Abstract
The fatty acid elongase 1 (FAE1) gene is a key gene in the erucic acid biosynthesis in rapeseed. The complete coding sequences of the FAE1 gene were isolated separately from eight high and zero erucic acid rapeseed cultivars (Brassica napus L.). A four base pair deletion between T1366 and G1369 in the FAE1 gene was found in a number of the cultivars, which leads to a frameshift mutation and a premature stop of the translation after the 466th amino acid residue. This deletion was predominantly found in the C-genome and rarely in the A-genome of B. napus. Expression of the gene isoforms with the four base pair deletion in a yeast system generated truncated proteins with no enzymatic activity and could not produce very long chain fatty acids as the control with an intact FAE1 gene did in yeast cells. In the developing rape seeds the FAE1 gene isoforms with the four base pair deletion were transcribed normally but failed to translate proteins to form a functional complex. The four base pair deletion proved to be a mutation responsible for the low erucic acid trait in rapeseed and independent from the point mutation reported by Han et al. (Plant Mol Biol 46:229–239, 2001).
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Aherne F, Bowland J, Christian R, Hardin R (1976) Performance of myocardial and blood seral changes in pigs fed diets containing high or low erucic acid rapeseed oils. Can J Anim Sci 56:275–284
Anand I, Downey R (1981) A study of erucic acid alleles in digenomic rapeseed (Brassica napus L.). Can J Plant Sci 61:199–203
Badawy I, Atta B, Ahmed W (1994) Biochemical and toxicological studies on the effect of high and low erucic acid rapeseed oil on rats. Nahrung 38:402–411
Barret P, Delourme R, Renard M, Domergue F, Lessire R, Delseny M, Roscoe T (1998) A rapeseed FAE1 gene is linked to the E1 locus associated with variation in the content of erucic acid. Theor Appl Genet 96:177–186
Beare-Rogers J, Nera E, Heggtveit H (1971) Cardiac lipid changes in rats fed oils containing long–chain fatty acids. Can Inst Food Technol 4:120–124
Blacklock B, Jaworski J (2002) Studies into factors contributing to substrate specificity of membrane-bound 3-ketoacyl-CoA synthases. Eur J Biochem 269:4789–4798
Davies C, Heath RJ, White SW, Rock C (2000) The 1.8 A crystal structure and active-site architecture of beta-ketoacyl-acyl carrier protein synthase III (FabH) from Escherichia coli. Structure 8:185–195
Downey R, Craig B (1964) Genetic control of fatty acid biosynthesis in rapeseed (Brassica napus L.). J Am Oil Chem 41:475–478
Ferrer J, Jez J, Bowman M, Dixon R, Noel J (1999) Structure of chalcone synthase and the molecular basis of plant polyketide biosynthesis. Nat Struct Biol 6:775–784
Fourmann M, Barret P, Renard M, Pelletier G, Delourme R, Brunel D (1998) The two genes homologous to Arabidopsis FAE1 co-segregate with the two loci governing erucic acid content in Brassica napus. Theor Appl Genet 96:852–858
Ghanevati M, Jaworski J (2001) Active-site residues of a plant membrane-bound fatty acid elongase β-ketoacyl-CoA synthase, FAE1 KCS. Biochim Biophys Acta 1530:77–85
Ghanevati M, Jan G (2002) Engineering and mechanistic studies of the Arabidopsis FAE1 β-ketoacyl-CoA synthase, FAE1 KCS. Eur J Biochem 269:3531–3539
Han J, Lühs W, Sonntag K, Zähringer U, Borchardt D, Wolter1 F, Heinz1 E, Frentzen1 M (2001) Functional characterization of β-ketoacyl-CoA synthase genes from Brassica napus L. Plant Mol Biol 46:229–239
Harvey B, Downey R (1963) The inheritance of erucic acid content in rapeseed (Brassica napus L). Can J Plant Sci 44:104–111
Huang W, Jia J, Edwards P, Dehesh1 K, Schneider G, Lindqvist Y (1998) Crystal structure of β-ketoacyl-acyl carrier protein synthase II from E. coli reveals the molecular architecture of condensing enzymes. EMBO J 17:1183–1191
