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CiT F3 Micro-ATX PC Gaming Case, MATX & ITX Mobo Support, Windowed Side Panel, Excellent Airflow, Space For 4 Cooling Fans, SD/TF Card Reader Inc, 2 x 120mm Red LED Fans Inc. | Black / Red Stripe

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Wang, Y., Liu, X. J., Chen, J. B., Cao, J. P., Li, X., Sun, C. D. (2022). Citrus flavonoids and their antioxidant evaluation. Crit. Rev. Food Sci. Nutr. 62, 3833–3854. doi:10.1080/10408398.2020.1870035 Liu, X. M., Hu, C. X., Liu, X. D., Riaz, M., Liu, Y., Dong, Z. H., et al. (2022). Effect of magnesium application on the fruit coloration and sugar accumulation of navel orange (Citrus sinensis osb.). Scientia Hortic. 304. doi:10.1016/j.scienta.2022.111282 Wang, F., Chen, L., Chen, H. P., Chen, S. W., Liu, Y. P. (2019). Analysis of flavonoid metabolites in citrus peels (Citrus reticulata “Dahongpao”) using UPLC-ESI-MS/MS. Molecules 24. doi:10.3390/molecules24152680 Correlation analysis and canonical correlation analysis between flavonoids content and expression of synthesis-related genes

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Citation: Xiong B, Li Q, Yao J, Liu Z, Yang X, Yu X, Li Y, Liao L, Wang X, Deng H, Zhang M, Sun G and Wang Z (2023) Widely targeted metabolomic profiling combined with transcriptome analysis sheds light on flavonoid biosynthesis in sweet orange 'Newhall' (C. sinensis) under magnesium stress. Front. Plant Sci. 14:1182284. doi: 10.3389/fpls.2023.1182284 Hauer-Jakli, M., Trankner, M. (2019). Critical leaf magnesium thresholds and the impact of magnesium on plant growth and photo-oxidative defense: a systematic review and meta-analysis from 70 years of research. Front. Plant Sci. 10. doi:10.3389/fpls.2019.00766The samples underwent a series of preparation steps, including immersion in liquid nitrogen, grinding to a fine powder, suspension in 70% methanol and centrifugation at 20,000 rpm for 20 minutes at 4°C. The supernatant was then filtered through a 0.22-μm nylon syringe filter before being subjected to UPLC-MS analysis. This involved the use of an Agilent SB-C18 column (2.1 × 100 mm, 1.8 μm), with mobile phase A consisting of water with 0.1% formic acid, while mobile phase B was a solution of 0.1% formic acid in acetonitrile. The elution procedure utilized a gradient of 0-95% B from 0-9 min, followed by a single minute at 95% B and further gradient steps of 95-5% B for 1 minute and 5% B from 11-14 minutes. The flow rate of the system was set at 0.35 mL/min, and the column temperature was held constant at 40°C, with an injection volume of 4 μL. Multiple reaction monitoring (MRM) mode was used to acquire data from the production scan, which was then analyzed with the Metabolites Database (METWARE database) for metabolite identification. Quantitative analysis of metabolites was then performed using Analyst 1.6.3 software, with further examination of the metabolic pathways of these compounds using the KEGG database ( http://www.kegg.jp). Transcriptome analysis CiT Pro Android X Gaming Cube White Case with 3 x 120mm Infinity ARGB Fans 1 x 6-Port Fan Hub Tempered Glass Front and Side Panels

