Advertisement

Planta

, Volume 235, Issue 3, pp 629–639 | Cite as

Reduced expression of FatA thioesterases in Arabidopsis affects the oil content and fatty acid composition of the seeds

  • Antonio J. Moreno-Pérez
  • Mónica Venegas-Calerón
  • Fabián E. Vaistij
  • Joaquín J. Salas
  • Tony R. Larson
  • Rafael Garcés
  • Ian A. Graham
  • Enrique Martínez-Force
Original Article

Abstract

Acyl–acyl carrier protein (ACP) thioesterases are enzymes that control the termination of intraplastidial fatty acid synthesis by hydrolyzing the acyl–ACP complexes. Among the different thioesterase gene families found in plants, the FatA-type fulfills a fundamental role in the export of the C18 fatty acid moieties that will be used to synthesize most plant glycerolipids. A reverse genomic approach has been used to study the FatA thioesterase in seed oil accumulation by screening different mutant collections of Arabidopsis thaliana for FatA knockouts. Two mutants were identified with T-DNA insertions in the promoter region of each of the two copies of FatA present in the Arabidopsis genome, from which a double FatA Arabidopsis mutant was made. The expression of both forms of FatA thioesterases was reduced in this double mutant (fata1 fata2), as was FatA activity. This decrease did not cause any evident morphological changes in the mutant plants, although the partial reduction of this activity affected the oil content and fatty acid composition of the Arabidopsis seeds. Thus, dry mutant seeds had less triacylglycerol content, while other neutral lipids like diacylglycerols were not affected. Furthermore, the metabolic flow of the different glycerolipid species into seed oil in the developing seeds was reduced at different stages of seed formation in the fata1 fata2 line. This diminished metabolic flow induced increases in the proportion of linolenic and erucic fatty acids in the seed oil, in a similar way as previously reported for the wri1 Arabidopsis mutant that accumulates oil poorly. The similarities between these two mutants and the origin of their phenotype are discussed in function of the results.

Keywords

Acyl–acyl carrier protein thioesterase Deficient mutant FatA Metabolic flux Oil content Triacylglycerols 

Abbreviations

TE

Acyl–ACP thioesterase

KASII

β-Ketoacyl-ACP synthase II

SAD

Stearoyl-ACP desaturase

ODS

Oleate desaturase

LDS

Linoleate desaturase

FAE

Fatty acid elongase

Notes

Acknowledgments

We are grateful to Rosario Sánchez and Valeria Gazda for their technical assistance. We also thank Dr. Luisa Hernández for help with experimental approach. This work was supported by the Spanish MICINN and FEDER, Project AGL2008-01086/ALI.

