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Molecular Cloning and Expression Analysis of Genes Encoding Two Microsomal Oleate Desaturases (FAD2) from Safflower (Carthamus tinctorius L.)

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Abstract

The endoplasmic reticulum (ER)-associated oleate desaturase (FAD2) is the key enzyme responsible for the production of linoleic acid in plants. In safflower, a seed-type FAD2 gene (CtFAD2-1) has been isolated previously. In this study, two different cDNA sequences, designated CtFAD2-2 and CtFAD2-3, encoding two microsomal oleate desaturases have been isolated from safflower (Carthamus tinctorius L.) using a polymerase chain reaction (PCR) approach. Both deduced amino acid sequences showed the three histidine boxes characteristic of all membrane-bound desaturases, and possess a C-terminal ER retention signal. Phylogentic analysis shows that CtFAD2-2 is grouped with other house-keeping type FAD2 sequences from oil crops, while CtFAD2-3 does not belong to seed type or house-keeping type FAD2. Real-time PCR analysis showed that CtFAD2-2 gene is constitutively expressed in both vegetative tissues and developing seeds, with higher transcript levels in roots, stems and petioles. Transcript of CtFAD2-3 was strongly detected in developing seeds, showing low levels in vegetative tissues. In response to low-temperature stress, CtFAD2-2 and CtFAD2-3 in different tissues showed a different behavior (up- or down-regulated). The transcript of CtFAD2-2 was dramatically decreased in roots, stems and petioles under low temperature, while CtFAD2-3 showed a significant increase in these tissues. On the contrary, in leaves, the strongly enhanced expression of CtFAD2-2 was observed under low temperature, while the transcript of CtFAD2-3 showed a little decrease.

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Authors and Affiliations

  1. Agronomy College, Sichuan Agricultural University, Ya’an, 625014, China

    Ling-Liang Guan, Yuan-Biao Wang, Hao Shen, Kai Hou, Ying-Wen Xu & Wei Wu

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  1. Ling-Liang Guan
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  2. Yuan-Biao Wang
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  3. Hao Shen
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  4. Kai Hou
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  5. Ying-Wen Xu
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  6. Wei Wu
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Correspondence to Wei Wu.

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Guan, LL., Wang, YB., Shen, H. et al. Molecular Cloning and Expression Analysis of Genes Encoding Two Microsomal Oleate Desaturases (FAD2) from Safflower (Carthamus tinctorius L.). Plant Mol Biol Rep 30, 139–148 (2012). https://doi.org/10.1007/s11105-011-0322-5

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