Cloning and molecular characterization of the Fatty Acid Elongase 1 (FAE 1) gene from high and low erucic acid lines of Brassica campestris and Brassica oleracea

Das, Sandip; Roscoe, Thomas; Delseny, Michel; Srivastava, Prem S.; Lakshmikumaran, Malathi

doi:10.1016/s0168-9452(01)00556-8

Cited by 48 publications

(24 citation statements)

References 30 publications

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“…The FAE1 polypeptides involved in the synthesis of VLCFAs in Arabidopsis (James et al, 1995) and Brassica seeds (Clemens and Kunst, 1997;Katavic et al, 2002) showed approximately 52% to 54% identity with the nasturtium FAE. The nasturtium FAE protein was predicted to have a theoretical pI value of 9.3 using the algorithm of Bjellqvist et al (1993Bjellqvist et al ( , 1994 and a molecular mass of 56.8 kD, which are similar to the respective values reported for the B. napus CE7 and CE8 FAE homologs (Barret et al, 1998) as well as those from Brassica rapa (campestris) and Brassica oleracea (Das et al, 2002).…”

Section: Isolation Of Nasturtium Fae Homologsupporting

confidence: 73%

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Seed-Specific Heterologous Expression of a Nasturtium FAE Gene in Arabidopsis Results in a Dramatic Increase in the Proportion of Erucic Acid

et al. 2004

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The fatty acid elongase [often designated FAE or b-(or 3-) ketoacyl-CoA synthase] is a condensing enzyme and is the first component of the elongation complex involved in synthesis of erucic acid (22:1) in seeds of garden nasturtium (Tropaeolum majus). Using a degenerate primers approach, a cDNA of a putative embryo FAE was obtained showing high homology to known plant elongases. This cDNA contains a 1,512-bp open reading frame that encodes a protein of 504 amino acids. A genomic clone of the nasturtium FAE was isolated and sequence analyses indicated the absence of introns. Northern hybridization showed the expression of this nasturtium FAE gene to be restricted to the embryo. Southern hybridization revealed the nasturtium b-ketoacyl-CoA synthase to be encoded by a small multigene family. To establish the function of the elongase homolog, the cDNA was introduced into two different heterologous chromosomal backgrounds (Arabidopsis and tobacco [Nicotiana tabacum]) under the control of a seed-specific (napin) promoter and the tandem 35S promoter, respectively. Seed-specific expression resulted in up to an 8-fold increase in erucic acid proportions in Arabidopsis seed oil, while constitutive expression in transgenic tobacco tissue resulted in increased proportions of very long chain saturated fatty acids. These results indicate that the nasturtium FAE gene encodes a condensing enzyme involved in the biosynthesis of very long chain fatty acids, utilizing monounsaturated and saturated acyl substrates. Given its strong and unique preference for elongating 20:1-CoA, the utility of the FAE gene product for directing or engineering increased synthesis of erucic acid is discussed.Very long chain fatty acids (VLCFAs) with 20 carbons or more are widely distributed in nature. In plants, they are mainly found in epicuticular waxes and in the seed oils of a number of plant species, including members of the Brassicaceae, Limnantheceae, Simmondsia, and Tropaeolaceae (Harwood, 1996;Post-Beittenmiller, 1996;Ghanevati and Jaworski, 2001). A strategic goal in oilseed modification is to genetically manipulate high erucic acid germ plasm of the Brassicaceae to increase the content of erucic acid (22:1 D13) and other strategic VLCFAs in the seed oil for industrial niche market needs (Puyaubert et al., 2001;Taylor et al., 2001). Erucic acid and its derivatives are feedstocks in manufacturing slip-promoting agents, surfactants, plasticizers, nylon 1313, and surface coatings; more than 1,000 patents have been issued (Sonntag, 1991(Sonntag, , 1995Leonard, 1994). The current market for high erucate oils exceeds $120 million U.S./annum. Worldwide erucic acid demand is predicted to increase from about 40 million pounds (M pds) in 1990 to about 80 M pds by the year 2010. Similarly, demand for the derivative, behenic acid, is predicted to triple to about 102 M pds by 2010 (Sonntag, 1995). In recent years, production has increased to meet market needs, and high erucic acreage in western Canada is currently at a record high (D. Males, Saskatchewan...

