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Acta Agron Sin ›› 2013, Vol. 39 ›› Issue (05): 928-934.doi: 10.3724/SP.J.1006.2013.00928

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Functional Characterization of Isochrysis galbana Δ5 Desaturase Gene IgD5 in Arabidopsis thaliana

LIU Jiang**,SUN Quan-Xi**,LI Xin-Zheng*,QI Bao-Xiu*   

  1. State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Tai’an 271018, China
  • Received:2012-09-07 Revised:2013-01-15 Online:2013-05-12 Published:2013-02-19
  • Contact: 亓宝秀, E-mail: qbx126@sdau.edu.cn, Tel: 0538-8246205; 李新征, E-mail: lxz@sdau.edu.cn, Tel: 0538-8246205

Abstract:

We have isolated a Δ5 desaturase gene from the alga Isochrysis galbana, namely IgD5. Phylogenetic analysis showed that it shared the highest homology with a functionally characterized Δ5 desaturase from Pavlova salina. Previously, we demonstrated that IgD5 has activity towards the omega-6 fatty acid substrate by heterologous expression of IgD5 inSaccharomyces cerevisiae. However, whether IgD5 also has activity towards the potential omega-3 fatty acid substrate has not been verified. In this study, we transformed IgD5 into a double transgenic Arabidopsis thaliana (TMA2)that can already synthesize di-homo-γ-linolenic acid (DGLA, 20:3Δ8,11,14) and eicosatetraenoic acid (ETA, 20:4Δ8,11,14,17). Triple transgenic plants were identified by PCR and RT-PCR. The total fatty acids composition of the transgenic lines was determined by gas chromatography. Novel fatty acids, arachidonic acid (AA, 20:4Δ5, 8,11,14) and eicosapentaenoic acid (EPA, 20:5Δ5,8,11,14,17) were detected and accounted for 7.44% and 2.07% of total fatty acids, with a conversion rate of 68% and 60%, respectively, which are much higher than those in yeast. In addition, IgD5 did not show a preference for either ω3 or ω6 substrate. Furthermore, IgD5 has strong substrate specificity as no fatty acid products were detected except for AA and EPA. These new findings further confirmed that IgD5 is an excellent gene for the production of fish oil in plants due to its high enzyme activity coupled with substrate specificity.

Key words: Δ5 desaturase, Arachidonic acid, Eicosapentaenoic acid, ω3 fatty acids, Arabidopsis thaliana

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