References

  1. Andre C, Froehlich JE, Moll MR, Benning C (2007) A heteromeric plastidic pyruvate kinase complex involved in seed oil biosynthesis. Plant Cell 19:2006–2022PubMedCrossRefGoogle Scholar
  2. Baud S, Wuillème S, Dubreucq B, de Almeida A, Vuagnat C, Lepiniec L, Miquel M, Rochat C (2007) Function of plastidial pyruvate kinase in seeds of Arabidopsis thaliana. Plant J 52:405–419PubMedCrossRefGoogle Scholar
  3. Beisson F, Koo AJK, Ruuska S, Schwender J, Pollard M, Thelen JJ, Paddock T, Salas JJ, Savage L, Milcamps A, Mhaske VB, Cho Y, Ohlrogge JB (2003) Arabidopsis genes involved in acyl lipid metabolism. A 2003 census of the candidates, a study of the distribution of expressed sequence tags in organs, and a Web-based database. Plant Physiol 132:681–697PubMedCrossRefGoogle Scholar
  4. Bonaventure G, Salas JJ, Pollard MR, Ohlrogge JB (2003) Disruption of the FATB gene in Arabidopsis demonstrates an essential role of saturated fatty acids in plant growth. Plant Cell 15:1020–1033PubMedCrossRefGoogle Scholar
  5. Bonaventure G, Bao X, Ohlrogge J, Pollard M (2004) Metabolic responses to the reduction in palmitate caused by disruption of the FATB gene in Arabidopsis. Plant Physiol 135:1269–1279PubMedCrossRefGoogle Scholar
  6. Browse J, Somerville CR (1991) Glycerolipid synthesis: biochemistry and regulation. Annu Rev Plant Physiol Plant Mol Biol 42:467–506CrossRefGoogle Scholar
  7. Burgal J, Shockey J, Lu C, Dyer J, Larson T, Graham I, Browse J (2008) Metabolic engineering of hydroxy fatty acid production in plants: RcDGAT2 drives dramatic increases in ricinoleate levels in seed oil. Plant Biotechnol J 8:819–831CrossRefGoogle Scholar
  8. Cernac A, Benning C (2004) WRINKLED1 encodes an AP2/EREB domain protein involved in the control of storage compound biosynthesis in Arabidopsis. Plant J 40:575–585PubMedCrossRefGoogle Scholar
  9. Chen M, Han G, Dietrich CR, Dunn TM, Cahoon EB (2006) The essential nature of sphingolipids in plants as revealed by the functional identification and characterization of the Arabidopsis LCB1 subunit of serine palmitoyltransferase. Plant Cell 18:3576–3593PubMedCrossRefGoogle Scholar
  10. Dehesh K, Jones A, Knutzon DS, Voelker TA (1996) Production of high levels of 8:0 and 10:0 fatty acids in transgenic canola by overexpression of Ch FatB2, a thioesterase cDNA from Cuphea hookeriana. Plant J 9:167–172PubMedCrossRefGoogle Scholar
  11. Dörmann P, Kridl JC, Ohlrogge JB (1994) Cloning and expression in Escherichia coli of a cDNA coding for the oleoyl-acyl carrier protein thioesterase from coriander (Coriandrum sativum L.). Biochim Biophys Acta Lipids Lipid Metab 1212:134–136CrossRefGoogle Scholar
  12. Dörmann P, Voelker TA, Ohlrogge JB (2000) Accumulation of palmitate in Arabidopsis mediated by the acyl-acyl carrier protein thioesterase FATB1. Plant Physiol 123:637–644PubMedCrossRefGoogle Scholar
  13. Focks N, Benning C (1998) Wrinkled1: a novel, low-seed-oil mutant of Arabidopsis with a deficiency in the seed-specific regulation of carbohydrate metabolism. Plant Physiol 118:91–101PubMedCrossRefGoogle Scholar
  14. Hellyer A, Leadlay PF, Slabas AR (1992) Induction, purification and characterisation of acyl-ACP thioesterase from developing seeds of oil seed rape (Brassica napus). Plant Mol Biol 20:1573–5028CrossRefGoogle Scholar
  15. Jiang P, Cronan JE (1994) Inhibition of fatty-acid synthesis in Escherichia coli in the absence of phospholipid-synthesis and release of inhibition by thioesterase action. J Bacteriol 176:2814–2821PubMedGoogle Scholar
  16. Jones A, Davies HM, Voelker TA (1995) Palmitoyl-acyl carrier protein (ACP) thioesterase and the evolutidnary-origin of plant acyl-ACP thioesterases. Plant Cell 7:359–371PubMedCrossRefGoogle Scholar
  17. Joyard J, Stumpf PK (1980) Characterization of an acyl-coenzyme A thioesterase associated with the envelope of spinach chloroplasts. Plant Physiol 65:1039–1043PubMedCrossRefGoogle Scholar
  18. Koo AJK, Ohlrogge JB, Pollard M (2004) On the export of fatty acids from the chloroplast. J Biol Chem 279:16101–16110PubMedCrossRefGoogle Scholar
  19. Larson TR, Graham IA (2001) A novel technique for the sensitive quantification of acyl CoA esters from plant tissues. Plant J 25:115–125PubMedCrossRefGoogle Scholar
  20. Larson TR, Edgell T, Byrne J, Dehesh K, Graham IA (2002) Acyl CoA profiles of transgenic plants that accumulate medium-chain fatty acids indicate inefficient storage lipid synthesis in developing oilseeds. Plant J 32:519–527PubMedCrossRefGoogle Scholar