James DW, Lim E, Keller J, Plooy I, Ralston E, Dooner H (1995) Directed tagging of the Arabidopsis FATTY ACID ELONGATION 1 (FAE1) gene with the maize transposon activator. Plant Cell 7:301–319
Jez J, Ferrer J, Bowman M, Dixon R, Noel J (2000) Dissection of malonyl-CoA decarboxylation from polyketide formation in the reaction mechanism of a plant polyketide synthase. Biochemistry 39:890–902
Jönsson R (1977) Erucic-acid heredity in rapeseed (Brassica napus L. and Brassica campestris L.). Hereditas 86:159–170
Katavic V, Barton D, Giblin E, Reed D, Kumar A, Taylor D (2004) Gaining insight into the role of serine 282 in B. napus FAE1 condensing enzyme. FEBS Lett 562:118–124
Katavic V, Mietkiewska E, Barton D, Giblin E, Reed D, Taylor D (2002) Restoring enzyme activity in nonfunctional low erucic acid Brassica napus fatty acid elongase 1 by a single amino acid substitution. Eur J Biochem 269:5625–5631
Laemmli U (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Lassner M, Lardizabal K, Metz J (1996) A jojoba β-ketoacyl-CoA synthase cDNA complements the canola fatty acid elongation mutation in transgenic plants. Plant Cell 8:281–292
Lemieux B, Miquel M, Somerville C, Browse J (1990) Mutants of Arabidopsis with alterations in seed lipid fatty acid composition. Theor Appl Genet 80:234–240
Liu H (1985) Rapeseed genetics and breeding. Shanghai Scientific and Technology Publishing House, Shanghai (in Chinese)
Olsen J, Kadziola A, von Wettstein-Knowles P, Siggaard-Andersen M, Lindquist Y, Larsen S (1999) The X-ray crystal structure of beta-ketoacyl [acyl carrier protein] synthase I. FEBS Lett 460:46–52
Puyaubert J, Garbay B, Costaglioli P, Dieryck W, Roscoe T, Renard M, Cassagne C, Lessire R (2001) Acyl-CoA elongase expression during seed development in Brassica napus. Biochim Biophys Acta 1533:141–152
Qiu X, Janson C, Konstantinidis A, Nwagwu S, Silverman C, Smith W, Khandekar S, Lonsdale J, Abdel-Meguid S (1999) Crystal structure of beta-ketoacyl-acyl carrier protein synthase III. A key condensing enzyme in bacterial fatty acid biosynthesis. J Biol Chem 274:36465–36471
Roscoe T, Lessire R, Puyaubert J, Renard M, Delseny M (2001) Mutations in the fatty acid elongation 1 gene are associated with a loss of β-ketoacyl-CoA synthase activity in low erucic acid rapeseed. FEBS Lett 492:107–111
Saghai-Maroof MA, Soliman K, Jorgensen R, Allard R (1984) Ribosomal DNA spacer-length polymorphisms in barley: mendelian inheritance, chromosomal location, and population dynamics. Proc Natl Acad Sci USA 81:8014–8018
Sambrook J, Fritsch E, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New York
Todd J, Post-Beittenmiller D, Jaworski J (1999) KCS1 encodes a fatty acid elongase 3-ketoacyl-CoA synthase affecting wax biosynthesis in Arabidopsis thaliana. Plant J 17:119–130
Wu Y, Xiao L, Wu G, Lu C (2007) Cloning of fatty acid elongase1 gene and molecular identification of A and C genome in Brassica species. Sci China Ser C-Life Sci 3:343–349
Acknowledgments
This work was financially supported by the Development Plan of the State Key Fundamental Research of China (2006CB101600), the National Foundation of Natural Sciences of China (30471099) and the National High Technology and Development Program of China (2006AA10A113).
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Communicated by D. A. Lightfoot.
Gang Wu, Yuhua Wu contribute equally to this article.
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Wu, G., Wu, Y., Xiao, L. et al. Zero erucic acid trait of rapeseed (Brassica napus L.) results from a deletion of four base pairs in the fatty acid elongase 1 gene. Theor Appl Genet 116, 491–499 (2008). https://doi.org/10.1007/s00122-007-0685-z
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DOI: https://doi.org/10.1007/s00122-007-0685-z