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Section: Isolation Of Nasturtium Fae Homologsupporting

confidence: 73%

“…It also lacks a KXKXX or KKXX motif (X 5 any amino acid) often found at the C terminus of proteins retained within endoplasmic reticulum membranes (Jackson et al, 1990). Rather, it is a type IIIa protein, typically present on endoplasmic reticular membranes (Nakai and Kanehisa, 1992;Das et al, 2002). …”

Section: Isolation Of Nasturtium Fae Homologmentioning

confidence: 99%

Seed-Specific Heterologous Expression of a Nasturtium FAE Gene in Arabidopsis Results in a Dramatic Increase in the Proportion of Erucic Acid

et al. 2004

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“…Closely related orthologs of these genes have been isolated from Brassica species. The B. napus genes FAE1.1 and FAE1.2 have been shown to cosegregate with two loci that govern 22:1n9 production (Barret et al, 1998;Fourmann et al, 1998), with FAE1 genes also being isolated from B. oleracea and B. rapa (Das et al, 2002) as well as B. juncea (Gupta et al, 2004). Between four and six copies of FAD2 have been reported for B. napus (Scheffler et al, 1997) and representative orthologous genes encoding for both FAD2 and FAD3 have been cloned (Arondel et al, 1992;Okuley et al, 1994).…”

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confidence: 99%

Novel Insights into Seed Fatty Acid Synthesis and Modification Pathways from Genetic Diversity and Quantitative Trait Loci Analysis of the Brassica C Genome

et al. 2007

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Natural genetic variation in fatty acid synthesis and modification pathways determine the composition of vegetable oils, which are major components of human diet and renewable products. Based on known pathways we combined diversity and genetic analysis of metabolites to infer the existence of enzymes encoded by distinct loci, and associated these with specific elongation steps or subpathways. A total of 107 lines representing different Brassica genepools revealed considerable variation for 18 seed fatty acid products. The effect of genetic variation within a single biochemical step on subsequent products was demonstrated using a correlation matrix of scatterplots, and by calculating relative step yields. Surprisingly, diploid Brassica oleracea segregating populations had a similar range of variation for individual fatty acids as across the whole genepool. This allowed identification of 22 quantitative trait loci (QTL) associated with activity in the plastid, early stages of synthesis, desaturation, and elongases. Four QTL were assigned to early stages of synthesis, seven to subpathway specific or general elongase activity, one to ketoacyl acyl-carrier protein synthetase, and two each to fatty acid desaturase and either desaturase or fatty acyl-carrier protein thioesterase. An additional 10 QTL had distinct effects but were not assigned specific functions. Where contrasting behavior in more than one subpathway was detected, we inferred QTL specificity for particular combinations of substrate and product. The assignment of enzyme function to QTL was consistent with the known position of some Brassicaeae candidate genes and collinear regions of the Arabidopsis (Arabidopsis thaliana) genome.

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“…Mutations in FAE1 resulted in reduced levels of seed VLCFAs [67,68], and a deficiency in acyl chain elongation activities toward C 18 and C 20 FAs [69]. FAE1 homologs were identified in several plants, Brassica oleracea [70], Brassica campestris [71], Brassica oleacea [71], Simmondsia chinensis [72], Lesquerella fendleri [73] and Limnanthes alba [74] and also in the green algae, Dunaliella salina [75]. In the Arabidopsis genome, 20 predicted FAE1 homologs were found (Fig.…”

Section: Fatty Acid Elongase-3 Ketoacyl Coa Synthases (Fae-3kcss)mentioning

confidence: 99%

Characterization of Enzymes Involved in Fatty Acid Elongation

Paul¹

2007

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Cloning and molecular characterization of the Fatty Acid Elongase 1 (FAE 1) gene from high and low erucic acid lines of Brassica campestris and Brassica oleracea

Cited by 48 publications

References 30 publications

Seed-Specific Heterologous Expression of a Nasturtium FAE Gene in Arabidopsis Results in a Dramatic Increase in the Proportion of Erucic Acid

Seed-Specific Heterologous Expression of a Nasturtium FAE Gene in Arabidopsis Results in a Dramatic Increase in the Proportion of Erucic Acid

Novel Insights into Seed Fatty Acid Synthesis and Modification Pathways from Genetic Diversity and Quantitative Trait Loci Analysis of the Brassica C Genome

Characterization of Enzymes Involved in Fatty Acid Elongation

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