Cloning and Functional Analysis of Fatty Desaturase Gene (FAD2-1) Promoter from Safflower (Carthamus tinctorious)
LI Dan-Dan1,2, WANG Qing1, XU Xin1, YU Jing-Sheng1, WU Wei1,*
1 College of Agriculture, Sichuan Agricultural University, Chengdu 611130, China; 2 Key Laboratory of Medicinal Plant Breeding and Planting in Guizhou, Guiyang 550025, China
Abstract：Linoleic acid is main polyunsaturated fatty acids in plant lipids, which plays key roles in plant metabolism. Fatty desaturase 2 (FAD2) is a key enzyme for linoleic acid biosynthesis, CtFAD2-1 gene has the highest activity and was mainly responsible for the biosynthesis of linoleic acid in safflower (Carthamus tinctorius) seed. In this study, an 821 bp promoter region of CtFAD2-1 gene from safflower was cloned. Four 5'-deletion constructs of 821 bp (P1), 564 bp (P2), 230 bp (P3), 60 bp (P4) were constructed and functionally characterized in Arabidopsis thaliana by using transgenic approach. Computational analysis found that the abiotic stress responsive cis-elements like heat responsive element (HSE), low temperature response (LTR), MYB response (MBS) etc, as well as wound, fungus, methyljasmonate responsive motifs existed in the promoter. The histochemical staining of GUS revealed that the flower, mature seed and silique wall were stained. In addition, the transgenic plants treated by drought, salt, cold, high-temperature, wound and ABA hormone as well as control plants were also stained. The -230~-60 bp might be the key region for GUS activity in A. thaliana tissues. CtFAD2-1 gene was not seed-specific and it could be induced by an assortment of abiotic stresses. This research plays an indispensable role in furthering the research related to the regulation of CtFAD2-1 gene.
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