Genome-wide identification and expression profiling of duplicated flavonoid 3'-hydroxylase gene family in Carthamus tinctorius L.
Flavonoid 3′-hydroxylase (F3’H) enzyme is essential in determining the flavonoids B-ring hydroxylation pattern. It is mainly implicated in the biosynthetic pathway of cyaniding-based anthocyanins, flavonols, and flavan-3-ols. However, the evolution and regulatory mechanism of these important flavonoid hydroxylases have not been systematically investigated in safflower (Carthamus tinctorius L.). In this study, we identified 22 duplicatedCtF3'H-encoding genes from safflower through genome-wide prediction and conservation analysis. Phylogenetic analysis revealed the pattern of conservation and divergence of CtF3'Hs encoding proteins and their homologs from different plant species. The distribution of conserved protein motifs and cis-regulatory units suggested several structural components that could be crucial in deciphering the final function of CtF3'H proteins. Furthermore, the results of RNA-seq and qRT-PCR assay in different flowering tissues suggested differential expression level of CtF3’H genes during flower development. Based on the unique homology of CtF3’H5 with flavonoid 3’ hydroxylases from other plant species, further validation of CtF3’H5 was carried out. The transient expression of CtF3’H5 in onion epidermal cells implied that the subcellular localization of the fusion construct containing CtF3’H5 and GFP was predominantly detected in the plasma membrane. Similarly, the prokaryotic expression and western blot hybridization of CtF3’H5 demonstrated the detection of a stable 50.3kD target protein. However, more efforts are needed to further extend the functional validation of CtF3’H5 in safflower. This study provides a fundamental gateway for future functional studies and understanding the genetic evolution of F3'Hs in plants.
Ahmad N, Jianyu L, Xu T, Noman M, Jameel A, Na Y, … Fawei W (2019). Overexpression of a novel cytochrome P450 promotes flavonoid biosynthesis and osmotic stress tolerance in transgenic Arabidopsis. Genes 10(10):756. https://doi.org/10.3390/genes10100756
Ahmad N, Li T, Liu Y, Hoang NQV, Ma X, Zhang X, Liu J, Yao N, Liu X, Li H (2020). Molecular and biochemical rhythms in dihydroflavonol 4-reductase-mediated regulation of leucoanthocyanidin biosynthesis in Carthamus tinctorius L. Industrial Crops and Products 156:112838. https://doi.org/10.1016/j.indcrop.2020.112838
Alaiye A, Kaya E, Pnarbal MZ, Harmanc N, Cingi C (2020). An experimental comparison of the analgesic and anti-inflammatory effects of Safflower oil, benzydamine HCl, and naproxen sodium. Journal of Medicinal Food 23(8). https://doi.org/10.1089/jmf.2019.0157
Ambreen H, Kumar S, Kumar A, Agarwal M, Jagannath A, Goel S (2018). Carthamus tinctorius association mapping for important agronomic traits in safflower (L.) Core collection using microsatellite markers. Frontiers in plant science 9:402. https://doi.org/10.3389/fpls.2018.00402
Barvkar VT, Pardeshi VC, Kale SM, Kadoo NY, Gupta VS (2012). Phylogenomic analysis of UDP glycosyltransferase 1 multigene family in Linum usitatissimum identified genes with varied expression patterns. BMC Genomics 13(1):175-175. https://doi.org/10.1186/1471-2164-13-175
Biasini M, Bienert S, Waterhouse A, Arnold K, Studer G, Schmidt T, … Bordoli L (2014). SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information. Nucleic Acids Research 42(W1):W252-W258. https://doi.org/10.1093/nar/gku340