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Du, H., Huang, Y., Tang, Y. (2010). Genetic and metabolic engineering of isoflavonoid biosynthesis. Appl. Microbiol. Biotechnol. 86, 1293–1312. doi:10.1007/s00253-010-2512-8 Matsia, S., Tsave, O., Hatzidimitriou, A., Salifoglou, A. (2022). Chromium flavonoid complexation in an antioxidant capacity role. Int. J. Mol. Sci. 23. doi:10.3390/ijms23137171 Flavonoids are a significant group of secondary polyphenolic metabolites with a chemical structure of 3-C (C6-C3-C6) ( Li etal., 2020b; Nabavi etal., 2020). These compounds are widely distributed throughout the plant kingdom, and more than 6,000 distinct flavonoids have been identified to date ( Lepiniec etal., 2006; Ferrer etal., 2008). The pathways involved in flavonoid metabolism have been extensively studied in model plants ( Deng and Lu, 2017; Tohge etal., 2017). The flavonoid biosynthesis process begins with the primary glucose produced through photosynthesis, which is then converted into phenylalanine through glycolysis, pentose phosphate, and shikimic acid pathways. Phenylalanine enters the phenylpropane metabolic pathway through the action of phenylalanine ammonylase (PAL). In this pathway, phenylalanine is converted into p-coumaryl CoA through a series of reactions catalyzed by PAL, cinnamate 4-hydroxylase (C4H), and 4-coumaroyl CoA ligase (4CL) ( Forkmann and Martens, 2001; Du etal., 2010; Saito etal., 2013). The flavonoid biosynthesis pathway is initiated by condensation of one molecule of 4-coumaroyl-CoA with three molecules of malonyl CoA by chalcone synthase (CHS) enzyme. Subsequently, chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS) lead to the synthesis of anthocyanidin pigments. Flavone synthase (FNS) and isoflavone synthase (IFS) produce flavones and isoflavones, respectively. Flavonol synthase (FLS) catalyzes dihydroflavonols to flavonols, while leucoanthocyanidin reductase (LAR) and anthocyanidin reductase (ANR) synthesize cis- or trans-flavan-3-ols, which are precursors of proanthocyanidin (PA) polymers ( Owens etal., 2008; Li etal., 2020a). Regulation of genes involved in flavonoid biosynthesis is controlled through specific mechanisms that vary depending on the species or tissue involved ( Zhang etal., 2022). The accumulation of flavonoids is mainly controlled by structural biosynthetic genes, regulatory MYB transcription factors (TFs) and the MYB-bHLH-WD40 (MBW) complex ( Appelhagen etal., 2011; Hichri etal., 2011; Song etal., 2019; Yang etal., 2023). These factors are responsible for regulating the expression of genes involved in flavonoid biosynthesis, leading to their accumulation. CiT Vento White Micro-ATX PC Gaming Case with 4 x 120mm ARGB Fans Included 1 x 6-Port Fan Hub Tempered Glass Side Panel Li, Y., Chen, Q. Y., Xie, X. D., Cai, Y., Li, J. F., Feng, Y. L., et al. (2020b). Integrated metabolomics and transcriptomics analyses reveal the molecular mechanisms underlying the accumulation of anthocyanins and other flavonoids in cowpea pod (Vigna unguiculata l.). J. Agric. Food Chem. 68, 9260–9275. doi:10.1021/acs.jafc.0c01851Ahmed, O. M., Hassan, M. A., Abdel-Twab, S. M., Azeem, M. N. A. (2017). Navel orange peel hydroethanolic extract, naringin and naringenin have anti-diabetic potentials in type 2 diabetic rats. Biomed. Pharmacother. 94, 197–205. doi:10.1016/j.biopha.2017.07.094 Singh, B., Singh, J. P., Kaur, A., Singh, N. (2020). Phenolic composition, antioxidant potential and health benefits of citrus peel. Food Res. Int. 132, 109114. doi:10.1016/j.foodres.2020.109114 CiT Saturn White Micro-ATX PC Gaming Case with 4 x 120mm Infinity ARGB Fans Included 1 x 4-Port Fan Hub Tempered Glass Side Panel

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Deng, Y. X., Lu, S. F. (2017). Biosynthesis and regulation of phenylpropanoids in plants. Crit. Rev. Plant Sci. 36, 257–290. doi:10.1080/07352689.2017.1402852 Ramakrishna, A., Ravishankar, G. A. (2011). Influence of abiotic stress signals on secondary metabolites in plants. Plant Signal Behav. 6, 1720–1731. doi:10.4161/psb.6.11.17613

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Zhang, B., Yang, H. J., Qu, D., Zhu, Z. Z., Yang, Y. Z., Zhao, Z. Y. (2022). The MdBBX22-miR858-MdMYB9/11/12 module regulates proanthocyanidin biosynthesis in apple peel. Plant Biotechnol. J. 20, 1683–1700. doi:10.1111/pbi.13839 Li, P., Yao, X., Zhou, Q. Q., Meng, X., Zhou, T., Gu, Q. (2022a). Citrus peel flavonoid extracts: health-beneficial bioactivities and regulation of intestinal microecology in vitro. Front. Nutr. 9. doi:10.3389/fnut.2022.888745 To analyze the relative levels of 740 flavonoids and the expression levels of differentially expressed genes in the transcriptome, principal component analysis (PCA) was performed using the default parameters of the Metware data processing platform ( https://www.metware.cn/). Canonical correlation analysis (CCA) was conducted to explore the potential correlation between flavonoid metabolite levels and synthetic genes through metabolomic and transcriptomic data. Specifically, 21 flavonoid content levels and the expression levels of 41 hub genes within the flavonoid synthesis pathway of MS and MD peels at various stages were imported into Canoco 5.0 for CCA analysis, utilizing the default parameters. Quantitative real-time PCR analysis Zhou, H., Lin-Wang, K., Wang, H., Gu, C., Dare, A. P., Espley, R. V., et al. (2015). Molecular genetics of blood-fleshed peach reveals activation of anthocyanin biosynthesis by NAC transcription factors. Plant J. 82, 105–121. doi:10.1111/tpj.12792

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