  21. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408PubMedCrossRefGoogle Scholar
  22. Lu YJ, Zhang YM, Rock CO (2004) Product diversity and regulation of type II fatty acid synthases. Biochem Cell Biol 82:145–155PubMedCrossRefGoogle Scholar
  23. Maeo K, Tokuda T, Ayame A, Mitsui N, Kawai T, Tsukagoshi H, Ishiguro S, Nakamura K (2009) An AP2-type transcription factor, WRINKLED1, of Arabidopsis thaliana binds to the AW-box sequence conserved among proximal upstream regions of genes involved in fatty acid synthesis. Plant J 60:476–487PubMedCrossRefGoogle Scholar
  24. Martínez-Force E, Cantisán S, Serrano-Vega MJ, Garcés R (2000) Acyl–acyl carrier protein thioesterase activity from sunflower (Helianthus annuus L.) seeds. Planta 211:673–678PubMedCrossRefGoogle Scholar
  25. Mayer KM, Shanklin J (2005) A structural model of the plant acyl-acyl carrier protein thioesterase FatB comprises two helix/4-stranded sheet domains, the N-terminal domain containing residues that affect specificity and the C-terminal domain containing catalytic residues. J Biol Chem 280:3621–3627PubMedCrossRefGoogle Scholar
  26. McLaren I, Wood C, Jalil MNH, Yong BCS, Thomas DR (1985) Carnitine acyltransferases in chloroplasts of Pisum sativum L. Planta 163:197–200CrossRefGoogle Scholar
  27. Ohlrogge JB, Jaworski JG (1997) Regulation of fatty acid synthesis. Annu Rev of Plant Physiol Plant Mol Biol 48:109–136CrossRefGoogle Scholar
  28. Pollard M, Ohlrogge J (1999) Testing models of fatty acid transfer and lipid synthesis in spinach leaf using in vivo oxygen-18 labeling. Plant Physiol 121:1217–1226PubMedCrossRefGoogle Scholar
  29. Rock CO, Garwin JL (1979) Preparative enzymatic synthesis and hydrophobic chromatography of acyl-acyl carrier protein. J Biol Chem 254:7123–7128PubMedGoogle Scholar
  30. Roughan PG, Slack CR (1982) Cellular organization of glycerolipid metabolism. Annu Rev Plant Physiol 33:97–132CrossRefGoogle Scholar
  31. Ruuska SA, Girke T, Benning C, Ohlrogge JB (2002) Contrapuntal networks of gene expression during Arabidopsis seed filling. Plant Cell 14:1191–1206PubMedCrossRefGoogle Scholar
  32. Salas JJ, Ohlrogge JB (2002) Characterization of substrate specificity of plant FatA and FatB acyl-ACP thioesterases. Arch Biochem Biophys 403:25–34PubMedCrossRefGoogle Scholar
  33. Sánchez-García A, Moreno-Pérez AJ, Muro-Pastor AM, Salas JJ, Garcés R, Martínez-Force E (2010) Acyl–ACP thioesterases from castor (Ricinus communis L.): an enzymatic system appropriate for high rates of oil synthesis and accumulation. Phytochemistry 71:860–869PubMedCrossRefGoogle Scholar
  34. Schmid M, Davison TS, Henz SR, Pape UJ, Demar M, Vingron M, Schölkopf B, Weigel D, Lohmann JU (2005) A gene expression map of Arabidopsis thaliana development. Nat Genet 37:501–506PubMedCrossRefGoogle Scholar
  35. Serrano-Vega MJ, Garcés R, Martínez-Force E (2005) Cloning, characterization and structural model of a FatA-type thioesterase from sunflower seeds (Helianthus annuus L.). Planta 221:868–880PubMedCrossRefGoogle Scholar
  36. Thelen JJ, Ohlrogge JB (2002) Both antisense and sense expression of biotin carboxyl carrier protein isoform 2 inactivates the plastid acetyl-coenzyme A carboxylase in Arabidopsis thaliana. Plant J 32:419–431PubMedCrossRefGoogle Scholar
  37. Thomas DR, Jalil MNH, Ariffin A, Cooke RJ, McLaren I, Yong BCS, Wood C (1983) The synthesis of short- and long-chain acylcarnitine by etioplasts of greening barley leaves. Planta 158:259–263CrossRefGoogle Scholar
  38. Voelker TA, Worrell AC, Anderson L, Bleibaum J, Fan C, Hawkins DJ, Radke SE, Davies HM (1992) Fatty acid biosynthesis redirected to medium chains in transgenic oilseed plants. Science 257:72–74PubMedCrossRefGoogle Scholar
  39. Voelker TA, Jones A, Cranmer AM, Davies HM, Knutzon DS (1997) Broad-range and binary-range acyl–acyl-carrier-protein thioesterases suggest an alternative mechanism for medium-chain production in seeds. Plant Physiol 114:669–677PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Antonio J. Moreno-Pérez
    • 1
  • Mónica Venegas-Calerón
    • 1
  • Fabián E. Vaistij
    • 2
  • Joaquín J. Salas
    • 1
  • Tony R. Larson
    • 2
  • Rafael Garcés
    • 1
  • Ian A. Graham
    • 2
  • Enrique Martínez-Force
    • 1
  1. 1.Instituto de la GrasaSevilleSpain
  2. 2.Centre for Novel Agricultural Products, Department of BiologyUniversity of YorkYorkUK

Personalised recommendations