Castellarin SD, Di Gaspero G, Marconi R, Nonis A, Peterlunger E, Paillard S, … Testolin R (2006). Colour variation in red grapevines (Vitis vinifera L.): genomic organisation, expression of flavonoid 3'-hydroxylase, flavonoid 3'5'-hydroxylase genes and related metabolite profiling of red cyanidin-/blue delphinidin-based anthocyanins in berry skin. BMC Genomics 7(1):1-17. https://doi.org/10.1186/1471-2164-7-12
Cerqueira A, Santana I, Araújo J, Lima H, Batatinha M, Branco A, Santos Junior M, Botura M (2021). Homology modeling, docking, molecular dynamics and in vitro studies to identify Rhipicephalus microplus acetylcholinesterase inhibitors. Journal of Biomolecular Structure & Dynamics 1-11. https://doi.org/10.1080/07391102.2021.1889666
Chen K, Liu H, Lou Q, Liu Y (2017). Ectopic expression of the grape hyacinth R2R3-MYB transcription factor gene, induces anthocyanin accumulation in tobacco. Frontiers in Plant Science 8:965. https://doi.org/10.3389/fpls.2017.00965
Cho H, Kim O, Lee Y, Kang L, Nguyen C, Ishihara A, Kim H (2017). Feruloylserotonin inhibits hydrogen peroxide-induced melanogenesis and apoptosis in B16F10 and SK-Mel-2 melanoma cells. Biochemical and Biophysical Research Communications 491(4):973-979. https://doi.org/10.1016/j.bbrc.2017.07.158
Choudhary D, Pandey A, Adhikary S, Ahmad N, Trivedi R (2016). Genetically engineered flavonol enriched tomato fruit modulates chondrogenesis to increase bone length in growing animals. Scientific Reports 6:21668. https://doi.org/10.1038/srep21668
De Palma M, Fratianni F, Nazzaro F, Tucci M (2014). Isolation and functional characterization of a novel gene coding for flavonoid 3’-hydroxylase from globe artichoke. Biologia Plantarum 58(3):445-455. https://doi.org/10.1007/s10535-014-0424-7
Dermauw W, Van Leeuwen T, Feyereisen R (2020). Diversity and evolution of the P450 family in arthropods. Insect Biochemistry and Molecular Biology 127:103490. https://doi.org/10.1016/j.ibmb.2020.103490.
Du C, Hou J, Wang C, Zhang M, Zheng Y, Yang G, Hu Y (2021). Effects of safflower yellow on cholesterol levels in serum and brain tissue of APP/PS1 mice. Metabolic Brain Disease 36(4):557-569. https://doi.org/10.1007/s11011-021-00680-0
Gao, S, Xu X, Zeng W, Xu S, Lyv Y, Feng Y, … Chen J (2020). Efficient Biosynthesis of (2)-Eriodictyol from (2)-Naringenin in through a combination of promoter adjustment and directed evolution. ACS Synthetic Biology 9(12):3288-3297. https://doi.org/10.1021/acssynbio.0c00346
Guo L, Gao L, Ma X, Guo F, Ruan H, Bao Y, … Wang Y (2019). Functional analysis of flavonoid 3'-hydroxylase and flavonoid 3',5'-hydroxylases from tea plant (Camellia sinensis), involved in the B-ring hydroxylation of flavonoids. Gene 717:144046. https://doi.org/10.1016/j.gene.2019.144046
Han Y, Vimolmangkang S, Soria-Guerra RE, Rosales-Mendoza S, Zheng D, Lygin AV, Korban SS (2010). Ectopic expression of apple F3’H genes contributes to anthocyanin accumulation in the Arabidopsis tt7 mutant grown under nitrogen stress. Plant Physiology 153(2):806-820. https://doi.org/10.1104/pp.109.152801
Hong Y, Ahmad N, Tian Y, Liu J, Wang L, Wang G, … Liu W (2019). Genome-wide identification, expression analysis, and subcellular localization of Carthamus tinctorius bHLH transcription factors. International Journal of Molecular Sciences 20(12):3044. https://doi.org/10.3390/ijms20123044.
