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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 |
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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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Received: 13 July 2020
Published: 01 April 2021
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Corresponding Authors:
* ewuwei@gmail.com
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[1] 官玲亮. 2011. 红花(Carthamus tinctorius L.)不同组织多不饱和脂肪酸积累模式及调控机制[D]. 博士学位论文, 四川农业大学, 导师: 吴卫, pp.98-130. (Guan L L.2011. Accumulation pattern and regulatory mechanisms of fatty acid in different safflower (Carthamus tinctorius L.) tissues[D]. Thesis for Ph.D., Sichuan Agriculture University, Supervisor: Wu W, pp. 98-130.) [2] 李丹丹, 王庆, 胡博, 等. 2018. 低温对红花脂肪酸组成和脂肪酸脱氢酶基因表达的影响[J]. 分子植物育种, 16(16): 5223-5231. (Li D D, Wang Q,Hu B, et al.2018, Effects of low temprature on fatty acid composition and gene expression of fatty acid desaturase in safflower[J]. Molecular plant breeding, 16(16): 5223-5231.) [3] 任超翔, 吴沂芸, 唐小慧. 等. 2017. 红花的起源和产地变迁[J]. 中国中药杂志, 42(11): 2219-2222. (Ren C X, W Y Y, Tang X H, et al.2017. Safflower's origin and changes of producing areas[J]. China Journal of Chinese Materia Medica, 42(11): 2219-2222.) [4] 孙亮. 2018. 棉花FAD2-1基因的5'UTR内含子与启动子的顺式作用元件的鉴定及其转录调控作用的研究[D]. 硕士学位论文, 石河子大学, 导师: 孙杰, 31-44. (Sun L.2018. The cis-element identification and transcription regulation of the promoter and 5'UTR of FAD2-1 gene in cotton[D]. Thesis for Ph.D., Shihezi University, Supervisor: Sun J, 31-44.) [5] Bruxelles G L D, Peacock W J, Dennis E S, et al.1996. Abscisic acid induces the alcohol dehydrogenase gene in Arabidopsis[J]. Plant Physiology, 111(2): 381-391. [6] Cao H, Wang L, Nawaz M A, et al.2017. Ectopic expression of pumpkin NAC transcription factor CmNAC1 improves multiple abiotic stress tolerance in Arabidopsis[J]. Frontiers in Plant Science, 8: 2052-2065. [7] Cao S, Zhou X R, Wood C C, et al.2013. A large and functionally diverse family of FAD2 genes in safflower (Carthamus tinctorius L.)[J]. BMC Plant Biology, 13: 5. [8] Dar A A, Choudhury A R, Kancharla P K, et al.2017. The FAD2 gene in plants: Occurrence, regulation, and role[J]. Frontiers in Plant Science, 8: 1789-1804. [9] Guan, L L, Xu Y W, Wang Y B, et al.2012. Isolation and characterization of a temperature-regulated microsomal oleate desaturase gene (CtFAD2-1) from safflower (Carthamus tinctorius L.)[J]. Plant Molecular Biology Reporter, 30(2): 391-402. [10] Guiltinan M J, Marcotte W R, Quatrano R S.1990. A plant leucine zipper protein that recognizes an abscisic acid response element[J]. Science, 250(4978): 267-271. [11] Kim M J, Kim H, Shin J S, et al.2006. Seed-specific expression of sesame microsomal oleic acid desaturase is controlled by combinatorial properties between negative cis-regulatory elements in the SeFAD2 promoter and enhancers in the 5'-UTR intron[J]. Molecular and Genetics Genomics, 276(4): 351-368. [12] Kim M J, Kim J K, Shin J S, et al.2007. The SebHLH transcription factor mediates trans-activation of the SeFAD2 gene promoter through binding to E- and G-box elements[J]. Plant Molecular Biology, 64(4): 453-466. [13] Li D D, Yu J S, Wang Q, et al.2019. Sequence variations and expression analysis of FAD2 among different LA-type safflower materials[J]. Acta Physiologiae Plantarum, 41: 135-140. [14] Magali L, Patrice D, Gert T, et al.2002. PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences[J]. Nucleic Acids Research, 30(1): 325-327. [15] Mártínez-Rivas J M, Sperling P, Lȕhs W, et al.2001. Heinz, E. Spatial and temporal regulation of three differentmicrosomal oleate desaturase genes (FAD2) from normal-type and high-oleic varieties of sunflower (Helianthus anmuus L.)[J]. Molecular Breeding, 8: 159-168 [16] Narusaka Y, Nakashima K, Shinwari ZK, et al.2003. Interaction between two cis-acting elements, ABRE and DRE, in ABA-dependent expression of Arabidopsis rd29A gene in response to dehydration and high-salinity stresses[J]. The Plant Journal, 34(2): 137-148. [17] Niu G L, Gou W, Han X L, et al.2018. Cloning and functional analysis of phosphoethanolamine methyltransferase promoter from maize (Zea mays L.)[J]. International Journal of Moleceular Sciences, 19(1): 191-203. [18] Peng X, Li, H, Wang D, et al.2016. Genome-wide identification of the Jatropha curcas, MYB family and functional analysis of the abiotic stress responsive gene JcMYB2[J]. BMC Genomics, 17(1): 251-256. [19] Rathod R P.2018. Cost and returns of safflower in marathwada region of maharashtra[J]. Periodic Research, 3(6): 28-29. [20] Roy S.2015. Function of MYB domain transcription factors in abiotic stress and epigenetic control of stress response in plant genome[J]. Plant Signaling & Behavior, 11(1): e1117723. [21] Tao Y, Luo L, He L, et al.2014. A promoter analysis of Mother of ft and tfl11 (JcMFT1), a seed-preferential gene from the biofuel plant Jatropha curcas[J]. Journal of Plant Research, 127(4): 513-524. [22] Weber H.2002. Fatty acid-derived signals in plants[J]. Trends in Plant Science, 7: 217-224. [23] Zhang J, Liu H, Sun, J, et al.2012. Arabidopsis fatty acid desaturase FAD2 is required for salt tolerance during seed germination and early seedling growth[J]. PLOS ONE, 7(1): e30355. |
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