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Identification of Primula “front-end” desaturases with distinct n−6 or n−3 substrate preferences

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Abstract

cDNA clones encoding cytochrome b5 fusion desaturases were isolated from Primula cortusoides L. and Primula luteola Ruprecht, species previously shown to preferentially accumulate either n−6 or n−3 Δ6-desaturated fatty acids, respectively. Functional characterisation of these desaturases in yeast revealed that the recombinant Primula enzymes displayed substrate preferences, resulting in the predominant synthesis of either γ-linolenic acid (n−6) or stearidonic acid (n−3). Independent expression of the two Primula desaturases in transgenic Arabidopsis thaliana confirmed these results, with γ-linolenic acid and stearidonic acid accumulating in both leaf and seed tissues to different levels, depending on the substrate specificity of the desaturase. Targeted lipid analysis of transgenic Arabidopsis lines revealed the presence of Δ6-desaturated fatty acids in the acyl-CoA pools of leaf but not seed tissue. The implications for the transgenic synthesis of C20 polyunsaturated fatty acids via the elongation of Δ6-desaturated fatty acids are discussed, as is the potential of using Primula desaturases in the synthesis of C18 n−3 polyunsaturated fatty acids such as stearidonic acid.

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Abbreviations

ALA:

α-Linolenic acid

ARA:

Arachidonic acid

DAG:

Diacylglycerol

EPA:

Eicosapentaenoic acid

GLA:

γ-Linolenic acid

LA:

Linoleic acid

LC-PUFA:

Long chain polyunsaturated fatty acid

MGDG:

Monogalactosyl diacylglycerol

ORF:

Open reading frame

PC:

Phosphatidylcholine

SDA:

Stearidonic acid

TAG:

Triacylglycerol

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Acknowledgements

Rothamsted Research receives grant-aided support from BBSRC (UK). The authors gratefully acknowledge the support of BASF Plant Sciences.

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Authors and Affiliations

  1. Rothamsted Research, Harpenden, Herts, AL5 2JQ, UK

    Olga Sayanova, Richard Haslam, Mónica Venegas-Calerón & Johnathan A. Napier

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  1. Olga Sayanova
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  2. Richard Haslam
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  3. Mónica Venegas-Calerón
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  4. Johnathan A. Napier
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Correspondence to Johnathan A. Napier.

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Sayanova, O., Haslam, R., Venegas-Calerón, M. et al. Identification of Primula “front-end” desaturases with distinct n−6 or n−3 substrate preferences. Planta 224, 1269–1277 (2006). https://doi.org/10.1007/s00425-006-0306-0

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