Iwashina T, Benitez ER, Takahashi R (2006) Analysis of flavonoids in pubescence of soybean near-isogenic lines for pubescence color loci. Journal of Heredity 97:438-443. https://doi.org/10.1093/jhered/esl027
Jia Y, Li B, Zhang Y, Zhang X, Xu Y, Li C (2019). Evolutionary dynamic analyses on monocot flavonoid 3’-hydroxylase gene family reveal evidence of plant-environment interaction. BMC Plant Biology 19(1):1-16. https://doi.org/10.1186/s12870-019-1947-z
Knowles P (1972). The plant geneticist’s contribution toward changing lipid and amino acid composition of safflower. Journal of the American Oil Chemists Society 49(1):30-32. https://doi.org/10.1007/BF02545134
Li H, Li L, Shang Guan G, Jia C, Deng S, Noman M, … Zhang X (2020). Genome-wide identification and expression analysis of bZIP gene family in Carthamus tinctorius L. Scientific Reports 10(1):1-15. https://doi.org/10.1038/s41598-020-72390-z
Lin K, Qin Z, Qu C, Chen X, Jiang Q, Li M, … Li D (2021). Hydroxyl safflower yellow B combined with doxorubicin inhibits the proliferation of human breast cancer MCF-7 cells. Oncology Letters 21(5):426. https://doi.org/10.3892/ol.2021.12687
Liu X, Ahmad N, Yang L, Fu T, Kong J, Yao N, Dong Y, Wang N, Li X, Wang F (2019). Molecular cloning and functional characterization of chalcone isomerase from Carthamus tinctorius. AMB Express 9(1):1-12. https://doi.org/10.1186/s13568-019-0854-x
Lu R, Yuan T, Wang Y, Zhang T, Yuan Y, Wu D, … Cheng Y (2018). Spontaneous severe hypercholesterolemia and atherosclerosis lesions in rabbits with deficiency of low-density lipoprotein receptor (LDLR) on exon 7. EBioMedicine 36:29-38. https://doi.org/10.1016/j.ebiom.2018.09.020
Lv GY, Guo XG, Xie LP, Xie CG, Zhang XH, Yuan Y, … Guo AG (2017). Molecular characterization, gene evolution, and expression analysis of the fructose-1, 6-bisphosphate aldolase (FBA) gene family in wheat (Triticum aestivum L.). Frontiers in Plant Science 8. https://doi.org/10.3389/fpls.2017.01030.
Min F, Sun H, Wang B, Ahmad N, Li H (2020). Hepatoprotective effects of hydroxysafflor yellow A in D-galactose-treated aging mice. European Journal of Pharmacology 173214. https://doi.org/10.1016/j.ejphar.2020.173214
Mizuno H, Yazawa T, Kasuga S, Sawada Y, Kanamori H, Ogo Y, Hirai MY, Matsumoto T, Kawahigashi H (2016). Expression of flavone synthase II and flavonoid 3’-hydroxylase is associated with color variation in tan-colored injured leaves of sorghum. Frontiers in Plant Science 7:1718. https://doi.org/10.3389/fpls.2016.01718
Musiol-Kroll EM, Tocchetti A, Sosio M, Stegmann E (2019). Challenges and advances in genetic manipulation of filamentous actinomycetes – the remarkable producers of specialized metabolites. Natural Product Reports 36(12). https://doi.org/10.1039/C9NP00029A
Nagamatsu A, Masuta C, Matsuura H, Kitamura K, Abe J, Kanazawa A (2009). Down-regulation of flavonoid 3’-hydroxylase gene expression by virus-induced gene silencing in soybean reveals the presence of a threshold mRNA level associated with pigmentation in pubescence. Journal of Plant Physiology 166(1):0-39. https://doi.org/10.1016/j.jplph.2008.02.002
Naqvi AAT, Anjum F, Khan FI, Islam A, Hassan MI (2016). Sequence analysis of hypothetical proteins from Helicobacter pylori 26695 to identify potential virulence factors. Genomics Informatics 14(3). https://dx.doi.org/10.5808%2FGI.2016.14.3.125
Nelson D, Werck‐Reichhart D (2011). A P450‐centric view of plant evolution. The Plant Journal 66(1):194-211. https://doi.org/10.1111/j.1365-313X.2011.04529.x
