • Huang AHC (1975b) Enzymes of glycerol metabolism in storage tissues of fatty seedlings. Plant Physiol 56:84–84

    Google Scholar 

  • Jako J, Kumar A, Wie Y, Zou J, Barton DL, Giblin EM, Covello PS, Taylor DC (2001) Seed-specific over-expression of an Arabidopsis cDNA encoding a diacylglycerol acyltransferase enhances seed oil content and seed weight. Plant Physiol 126:861–874

    Article  CAS  PubMed  Google Scholar 

  • Katavic V, Reed DW, Taylor DC, Giblin, Barton DL, Zou JT, Mackenzie SL, Covello PS, Kunst L (1995) Alteration of seed fatty-acid composition by an ethyl methanesulfonate-induced mutation in Arabidopsis thaliana affecting diacylglycerol acyltransferase activity. Plant Physiol 108:399–409

    Article  CAS  PubMed  Google Scholar 

  • Kirsch T, Gerber DW, Byerrum RU, Tolbert NE (1992) Plant dihydroxyacetone phosphate reductases—purification, characterization, and localization. Plant Physiol 100:352–359

    CAS  Google Scholar 

  • Kennedy EP (1961) Biosynthesis of complex lipids. Fed Proc Fed Am Soc Exp Biol 20:934

    CAS  Google Scholar 

  • Kubis SE, Rawsthorne S (2000) The role of plastidial transporters in developing embryos of oilseed rape (Brassica napus L.) for fatty acid synthesis. Biochem Soc Trans 28:665–666

    Article  CAS  Google Scholar 

  • Lacey DJ, Hills MJ (1996) Heterogeneity of the endoplasmic reticulum with respect to lipid synthesis in developing seeds of Brassica napus L. Planta 199:545–551

    CAS  Google Scholar 

  • Lacey DJ, Wellner N, Beaudoin F, Napier JA, Shewry PR (1998) Secondary structure of oleosins in oil bodies isolated from seeds of safflower (Carthamus tinctorius L.) and sunflower (Helianthus annuus L.). Biochem J 334:469–477

    CAS  PubMed  Google Scholar 

  • Lassner MW, Levering CK, Davies HM, Knutzen DS (1995) Lysophosphatidic acid acyltransferases from Meadowfoam mediates insertion of erucic-acid at the sn-2 position of triacylglycerol in transgenic rapeseed oil. Plant Physiol 109:1389–1394

    Article  CAS  Google Scholar 

  • Manaf AM, Harwood JL (2000) Purification and characterization of acyl-coA: glycerol 3-phosphate acyltransferases from oil palm (Elaeis guineensis) tissues. Planta 210:318–328

    CAS  PubMed  Google Scholar 

  • Merlo L, Geigenberger P, Hajirezaei M, Stitt M (1993) Changes of carbohydrates, metabolites and enzyme activities in potato tubers during development, and within a single tuber along a stolon-apex gradient. J Plant Physiol 142:392–402

    CAS  Google Scholar 

  • Murphy DJ (1994) Biogenesis, function and biotechnology of plant-storage lipids. Prog Lipid Res 33:71–85

    Article  CAS  PubMed  Google Scholar 

  • Ohlrogge J, Browse J (1995) Lipid biosynthesis. Plant Cell 7:957–970

    CAS  PubMed  Google Scholar 

  • Ohlrogge JB, Jaworski JG (1997) Regulation of fatty acid synthesis. Annu Rev Plant Physiol Plant Mol Biol. 48:109–136

    Google Scholar 

  • Perry HJ, Harwood JL (1993) Changes in the lipid content of developing rape seeds of Brassica napus. Phytochemistry 32:1411–1415

    Article  CAS  Google Scholar 

  • Perry HJ, Bligny R, Gout E, Harwood JL (1999) Changes in Kennedy pathway intermediates associated with increased triacylglycerol synthesis in oil-seed rape. Phytochemistry 52:799–804

    Article  CAS  Google Scholar 

  • Sadava D, Moore K (1987) Glycerol metabolism in higher plants—glycerol kinase. Biochem Biophys Res Commun 143:977–983

    CAS  PubMed  Google Scholar 

  • Sellwood C, Slabas AR, Rawsthorne S (2000) Effects of manipulating expression of acetylcoA carboxylase in Brassica napus L. embryos. Biochem Soc Trans 28:598–600

    Article  CAS  Google Scholar 

  • Sharma N, Phutela A, Malhotra SP, Singh R (2001) Purification and characterization of dihydroxyacetone phosphate reductase from immature seeds of Brassica campestris L. Plant Sci 160:603–610

    Article  CAS  Google Scholar 

  • Slack CR, Roughan PG, Balasingham N (1978) Labelling of glycerolipids in the cotyledons of developing oilseeds by [1-14C] acetate and [2-3H] glycerol. Biochem J 170:421–433

    CAS  Google Scholar 

  • Stobart K, Mancha M, Lenman M, Dahlqvist A, Stymne S (1997) Triacylglycerols are synthesised and utilized by transacylation reactions in microsomal preparations of developing safflower (Carthamus tinctorius L) seeds. Planta 203:58–66

    Article  CAS  Google Scholar 

  • Stymne S, Stobart AK (1987) Triacylglycerol biosynthesis. In: Stumpf PK (ed) The biochemistry of plants, vol 9. Academic Press, New York, pp 175–214

  • Zou J, Katavic V, Giblin EM, Barton DL, Mackenzie SL, Keller WA, HU X, Taylor DC (1997) Modification of seed oil content and acyl composition in the Brassicaceae by expression of a yeast sn-2 acyltransferase gene. Plant Cell 9:909–923

    Google Scholar 

Download references

Acknowledgements

We thank Mark Stitt for his support and interest in this work and helpful comments on the manuscript. We are grateful to Peter Dörmann for providing the GC facilities, to Peter Dörmann and John Lunn for critical readings of the manuscript, to Karin Koehl and Britta Hausmann for taking care of the plants, and to Peter Waldeck for technical help (all MPI Molecular Plant Physiology, Golm, Germany). This work was supported by BASF Plant Science GmbH (Ludwigshafen, Germany).

Author information

Authors and Affiliations

  1. Max Planck Institute of Molecular Plant Physiology, Am Mühlenberg 1, 14476, Golm, Germany

    Helene Vigeolas & Peter Geigenberger

Authors
  1. Helene Vigeolas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peter Geigenberger
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Peter Geigenberger.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vigeolas, H., Geigenberger, P. Increased levels of glycerol-3-phosphate lead to a stimulation of flux into triacylglycerol synthesis after supplying glycerol to developing seeds of Brassica napus L. in planta . Planta 219, 827–835 (2004). https://doi.org/10.1007/s00425-004-1273-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00425-004-1273-y

Keywords

Search

Navigation