Panche A, Diwan A, Chandra S (2016). Flavonoids: an overview. Journal of Nutritional Science 5. https://doi.org/10.1017/jns.2016.41
Park S, Choi MJ, Lee JY, Kim JK, Ha SH, Lim SH (2016). Molecular and biochemical analysis of two rice flavonoid 3’-hydroxylase to evaluate their roles in flavonoid biosynthesis in rice grain. International Journal of Molecular Sciences 17:1549. https://doi.org/10.3390/ijms17091549
Ren T, Zheng P, Zhang K, Liao J, Xiong F, Shen Q, Ma Y, Fang W, Zhu X (2021). Effects of GABA on the polyphenol accumulation and antioxidant activities in tea plants (Camellia sinensis L.) under heat-stress conditions. Plant Physiology and Biochemistry 159:363-371. https://doi.org/10.1016/j.plaphy.2021.01.003
Schrago C, Aguiar B, Mello B (2018). Comparative evaluation of maximum parsimony and Bayesian phylogenetic reconstruction using empirical morphological data. Journal of Evolutionary Biology 31(10):1477-1484. https://doi.org/10.1111/jeb.13344
Sharma M, Chai C, Morohashi K, Grotewold E, Snook ME, Chopra S (2012). Expression of flavonoid 3’-hydroxylase is controlled by P1, the regulator of 3-deoxyflavonoid biosynthesis in maize. BMC Plant Biology 12:1-13. https://doi.org/10.1186/1471-2229-12-196
Ueyama Y, Suzuki KI, Fukuchi-Mizutani M, Fukui Y, Tanaka Y (2002). Molecular and biochemical characterization of torenia flavonoid 3’hydroxylase and flavone synthase II and modification of flower color by modulating the expression of these genes. Plant Science 163(2):253-263. https://doi.org/10.1016/S0168-9452(02)00098-5
Wang X, Xu Y, Chu C, Li H, Mi J, Wen Z, Zhang S, Wang Q, Quan S (2019). Effect of safflower yellow on early type II diabetic nephropathy: a systematic review and meta-analysis of randomized controlled trials. Journal of Pediatric Endocrinology & Metabolism 32(7):653. https://doi.org/10.1515/jpem-2018-0425
Wang YS, Xu YJ, Gao LP, Yu O, Wang XZ, He XJ, Jiang XL, Liu YJ, Xia T (2014). Functional analysis of Flavonoid 3’,5’-hydroxylase from Tea plant (Camellia sinensis): critical role in the accumulation of catechins. BMC Plant Biology 14(1):347. https://doi.org/10.1186/s12870-014-0347-7
Wei K, Chen H (2018). Global identification, structural analysis and expression characterization of cytochrome P450 monooxygenase superfamily in rice. BMC Genomics 19(1):35. https://doi.org/10.1186/s12864-017-4425-8
Wei K, Wang L, Zhang C, Wu L, Li H, Zhang F, Cheng H (2015). Transcriptome analysis reveals key flavonoid 3'-hydroxylase and flavonoid 3',5'-hydroxylase genes in affecting the ratio of dihydroxylated to trihydroxylated catechins in Camellia sinensis. PloS One 10(9):e0137925. https://doi.org/10.1371/journal.pone.0137925
Wu ZJ, Li XH, Liu ZW, Xu ZS, Zhuang J (2014). De novo assembly and transcriptome characterization: novel insights into catechins biosynthesis in Camellia sinensis. BMC Plant Biology 14(1):1-16. https://doi.org/10.1186/s12870-014-0277-4
Zelasko S, Palaria A, Das A (2013). Optimizations to achieve high-level expression of cytochrome P450 proteins using Escherichia coli expression systems. Protein Expression and Purification 92(1):77-87. https://doi.org/10.1016/j.pep.2013.07.017
Copyright (c) 2021 Notulae Botanicae Horti Agrobotanici Cluj-Napoca
This work is licensed under a Creative Commons Attribution 4.0 International License.
Open Access Journal:
The journal allows the author(s) to retain publishing rights without restriction. Users are allowed to read, download, copy, distribute, print, search, or link to the full texts of the articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author.